PP4 - Displayed on Wednesday, September 12

PP4 - Displayed on Wednesday, September 12

Gynecopathology PP4-1 DIFFERENTIAL DIAGNOSIS PROBLEMS IN CHORIOCARCINOMA Eliza Gramada1, Sabina Zurac1, Alina Georgescu1, Razvan Andrei1, Florica Staniceanu1, Carmen Ardeleanu2, Bogdan Mastalier3 1 Colentina University Hospital, Department of Pathology, Bucharest, Romania 2 Victor Babes Institute, Bucharest, Romania 3 Colentina University Hospital, Department of Surgery, Bucharest, Romania

Background: Choriocarcinoma is a relative rare malignant tumor of gestational trophoblastic disease. Most cases present myometrial invasion, but rarely locoregional metastases (vulva, vagina, broad ligament) or in other organs (lung, brain, spleen, gastrointestinal tract, liver) can be found. Material and method: We present a case of a 41-years-old woman who was admitted in emergency in Universitary Colentina Hospital for massive vaginal bleeding; a tumoral abdominal mass was present at clinical examination. For these reasons the patient underwent a surgical intervention (total hysterectomy and bilateral oophorosalpingectomy). Results: Gross appearance: the uterus is much larger (16/12/7,5cm), fluctuent at palpation; the uterine cavity was occupied by numerous cysts measuring 0,3 to 1 cm across with serous/serohematic content forming an adherent mass to the endometrial mucosa; there are necrotic areas especially in fundic zone. Microscopical examination revealed rare hydropic villi and a tumoral proliferation composed of trophoblastic cells with insular/ cordonal pattern invading the myometrium. Tumoral cells have a marked cellular and nuclear pleomorphism, with frequent atypical mitoses. Occasionally, in periphery of the tumor, giant multinucleated tumoral cells with marked cytonuclear atypia could be identified. This atypical aspect of trophoblastic cells invited us to a large debate of the differential diagnosis with invasive mole caused by the presence in a limited aria of the hidropic villi. Hormonal tests have been performed (beta HCG >40 000 mU/ml) and immunohistochemical tests: positive reaction for betaHCG in hydropic villi and in trophoblastic elements; p53 was positive 10-15% and Ki67 was positive 3-4% in trophoblastic elements, all of them confirmed the histopathological result. Conclusion: We presented this case both for the rarity of gestational trophoblastic disease and for the difficulty to sustain the histopathological diagnosis. We want to evidentiate the utmost importance of the periodical examinations in pregnancy.

PP4-2 EXPRESSION OF AURORA KINASES A AND B IN OVARIAN CARCINOMA: CORRELATION WITH CLINICOPATHOLOGICAL FEATURES Marta Mendiola1, Jorge Barriuso2, Adrian Mariño-Enriquez1, Gines Hernandez-Cortes3, Elia Perez4, Aurora Dominguez-Caceres1, Andres Redondo2, Juan Angel Fresno-Vara2, Asuncion Suarez1, Jose Palacios5, David Hardisson1 1 Dpt. of Pathology, Hospital Universitario La Paz, UAM, Madrid, Spain 2 Dpt. of Medical Oncology, Hospital Universitario La Paz, UAM, Madrid, Spain 3 Dpt. of Gynaecology, Hospital Universitario La Paz, UAM, Madrid, Spain 4 Section of Biostatistics, Hospital Universitario La Paz, UAM, Madrid, Spain 5 Dpt. of Pathology, Hospital Universitario Virgen del Rocio, Sevilla , Spain

BACKGROUND: Ovarian carcinoma is the most important cause of gynaecological cancer-related mortality in the western world with a poor 5-year survival. Combination platinum-paclitaxel chemotherapy has become a standard first line treatment. Aurora kinases are involved in mitosis and cell division process. Overexpression of Aurora kinases, particularly Aurora-A, has been reported in many cancers. The aim of this study was to evaluate the expression of Aurora kinases A and B in ovarian carcinomas and to correlate their expression with clinicopathological features. MATERIALS AND METHODS: The study was conducted on 68 ovarian carcinomas treated by surgery followed by paclitaxel plus carboplatin-based chemotherapy. A complete response to chemotherapy was observed in 66.2% of cases. Median follow-up of survivors (38.2%) was 33 months. Antibodies against Aurora-A (Novocastra, 1:100), Aurora-B (Bethyl, 1:50), estrogen receptor (ER) (Novocastra, 1:100), progesterone receptor (PR) (Dako, 1:100), p53 (Novocastra, 1:100) and Ki67 (Dako, 1:100) were applied using the Envision (Dako) method. The immunohistochemical expression of Aurora-A and –B was evaluated as present or absent, regardless of intensity and percentage of cells that exhibit immunostaining. Additionally, we analyzed the amplification of Aurora-A gene by FISH. RESULTS: Overall, 58.8% and 92.1% of ovarian carcinomas showed overexpression of Aurora-A and –B, respectively. Expression of Aurora-A and –B did not correlate with FIGO stage. However, disease free survival (DFS) was significantly higher among patients with Aurora-A protein expression (P=0.02). A clear trend for longer DFS was also observed in Aurora-B expressing carcinomas (p=0.06). Additionally, a trend for positive correlation between Aurora-A expression and complete response to chemotherapy was also observed (p=0.09). Amplification of Aurora-A was found in 26.7% of cases examined, and was not associated with overexpression of Aurora-A by immunohistochemistry. There was no significant association of Aurora-A amplification and the recorded clinicopathological variables. Expression of ER, PR, p53 and Ki67 did not show association with clinico-pathological variables. CONCLUSION: Aurora-A expression seems to have a prognostic value in patients with ovarian cancer treated with a platinum-taxol based regimen. The present results are consistent with a role of Aurora kinases as a determinant of chemosensitivity of ovarian carcinomas. However, these results should be confirmed in a larger series of ovarian carcinomas. Grant (MEC) SAF2004-0825-C02-02

PP4-3 A CASE REPORT: EXTENSIVE IN SITU SQUAMOUS CELL CARCINOMA OF THE ENDOMETRIUM AS SUPERFICIAL EXTENSION OF INVASIVE CERVICAL CARCINOMA Cigdem Himmetoglu1, Gamze Mocan Kuzey1, Ali Ayhan2 1 Hacettepe University Faculty of Medicine Department of Pathology, Turkey 2 Hacettepe University Faculty of Medicine Department of Gynecology and Obstetrics, Turkey

Background: We report a case of 67-year-old woman with cervical squamous cell carcinoma (SCC) with extensive in situ squamous cell carcinoma of endometrium.SCC is the most common tumor of the female genital tract.However primary SCC of endometrium is extremely rare and its diagnosis requires absence of cervical malignancy. It is commonly encountered in postmenopausal women, where chronic pyometra or endometritis and long-lasting use of an intrauterine device are proposed etiologic factors.Ichthyosis uteri is a rare condition describing epidermalisation of the uterine cavity.It was described as an endometrial response to iatrogenically-introduced caustic substances and with a variety of inflammatory conditions of the endometrium. Case:A multiparous woman with new onset

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vaginal bleeding administered to our Gynecology and Obstetrics Department’s outpatient clinic.Physical examination revealed a massively enlarged, barrel-shaped cervix.Following a biopsy, she underwent a type III radical hysterectomy, bilateral salpingo-oophorectomy and pelvic-paraaortic lymph node sampling.Macroscopic examination revealed a 4 cm × 4 cm exophytic mass obliterating the endocervix and extending into the lower uterine segment.Sectioning revealed stromal invasion to almost 100% of the cervical wall.The uterus has become “pyometria” and uterine wall was extremely thinned.The endometrium was flat and glistening but was otherwise unremarkable.Microscopically,cervical neoplasm was a moderately differentiated,large cell keratinizing SCC.Extensive sampling of the endometrial wall revealed complete epidermalisation and areas where cells displayed high grade squamous intraepithelial lesion or carcinoma in situ.There was no evidence of extrauterine disease, parametrium was free of disease, 2 lymph nodes were metastatic.The patient received chemotherapy, now alive without diseases. Conclusion: Our case is a cervical SCC associated with extensive ichthyosis uteri-like changes of the endometrium that, additionally,had superimposed carcinoma in situ. These coincident findings may be explained that a SCC in the cervix and the associated HPV extended proximally,colonizing a pre-existing ichthyosis uteri.The second potential explanation is that within a background of extensive ichthyosis uteri, a SCC developed in the lower uterine segment.Neoplastic potential of ichthyosis uteri is largely unknown and theoretically longstanding,mature and even keratinizing squamous epithelium in the endometrium would be subject to the same potential changes in other visceral sites.

PP4-4 OVARIAN MUCINOUS CYSTADENOCARCINOMA WITH MURAL NODULE R. Nilgun Demirbag1, Gulbin Oran1, Pınar Cilesiz Goksedef2, Gonca Kolukfaki Batmaz2 1 Haseki Training and Research Hospital, Department of Pathology, Turkey 2 Haseki Training and Research Hospital, Department of Obstetric and Gynecology, Turkey

Solid mural nodule within a mucinous cystic ovarian tumor is extremely rare. Sarcoma-like mural nodules are not just limited to mucinous tumor. They can also seen in squamous cell carcinoma, cystic teratoma and granulosa cell tumor. A 78 year-old woman presented with abdominal pain caused by a mass. USG detected a 199x150 mm cystic mass.The mass contained a solid area of 73x47 mm and had a rich neovascularisation. The serum levels of CA125, CA15.3, AFP, CEA were normal and CA 19.9 was slighly eleveted at 50,9 U/ml. Grossly the ovarian tumor measured 17x16x11 cm. The outer surface was smooth and gray-white in colour. The cut surface showed a hemorrhagic mural nodule of 8x8x5 cm. Microscopically, tumor showed features of a malignant mucinous epitelial tumor. Mural nodule composed of round or ovoid cells with pleomorfic nuclei and prominent nucleoli. There was clear nuclear atypia and mitotic figures were inconspicous. There were scattered multinucleated giants cells with plemorfic hyperchromatic nuclei. Lymphovasculer and stromal invasion were present around the tumoral focus. Immunohistochemically the atypical cells in the mural nodule were negative with SMA and desmin. They expressed vimentin and multinuclear giant cell were CD68 positive. There were scattered CK positive atypical glands and isolated cells. In the literature, rare cases of ovarian mucinous tumors have been described that contain foci of sarcoma-like nodules. This case report describes the pathologic features of this rare entity with review of the literature.

PP4-5 DISTRIBUTION OF OVARIAN TUMORS IN ZONGULDAK KARAELMAS UNIVERSITY MEDICAL FACULTY HOSPITAL BETWEEN 2001-2007 YEARS Figen Barut1, Gurkan Kertis1, Aykut Barut2, Sibel Bektas1, Banu Dogan Gun1, Burak Bahadir1, Gamze Yurdakan1, Sukru Oguz Ozdamar1 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Zonguldak Karaelmas University, Faculty of Medicine, Department of Obstetric & Gynecology, Zonguldak, Turkey

Introductıon: Ovarian tumors are one of the common form of neoplasia in women. It is the fifth leading cause of cancer mortality in women. There are numerous types of ovarian tumors, both benign and malignant. Majority are benign. The purpose of this study is to evaluate the epidemiology of ovarian cancer in Zonguldak and surrounding cities, retrospectively. Methods: Between May 2001 and March 2007, 769 ovarian biopsies from 494 cases which undergo operation for variable cause are reviewed in Karaelmas University Medical Faculty Pathology Department. Tumors and other pathologic changes were classified. Results: Patients ages are between 10 days to 85 years and average is 45.46 (SD±13,45) years. Ovarian tumor’s ratio is (n: 123) 24,9% in all cases. Distribution of this 123 ovarian tumor: 90 (%73,2) are benign, 9 (%7,3) are borderline and 24 (%19,5) are malignant. 68 of the cases (55,3%) are tumors of surface epithelial origin of the ovary {26 (38,2%) are benign serous, 18 (26,5%) benign mucinous, 2 (2,9%) benign brenner, 7 (10,3%) borderline serous, 2 (2,9%) borderline mucinous, 9 (13,2%) malignant serous, 1 (1,5%) transitional cell carcinoma, 1 (1,5%) endometrioid carcinoma, 1 (1,5%) clear cell carcinoma, 1 (1,5%) malignant mixt mullerian tumor}; 34 of the cases (27,7%) are germ cell tumors {33 (97,1%) mature cystic teratoma, 1 (2,9%) teratoma with malignant transformation}; 10 of the cases (8,1%) are sex-cord stromal tumors {1 (10%) adult granulosa cell tumor, 1 (10%) juvenil granulosa cell tumor, 4 (40%) fibroma, 2 (20%) fibrothecoma, 1 (10%) sclerosing stromal tumor, 1 (10%) sex-cord stromal tumor of not other specified}; one of the cases (0,8%) is vascular tumor (cavernous hemangioma); one of the cases (0,8%) is undifferantiated carcinoma and 9 are (7,3%) metastatic tumors. Conclusion: Among cancers of the female genital tract, the incidence of ovarian cancer ranks below only carcinoma of the cervix and the endometrium. Ovarian cancer mortality takes the first place when compared with all other gynecological malignancies. In our study cystadenoma and cystadenocarcinoma are the most frequent benign and malignant tumors respectively and most of them are serous. When compared with the literature, tumors of surface epithelial origin of the ovary are less frequent, but metastatic tumors to ovary are more frequent in both whole and malignant groups. Germ cell tumor’s ratio is higher according to literature. We also have cavernous hemangioma which is an extremely rare tumor of ovary. PP4-6 CLINICO-MORPHOLOGICAL PECULIARITIES OF SEPTICEMIA IN LABOR AS A SEPARATE NOSOLOGICAL FORM AMONG THE CAUSES OF MATERNAL DEATH Andrey Milovanov1, Alexander Matsionis2, Nina Mikhanoshina2 1 State Research Institute of Human Morphology of the Russian Academy of Sciences, Russia 2 Rostov Regional Institute of Pathology, Russia

Background -Pyoseptic diseases account for 15% of the main causes of maternal death and are usually classified as postnatal sepsis Methods - We studied 15 autopsies of puerperas who died over a 10 year period (1996 - 2006) from pyoseptic diseases in the Rostov Province, and we analyzed their afterbirths and partum histories. Microscopic examination of uteri, spleens and lymph nodes, as well as fetal membranes and placentas, was

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performed by standard methods of light microscopy, with hemotoxillin and eosin staining; PAS reaction was also used, and immunohistological verification of cells from the inflammatory infiltration in the above organs. Results - Microscopic and immunohistochemical examination of the uterus showed, in 50% of cases, shallow invasion of the cytotrophoblast cells in the placental bed, incomplete gestation restructuring of the endometrial segments of the uteroplacental artery; in 60% of cases we found focal, low-grade infiltration with granulocytes, T-lymphocytes, and macrophages; myometry revealed, in 60% of cases, focal low-grade, mostly perivascular, infiltration with granulocytes, T-lymphocytes, and macrophages. The inflammatory infiltrate in the placental fetal membranes was represented by a great number of granulocytes, plasmacytes, macrophages, and rare T-lymphocytes. The inflammatory infiltrate contained no B-lymphocytes. In 73% of cases we observed effacement of the spleen pattern, dysfunction of the humoral and cell protection - depletion of the lymphoid tissue and its replacement with granulocytes and macrophages; the lymph nodes in 86% cases also showed effacement of the pattern, with only fragments of the lymphoid follicles preserved, while the parafollicular zone was characterized by depletion of T-lymphocytes and diffuse infiltration with granulocytes and macrophages. Conclusion - Early antenatal infection of the afterbirth, which is these cases is the entrance gate for penetration of bacterial, fungous, and viral agents into the mother's blood flow. We found a relative intactness of the uterus, because the revealed inflammatory changes in the basal endometry and myometry should be regarded as normal inflow of leucocytes and lymphocytes to the area affected by necrosis and rejection of the decidual tissue and remains of the fetal membranes. The rapidity of development of the clinical manifestations of septicemia in labor, and of the infection and toxic shock, leading to death of the puerperas within the first days of the postpartum period, is accounted for by the apparent deterioration of the immune system function. PP4-7 HIGH-GRADE ESS ARISING FROM OVARIAN ENDOMETRIOSIS, WITH ASSOCIATED FOCI OF ENDOMETRIOID ADENOCARCINOMA: REPORT OF A CASE. Cristina Manieli, Giuseppina Parodo, Sara Pillai, Gavino Faa Department of Cytomorphology, Section of Pathology, University of Cagliari, Italy

Background: Endometriosis malignant transformation is a well documented phenomenon that occurs most commonly in the ovaries, frequently associated with endometrioid or with clear cell adenocarcinomas. Endometrioid stromal sarcoma (ESS) is a sarcomatous tumor characterized by a diffuse proliferation of neoplastic cells similar to the normal stromal cells present in the proliferative endometrium. Among these, ESS arising in endometriosis of the ovary is an extremely rare tumor. Here we present, for the first time, a case of an high-grade ESS arising from ovarian endometriosis associated with endometrioid adenocarcinoma foci. A 51-years-old white woman, with no significant gynecological history, presented with increasing abdominal distension, and weight gain. After an exploratory laparotomy a 10 cm right ovarian mass with neoplastic ascitis and multiple tumoral nodules in the omentum and in the peritoneum was found. Given the advanced stage, debulking surgery was performed. No alteration of the uterine corpus were revealed under clinical, ultrasonographic and laparoscopic examinations. The patient died after 1 month for the diffuse metastatic disease. Method Immunohistochemical studies were performed by indirect staining methods using antibody against cytocheratin 7, 20, 8-18, vimentin, epithelial membrane antigen (EMA), inhibin, calretinin, S100 protein, HMB45, smooth muscle actin, desmin, estrogen receptor (ER), progesterone receptor (PR), NSE, cromogranin, sinaptofisin, CD99, CD45, CD30, Ki-67 (Mib-1),

CD10. Results The ESS tumor nodules were composed mostly by sheets or cords of closely packed round or oval uniform cells, with pale and scanty cytoplasm and intersected by dense fibrous band. Some of these neoplastic cells occasionally were found around numerous small thick-walled blood vessels. More than 10 MF per 10 HPF were counted. Extensive necrosis and hemorrhage and vascular and perineural invasion were present in the tumor. Endometriosis, identified in the cystic areas, in some areas was associated with endometrioid adenocarcinoma foci (G2, nuclear grade 3). ESS tumor cells immunohistochemical stain was positive for vimentin, CD10 and focally for EMA and around 80% of nuclei were Ki-67 (Mib-1) positives. All the other immunohistochemical markers performed were negative. In the multiple foci of of endometrioid adenocarcinoma the neoplastic glands resulted diffusely positive for EMA and CK7, and negative for CD10. Conclusion: According to our knowledge this is the first report of a high-grade ESS associated with concomitant endometrioid adenocarcinoma foci. PP4-8 MULTIPLE OXYPHILIC TYPE ADENOMATOID TUMORS OF THE UTERUS COEXISTING WITH ENDOMETRIAL ADENOCARCINOMA IN A RENAL TRANSPLANT RECIPIENT. Antigoni Karakosta, Maria Karagiannis, Rosaria Mennonna, Eleni Sotiriou, Panagiota Pantoula, Sofia Xyristaki, Aikaterini Parasi Pathology Department, General Hospital of Nikaia “Agios Panteleimon”, Piraeus, Greece

Backround: Uterus is the most common site of occurrence of adenomatoid tumors in the female genital tract. Multifocal cases, as well as oxyphilic variants, have rarely been described. To our knowledge, this is the second case, coexisting with endometrial adenocarcinoma of the uterus and the fourth case, in a renal transplant recipient. Case: This is a report of the coexistence of a multifocal uterine adenomatoid tumor with a poorly differentiated endometrioid adenocarcinoma, FIGO stage IB. Both were incidental findings in a 56 years old woman, undergoing hysterectomy because of large leiomyomas. Two of the adenomatoid tumors, found macroscopically, had gelatinous appearance while the third was found microscopically. All consisted of large oxyphilic epithelioid type cells, with a solid or trabecular pattern resembling leiomyoblastoma. Slit–like and adenoid pattern was rarely found. Immunohistochemically, the tumor cells stained for Vimentin, Pankeratin, Calretinin, Keratin 5/6 and HBME-1, while no immunoreactivity for Actin, Desmin. H-Caldesmon, CD34 and CD31 was found. The Ki67 index was low. Conclusion: Uterine adenomatoid tumors are rare tumors, which may cause differential diagnostic problems, but the knowledge of their morphological spectrum and the characteristic immunoprofile allows their diagnosis. Very interesting features of our case are the multicentricity and the oncocytic appearance of the tumor, as well as the coexistence with a poorly differentiated carcinoma, in an immunocompromissed status of a renal transplant recipient. PP4-9 PRIMARY VAGINAL MUCINOUS ADENOCARCINOMA ARISING ON A PREEXISTING ADENOMA. A RARE CASE REPORT. Aikaterini Apostolaki, Eleni Ieremia, Efthimios Koniaris, Maria Gazalidou, Maria Mpiteli, Apostolikas Nikiforos Pathology Department, Anticancer Oncologic Hospital of Athens “St. Savvas”, Athens, Greece

Background: Primary vaginal mucinous adenocarcinomas (PVMA) are rare neoplasms that develop in patients with in utero exposure to diethylstilbestrol (DES). We report a case of PVMA of intestinal type arising on a preexisting adenoma, in a patient without intra-uterus DES exposure. Awareness of this lesion is important as it must be distinguished from metastatic

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adenocarcinomas especially from colon or endocervix. Method: A 13mm polypoid mass was found on the posterior vaginal wall of a 52-year-old menopausal woman during routine gynecological check up. The patient did not have any symptoms of vaginal bleeding, mucus secretion nor a history of diethylstilbestrol exposure. Complete excision of the polypoid mass was performed and the histological examination showed adenocarcinoma of intestinal type arising on a tubular adenoma. To preclude possible metastasis from the gastrointestinal tract, sigmoidoscopy, colonoscopy and gastroscopy was ordered showing no primary gastrointestinal lesions. Results: Grossly, the specimen was a lobulated, circumscribed, brown-colored solid lesion measuring 13mm. Histologically, it was a tubular adenoma of medium degree of dysplasia with glands made of columnar and goblet cells without any Paneth and neuroendocrine differentiation. The adenoma focally transformed to areas with higher density of glands, placed back-to-back, with multistratification of nuclei, loss of cell polarization and cribriform architectural arrangement, similar to in situ intestinal adenocarcinoma arising on an adenoma. The adenocarcinomatous area was showing early focal invasion of the underlying stroma with accompanying chronic inflammatory infiltration and slight desmoplasia. Dysplastic glandular epithelium was replacing the normal vaginal non-keratinized stratified squamous epithelium in adjacent sites. The immunohistochemical profile of the lesion revealed expression of CK 20 and CEA and negativity in CK 7. Alcian blue positivity was indicative of intestinal type mucin. Conclusion: PVMA is a rare tumor of the lower female genital tract and its occurrence is higher among women who have been exposed to DES during intra-uterus life. However, cases of no such association to DES have also been reported. The differential diagnosis involves metastatic mucinous adenocarcinoma of the endocervix and of the intestines. In the first case, the adenocarcinoma cells contain sulfomucin, a mucin specific for adenocarcinoma of the endocervical type. In the second case, the mucin is the intestinal type o-acetylated sialomucin. PP4-10 HIGH CELLULAR LEIOMYOMA WITH EXTENSIVE CYSTIC DEGENERATION MIMICKING LOW GRADE ENDOMETRIAL STROMAL SARCOMA. A CASE REPORT WITH PECULIAR MACROSCOPIC APPEARANCE Apostolaki Aikaterini, Efthimios Koniaris, Eleni Ieremia, Antigoni Papathanasaki, Maria Sevastiadou, Nikiforos Apostolikas Pathology Department, Anticancer Oncological Hospital ‘St. Savvas’, Athens, Greece

Background: High Cellular Leiomyoma (HCL) is a subtype of Cellular Leiomyoma (CL) with accentuated cellularity composed of small, round or spindle cells lacking moderate to severe cytological atypia and necrosis. HCL is often misdiagnosed as Endometrial Stromal Tumor (EST). We present a case of HCL in the form of an intramural, solitary, well-defined myometrial cyst. Method: A 38-year-old woman presented with a pelvic mass, causing urinary urgency and stagonoid hemorrhage. On palpation, and ultrasonographically, an enlarged cystic uterine mass was observed, displacing the urinary bladder and slightly obstructing the ureters causing dilation of the pyelocalyceal system. On CT and MRI scan no other notable lesions were discovered. Total hysterectomy was performed with bilateral salpingectomy. Results: Grossly, the uterus measured 13.5x11x10.5cm and the left and right salpinx measured 3.5 and 3.7cm respectively. On dissection, a voluminous cystic mass was observed in the ventral myometrial wall measuring 9cm in greatest diameter and filled with a yellowish serous liquid content. On cross section, the mass was whitish-yellowish-to-tan, nodular-appearing, with extensive cystic degeneration and soft in consistency. A small leiomyoma measuring 1.5cm was also found. The endometrial surface was not affected. Histologically,

the tumor was characterized by a monotonous appearing hypercellular mass with slightly irregular peripheral borders. It was composed of relatively uniform cells showing no or minimal cytological atypia, having round or ovoid nuclei with finely granular and evenly dispersed chromatin, small inconspicuous nucleoli and scanty cytoplasm. At foci, typical smooth muscle bundles and few thick walled vessels were encountered. Furthermore, cystic changes were also spotted among the stromal neoplastic cells. Low mitotic activity (infrequent mitotic figures, <5/10HPF) was evident. The margins of the tumor were relatively well-delineated with focal finger-like irregularities of 3mm and isolated neoplastic cells infiltrating the adjacent myometrium. Necrotic foci and invasion of the lymphatic and vascular channels were not observed. Conclusion: Differential diagnosis includes Endometrial Stromal Nodule (ESN) and low grade Endometrial Stromal Sarcoma (ESS). Although morphologically the neoplasm is consisted with low grade ESS and despite the slightly irregular peripheral borders of the neoplasm, HCL is strongly considered. Our case was positive in vimentin/ER/PR/desmin/SMA and negative in CD10, the latter typically staining endometrial stromal tumors. PP4-11 DIVERSITY OF MORPHOLOGY, PROGNOSIS AND CLINICS OF ENDOMETRIAL CARCINOMA Regina Kleina Riga Stradins University, Latvia

Endometrial carcinoma (EC) is the 3rd common cancer of female in Latvia with 30,8%o and there is tendency for increase. The aim of our research is to evaluate stage, variants, prognostic factors and immunohistochemical spectrum of EC. Materials and Methods. We have analyzed retrospective material of operated uterus from 60 cases with the characteristics of lymph nodes, tubes, ovaries and expression of estrogen, progesterone, CD 44 and synaptophysin markers. Coherence between clinical symptoms we analyzed by Speerman rank system, but correlations between morphological changes by diagrams. Results. 60% of EC were endometroid adenocarcinomas, 23%-villoglandular variant, 15%-with squamous differentiation, 2%-clear cell adenocarcinoma. Age of patients: 44-82 years. EC were of I to IV stage and myometrial invasion from 0, 1 to 2 cm. Clinical information showed uterine bleeding in 43%, obesity -5% of patients. Number of deliveries and abortion were variable. In 31, 6% of cases EC were proved in such precursor lesions as endometrial polypus and atypical hyperplasia. Mean age of menopause – 52 year. Late menopause was in 36% of patients. The size of the uterus in majority of cases was normal. Clinical diagnosis of benign process was changed to malignancies by pathologists in 14, 6 % of cases. Metastasis were proved only in 3, 3% of female. In 92% of patients lymph nodes were not exceeded for histological examination. In ovaries were different cysts. Expression of estrogen, progesterone markers were: negative in 23, 8 %, positive-57, 2 %, but in 19 % of cases estrogen was negative, but progesterone –positive. Conclusions.1.Diagram analyses showed the most aggressive invasion in myometrium by villoglandular variation of EC. 2. There were no statistically significant differences between different clinical data but we proved direct correlations between nononcological changes in ovaries with increased risk ofendometrial carcinoma at earlier age of patient. 3. Immunoreactivity of estrogen and progesterone receptors was more expressed in G1, G 2 of EC but lack of lymph nodes in our research group decreased the possibility of real prognosis.

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PP4-12 HOW OFTEN HSIL ACCOMPANIES LSIL: IS AGE IMPORTANT? Handan Cetiner1, Ayse Gurbuz1, Gozde Kir2, Ates Karateke1 1 Zeynep Kamil Hospital, Pathology Department, Turkey 2 Umraniye Hospital, Pathology Department, Turkey

BACKGROUND: High-grade squamous intraepithelial lesion (HSIL) has much greater potential for progressing than does low-grade intraepithelial lesion (LSIL). So it may be the important to identify the subset of women at higher probability of having “LSIL harbouring the HSIL” lesions in follow-up biopsy specimen whose cytologic specimens show atypical squamous cells of undetermined significance (ASC-US), atypical squamous cells; cannot exclude HSIL(ASC-H) and LSIL. METHOD: Sixty-five consecutive patients whose follow-up cervical biopsies were available and have ASC-US, ASC-H and LSIL lesions in their conventional smears were enrolled. We divided them in two groups: age 40 and under and over age 40. All follow-up biopsy results were evaluated and cases were excluded which the histologic examination didn’t show any squamous lesion. RESULTS: Six patients were excluded because of not showing any squamous lesion in their follow-up biopsy specimen.There were 39(66%) women age 40 and under and 20(34%) women over age 40 who have cytologic abnormalitiy as ASC-US, ASC-H and LSIL. In younger age group; 30(77%) women showed LSIL and 9(23%) women showed HSIL in their biopsy specimen. In over 40 group; there were 8(40%) LSIL and 12(60%) HSIL showing biopsy specimens. CONCLUSION: Compared to younger women with abnormal low-grade cervical cytology, women over 40 years of age, had higher rate of having ‘LSIL harbouring HSIL’ in their follow-up biopsy. Since current study has small sample group further studies involving large number of cases and detailed statistical analysis must be made. If these findings confirmed in future studies then it may be possible to say that HSIL lesion is more expectable in follow-up biopsy of women over 40 years of age with a diagnosis of ASC-US, ASC-H or LSIL in cervical cytology.

PP4-13 ENDOCERVICAL-LIKE MUCINOUS BORDERLINE TUMORS OF THE OVARY : A CLINICOPATHOLOGIC ANALYSIS OF 9 CASES Raoudha Doghri, Maha Driss, Samia Sassi, Imen Abbes, Karima Mrad, Ryme Dhouib, Naziha Ben Hamida, Khaled Ben Romdhnae Institut Salah Azaïz, Tunisia

Endocervical-like mucinous borderline tumors (EMBTs) are rare distinct entity of the ovary that seems to be under recognized. These tumors are characterized by a papillary architecture reminiscent of serous tumors but composed of endocervical-like mucinous epithelium. The latter may be admixed with other mullerian-type epithelium, including those of serous, endometrioid, squamous and eosinophilic epithelial cells. We have studied nine EMBTs including 2 cases with intraepithelial carcinoma and one case with stromal miroinvasion. The patients averaged 35.5 years of age. None tumor was bilateral. Endometriosis was identified in three cases and one patient was known to be pregnant at the time of diagnosis. All patients were stage I. Of the 5 patients which have a follow-up data, only one patient was complicated by recurrence in the conserved contra lateral ovary with a peritoneal implant, three years later. The present study describes our experience with endocervical type mucinous tumors. The frequency of occurrence, criteria for the diagnosis of intraepithelial carcinoma and microinvasion are discussed.

PP4-14 GENOMIC INTEGRATION OF ONCOGENIC HPV AND GAIN OF THE HUMAN TELOMERASE GENE TERC ARE STRONGLY ASSOCIATED EVENTS IN PROGRESSION OF UTERINE CERVICAL DYSPLASIA TO INVASIVE CANCER Anton Hopman1, Wendy Theelen1, Simon Herrington2, Larry Morrison3, Frenk Smedts4, Frans Ramaekers1, Ernst-Jan Speel1 1 Department of Molecular Cell Biology, University of Maastricht, Netherland 2 Bute Medical School, St. Andrews, UK 3 Vysis Inc, Downers Grove, IL, USA 4 Foundation of Collaborating Hospitals of Eastern Groningen, Winschoten, Netherland

Background: Recently proposed markers for progression of CIN II/III to cervical carcinoma include integration of HPV into the host genome, genomic instability and an increase in chromosome 3q copy number. In particular, the gene coding for the RNA component of telomerase (TERC) at 3q26 has been implicated as possible candidate gene. Since it is not known to date how these events are temporarily related during cervical carcinogenesis, the aim of the present study was to assess the correlation between TERC copy number and physical status of HPV in high grade solitary precursor lesions of the uterine cervix (CIN II/III, n=17), lesions associated with a microinvasive cervical carcinoma (CIN III&mCA, n=13), and advanced invasive cervical carcinoma (invCA, n=7). Method: Fluorescence in situ hybridization (FISH) was applied to assess the copy number of the TERC gene and HPV integration and to correlate TERC gain with HPV integration during progression in cervical neoplasia. The TERC gene probe was applied in a mixture together with probes for the centromeres of chromosomes 3 and 7, used to determine the ploidy of these lesions. Results: Our study revealed that CIN II/III lesions with episomal HPV frequently show tetrasomy for TERC. In these cases the TERC gene copy number parallels the DNA ploidy of the lesions. Furthermore, TERC was increasingly gained with progression of CIN II/III (3 of 17), via CIN III&mCA (7 of 13) to invCA (5 of 7). In the cases exhibiting gain of TERC the virus was predominantly integrated. The finding that the combination of gain of TERC (three copies) and genomic integration of HPV was seen in 8 out of 10 diploid lesions, indicates that these events occur prior to overall aneuploidization. Our data show that gain and aneusomy for the TERC gene is strongly correlated with viral integration, which in turn is associated with the progression of CIN III to mCA and invCA (p<0.001). Conclusion: Genomic integration of oncogenic HPV and gain of the human telomerase gene TERC are strongly associated events in progression of uterine cervical dysplasia to invasive cancer.

PP4-15 HPV DNA DETECTION AND TYPING AS AN AID IN THE CLASSIFICATION OF SECOND NEOPLASMS IN WOMEN WITH UTERINE CERVICAL CANCER August Vidal Bel1, Betlem Lloveras Rubio2, Jordi Casalots Casadó3, Mónica Olivera Sáez4, Jo Ellen Klaustermeier4, Josep Maria Piulats Rodríguez5, Enric Condom Mundó3 1 Servei d’Anatomia Patològica, IDIBELL, Hospital Universitari de Bellvitge. L'Hospitalet de Llobregat Barcelona, Spain 2 Laboratori de Papillomavirus, IDIBELL, Institut Català d’Oncologia L'Hospitalet de Llobregat Barcelona, Spain 3 Servei d’Anatomia Patològica, IDIBELL, Hospital Universitari de Bellvitge. L'Hospitalet de Llobregat Barcelona, Spain 4 Laboratori de Papillomavirus, IDIBELL, Institut Català d’Oncologia L'Hospitalet de Llobregat Barcelona, Spain 5 Servei d’Oncologia Mèdica, IDIBELL, Institut Català d’Oncologia L'Hospitalet de Llobregat Barcelona, Spain

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INTRODUCTION: The vast majority of uterine cervical carcinomas are human papillomavirus (HPV) related neoplasms. In women with a cervical cancer and a distant lesion the histologic distinction of metastatic cervical cancer versus another primary tumor can be difficult and has important clinical implications. OBJECTIVE: To study the presence of HPV DNA in primary cervical cancer and in second neoplasms in the same patient. MATERIAL AND METHODS: Four patients with squamous cell carcinoma (in bone, lung, lateral neck lymph node and pleural fluid) and two with adenocarcinoma (in ascitic fluid) and a past history of uterine cervical cancer (diagnosed 1 to 11 years before) were studied. HPV DNA detection was performed both in the cervical tumors and in the second neoplasms (formalin fixed, paraffin embedded tissues) using a PCR based technique with GP5+/6+ primers for detection of the HPV L1 gene followed by reverse line blotting hybridization for genotyping. RESULTS: In five patients DNA of the same HPV type was found in the cervical tumor and in the second neoplasm (HPV16 in 4 squamous cell carcinomas and HPV18 in one adenocarcinoma), thus allowing to classify the second tumors as metastatic. In the remaining patient HPV DNA was not detected in any of the tumors tested. CONCLUSIONS: In women with a past history of an HPV-related cancer and a second neoplasm detection and typing of HPV DNA is helpful in the differential diagnosis between metastases and a second unrelated malignancy. PP4-16 EFFECTIVENESS FOR EVALUATION OF LYMPHATIC INVASION IN ENDOMETRIOID ADENOCARCINOMA OF THE UTERINE CORPUS Yasuka Miyakuni, Yuki Yamada, Keiko Abe, Atsushi Arakawa, Hiroshi Sonoue, Toshiharu Matsumoto Juntendo University , Japan

Backgrounds: Lymph-vascular involvement is one of the important prognostic factors in the patients with carcinoma of the uterine cropus. Conventionally, it has been determined morphologically using hematoxylin and eosin sections. In addition to that, various immunohistochemical stains have been attempted in order to confirm the accuracy of diagnosis of lymph-vascular involvement with section stained H&E. Recently, D2-40 immunostaining was demonstrated as the best immunohistochemical marker for the endothelium of the lymph vessel. The aim of this study is to clarify the effectiveness of immunohistochemical staining with D2-40 in determination of lymphatic invasion. Methods: We investigated that 40 patients with adenocarcinoma confirmed to the uterine corpus, who were underwent radical hysterectomy. The stages of tumors were 26 cases in pT1b, 14 cases in pT1c, and 7 cases were positive node among 40 cases. From each material, one section containing the deepest invasive part was selected for examination and stained with H&E and D2-40 immunohistochemically. Lymphatic invasion was evaluated by either H&E staining or D2-40. The concordance of these results was assessed using the kappa statistic ( ). Additionally, the relationship between lymph vascular involvement and lymph node metastasis was examined by using Fisher’s exact test. Results: The evaluation of lymphatic invasion with H&E staining resulted in 24 positive cases (positivity of 60%) and 16 negative cases (negativity of 17.5%). The result with D2-40 immunostaining showed 7 positive cases (positivity of 17.5%) and 33 negative cases (negativity of 82.5%). The concordance between H&E and D2-40 was moderate (57.5%, =0.53). The negative cases in the evaluation with H&E staining were always negative in the evaluation with D2-40. As a result, lymph vascular involvement might have been over-estimated by H&E staining. The correlation between lymphatic invasion and lymph node metastasis showed statistical significance, in both results with H&E and D2-40 (p=0.011 and p=0.0001, respectively). Conclusion: Lymphatic invasion determined by D2-40 staining could be much more accurate predictive factor for lymph node metastasis.

PP4-17 ENDOMETRIAL CYTOLOGY IN DIAGNOSIS OF ENDOMETRIAL CARCINOMA Pricop Mihai1, Musca Simona2, Stolnicu Simona3, Maxim Razvan1 1 University of Medicine and Pharmacy “Gr. T. Popa” Iasi, Romania 2 Laboratory of Histopathology, Clinical Hospital of Obstetrics-Ginecology “Cuza Vod ” Iasi, Romania 3 Department of Pathology, University of Medicine Targu Mures, Romania

Cytodiagnosis of the precancerous lesions of the endometrium has remained unclear compared to that for cervical lesions. Endometrial cytology and biopsy have been performed for patients with atypical genital bleeding, who were over fifty, postmenopausal or nulliparous. The cytologic diagnosis of endometrial cancer using material obtained with the Inocurette endometrial sampler was assessed for 64 patients.The cytologic findings for benign and malignant samples are described and illustrated in detail. Relative to other endometrial sampling devices, the Inocurette is inexpensive and was easily used by the gynecologist and well tolerated by the patients, with no complications and minimal discomfort.

PP4-18 EVALUATION OF COMBINED BCL-2/MDM-2 IMMUNOHISTOCHEMICAL EXPRESSION AS A PROGNOSTIC FACTOR IN EARLY STAGES OF INVASIVE CERVICAL CARCINOMAS Irina Prodanova, Katerina Kubelka-Sabit, Neli Basheska Department of Histopathology and Clinical Cytology, Institute of Radiotherapy and Oncology, Medical Faculty, Skopje, Republic of Macedonia

BACKGROUND: The present study was designed to evaluate the immunohistochemical expression of apoptosis regulating proteins (bcl-2, mdm-2 and p53) in correlation with proliferation (Ki-67), human papillomavirus (HPV) infection and other histopathological and clinical parameters in early stage cervical carcinomas and the estimation of their prognostic significance. Special attention was given to combined bcl-2/mdm-2 immunophenotypes in predicting the recurrence of the disease. METHOD: The subject of this study was a series of 83 surgically treated patients with cervical carcinoma confined to the uterine cervix (pT1b1/1b2), who subsequently received complete radiotherapy. The presence of HPV DNA was determined by the conventional method of in situ hybridization (ISH) and catalyzed reporter deposition signal amplification ISH. The immunostaining was performed using avidin-biotin-peroxidase complex method and the expression of the biological markers was semiquantitatively evaluated as the percentage of immunostained cells. RESULTS: During the clinical follow-up (mean 120.7, range 4.4-181 months) a relapse was diagnosed in 9 (10.8%) patients and the expected 5-, 10- and 15- year disease-free survival was 92.7%, 90.8% and 86.6%, respectively. The results of the univariate analysis indicate that significant predictive indicators for recurrence are: lymphonodal status, maximal tumor diameter, depth of stromal invasion, histological type and HPV DNA presence and type. Immunohistochemical markers showed the following correlations: increased expression of Ki-67 (P=0.031) and bcl-2 negativity (P=0.047) correlated with poor disease-free survival, while mdm-2 positivity showed borderline significance (P=0.051) and p53 expression had no influence on disease-free survival. Additional evaluation of combined bcl-2/mdm-2 expression showed that cases with bcl-2+/mdm-2- and bcl-2-/mdm-2+ immunophenotype had better survival (P=0.048) compared to bcl-2+/mdm-2+ and bcl-2-/mdm-2- phenotype. In the multivariate analysis, histological type, HPV DNA presence and the expression of Ki-67 have been selected as

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the most significant independent prognostic parameters (P=0.0024). CONCLUSION: The evaluation of combined bcl-2/mdm-2 immunohistochemical expression provides more relevant information for the prediction of the recurrence of the disease than their individual expression. However, neither individual expression of bcl-2 and mdm-2 nor their combined immunohistochemical expressions are independent predictors of prognosis in early stages of invasive cervical carcinomas.

PP4-19 SCLEROSING STROMAL TUMOR OF THE OVARY WITH SEX CORD ELEMENTS Stephanie Vgenopoulou1, Athina Androulaki1, Maria Sotiropoulou2, Antonios Lagadas3 1 First Department of Pathology, University of Athens, Medical School, ‘Laiko’ Hospital, Greece 2 Department of Pathology, ‘Alexandra’ Hospital, Athens, Greece 3 Department of Gynecology, ‘Laiko’ Hospital, Athens, Greece

Screrosing stromal tumor of the ovary is a distinctive type of benign stromal tumor characterized by a cellular pseudolobular pattern. The tumor cell population is composed of fibroblasts and round cells separated by hypocellular, oedematous or collagenous tissue. In the recent literature there have been very rare reports of the coexistence of sex cord elements in, otherwise typical, sclerosing stromal tumors of the ovary. We present a case of a seventeen-year-old girl with a sclerosing stromal tumor in which sex cord elements were observed. Multiple histological sections were studied and immunohistochemistry was performed with a wide spectrum of antibodies. Morphological and immunohistochemical findings led to the diagnosis of sclerosing stromal tumor with sex cord elements. The tumor cells stained positive for desmin, vimentin and SMA. Sclerosing stromal tumors comprise a very distinct type of ovarian stromal tumors with very characteristic histology, which is presented in detail.

PP4-20 THE IMPORTANCE OF THE TUMOR SUPPRESSOR GENE DESIGNATED PTEN AND OF THE PROLIFERATION MARKERS Ki-67 AND PCNA IN THE EVALUATION OF THE MALIGNANCY POTENTIAL IN ENDOMETRIAL HYPERPLASIA Sajin Maria, Chefani Alina Elena, Lazaroiu Anca Mihaela, Simion George, Secara Diana, Carstoiu Monica Universitary Emergency Hospital Bucharest, Romania

Introduction: Endometrial Hyperplasia (increase in gland/stroma ratio, disorder number and shape of the glands like modifications of the glandular epithelium: exhibits loss of nuclear polarity, stratification, etc) is a lesion frequently associated with the uterine leiomyomas Purpose: The Study was realized at the Universitary Emergency Hospital Bucharest, Romania on 210 endometrium biopsies taken in the period 2004-2005 from women with ages between 45 and 55. Material and methods: Hematoxylin –eosin stained slides of endometrial formalin fixed, paraffin embedded tissue has emphasized in 105 cases the simple hyperplasia (SH), in 75 cases the complex hyperplasia (CH), in 11 cases the simple atypical hyperplasia (SAH) and in 23 cases the complex atypical hyperplasia (CAH). We performed the indirect tristadial ABC method of IHC for 3 antibodies: PTEN, Ki-67 and PCNA on formalin fixed embedded tissue taken by biopsies from 50 cases (8 SH, 8 CH, 34 SAH and CAH). Results: PTEN was focal positive for SAH, diffuse for CAH and for 1-2 cases of SH and CH. Ki-67 and PCNA were also very frequent in group SAH and CAH. Conclusion: PTEN, Ki-67 and PCNA take part in the process of endometrial carcinogenesis following probably molecular pathways and determine the malignity potential of atypical hyperplasia of endometrium.

PP4-21 EGFR AND HER2/NEU EXPRESSION IN ENDOMETRIAL CARCINOMA. CORRELATION WITH TYPE AND STAGE Maria Sotiropoulou, Irini Papaspyrou, Christos Eftychiades, Christos Evaggelakos, Pantelis Markoulis, Demetra Theocharidou, Vassilios Pavlou, Sofia Markaki Histopathology Department, Greece

BACKGROUND: Epidermal Growth Factor Receptor (EGFR - Her1) and Human epidermal growth factor type II (Her2/neu) are members of the subfamily of tyrosine kinase transmembrane receptors. The expression of EGFR and Her2/neu has been documented in a variety of malignant neoplasms of epithelial origin as breast, ovary and lung. The expression of these markers correlates with advanced stage, poor prognosis and metastasis. The status of these oncoproteins in endometrial carcinoma, however, is not entirely clear. Aim of this study was the evaluation of EGFR and Her2/neu in endometrial carcinoma (endometrioid and serous –type I and II ), the relation to other prognostic factors and the possible therapeutic implications. MATERIAL and METHODS: Forty one cases of endometrial carcinoma were selected, 22 endometrioid (type I) and 19 serous (type II). Formalin fixed-paraffin embedded tissues were immunohistostained with EGFR (clone 31G7, Zymed, CA, USA) and Her2/neu (clone CB11, Bio Genex, CA,USA). Over-expression of Her2/neu was defined as moderate or strong membranous staining ( 2+ staining) in more than 10% of the cells. EGFR’s expression was interpreted as positive (any membranous staining) or negative. Tumors were subdivided into two stage groups, 1 and 2. RESULTS: Patients ranged in age from 27-84 years, mean age 59,81 in endometrioid and 68,26 in serous carcinomas. Her2/neu was over-expressed in 9,9% of type I and 41,1% of type II ( P< 0,001) and in stage 2 in comparison to stage 1 (54,5% vs 25%). EGFR’s expression was demonstrated in 40,9% and 31,6% of type I and II carcinomas, respectively. Stage 1 type I carcinomas expressed EGFR in statistically significant percentage, compared to stage 2 (77,8% vs 15,4% , P<0,05). CONCLUSIONS: Her2/neu usually is over-expressed in serous type of endometrial carcinoma. Moreover, the expression increases with stage. EGFR especially is expressed in endometrioid type carcinoma and mainly in low stage. Newly developed drugs that target these receptors may have a role in the treatment of endometrial cancer. These results, obtained by immunohistochemistry, have to be confirmed using molecular methods (CISH or FISH).

PP4-22 EXPRESSION OF MASPIN IN TYPE I AND II ENDOMETRIAL CARCINOMA, ITS CORRELATION WITH VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) AND CLINICAL PARAMETERS Maria Sotiropoulou, Irini Papaspyrou, Christos Evaggelakos, Demetra Theocharidou, Pantelis Markoulis, Vassilios Pavlou, Sofia Markaki Histopathology Department , Greece

BACKGROUND: Maspin (mammary–specific serpin) is a tumor suppressor gene that inhibits the invasion and angiogenesis. Maspin is expressed in normal human mammary and prostate epithelial cells but is down –regulated during cancer progression. Interestingly, the protein’s prognostic significance in various cancers is highly impact by its cellular localization (nuclear vs cytoplasmic).Vascular endothelial growth factor (VEGF) plays important role in angiogenesis. The aim of this study is to determine the maspin’s pattern expression in endometrioid (EC) and serous uterine carcinomas (SC), its correlation with VEGF and the clinicopathologic variables. MATERIAL AND METHODS: We examined maspin and VEGF expression in 22 endometrioid and 19 serous uterine carcinomas. Paraffin blocks

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from each case were immunostained, using antibodies against maspin (clone EAW24, NeoMarkers, USA) and VEGF (clone CI 153-694, DD Pharmigen ,USA ) by established methodology. The immunoreactivity of maspin and VEGF was semi-quantitatively scored, based on the intensity and percentage of positive cells (0- +3). oreover, was estimated the maspin’s cellular distribution as cytoplasmic and/or nuclear.. RESULTS: The mean age was 59,8 years for patients with endometrioid carcinoma and 68,3 years for patients with serous carcinoma. Patients' distribution by FIGO grading of EC was as follows: 15 G2 and 7 G3. FIGO surgical stage was as follows: 17 stage I, 6 stage II and 18 stage III. Maspin’s expression was detected in 5/22 (22,7 %)of ECs-5/7 ( 71,4 %) of G3 tumors- and 11/19 ( 58 %, P<0,001)of SCs. In squamous metaplasia of ECs, the stain was absolutely nuclear. There was not correlation between maspin’s expression and vascular invasion on one side and stage on the other. All tumors expressed VEGF (ECs showed lower intensity and had heterogeneity- 36,36 % 1+, 50% 2+, 13,64 % 3+ - despite of SCs which all had strong intensity- 3+). CONCLUSIONS: Maspin and VEGF expression was found in uterine carcinomas related to type ( I or II) and grade. Maspin cytoplasmic expression in contrast to other carcinomas is directly associated with the biological aggressiveness (serous phenotype, and grade) and may result in deregulation of tumor inhibitor properties.

PP4-23 PRIMARY VULVAR EWING’S SARCOMA/PERIPHERAL NEUROECTODERMAL TUMOR: DIAGNOSTIC CONFIRMATION WITH CD 99 IMMUNSTAINING AND REVERSE TRANSCRIPTASE –POLYMERASE CHAIN REACTION Handan Cetiner1, Gozde Kir2 1 Zeynep Kamil Hospital, Pathology Department, Turkey 2 Umraniye Hospital, Pathology Department, Turkey

BACKGROUND: Ewing’s sarcoma (ES) is a rather common, highly malignant primary malignant tumor of bone and soft tissues, and a single pathologic entity with primitive neuroectodermal tumor (PNET). They most commonly involve the deep soft tissues. Situation of this tumor in superficial locations is rare, female genital tract is also an unusual location. We describe a case of ES/PNET occuring as a primary neoplasm in the vulva. To our knowledge, this is the seventh such case reported in the literature METHODS Tumor tissue sections were stained with hematoxylin and eosin, periodic acid schiff (PAS) and PAS-diastase. Immunohistochemical staining for the MIC2 gene product CD99, pankeratin, chromogranin A, synaptophysin, desmin, leucocyte common antigen (LCA), HMB-45 were performed. The t(11;22)(q24;q12) translocation in tumoral tissue was revealed by reverse transcriptase polymerase chain reaction (RT-PCR). CASE REPORT. A 23-year-old woman with 4x4cm left labia majora mass and bilateral pelvic lymphadenopathies is presented. An incisional biopsy revealed a small round cell tumor. The patient underwent left radical hemivulvectomy, inguinal, femoral and paraaortic lymphadenectomy. Tumor was solid, hemorrhagic mass, 6 cm in maximal dimension with 6x4 cm skin ellipse on it. Cut surface was grayish-yellow and friable with infiltrative margins. Diffusely growing cell masses, lobules were seperated by fibrovascular connective tissue.The tumor cells were characterized by dark or vesicular, round nuclei with one to two indistinct nucleoli and scant pale cytoplasm. Immunohistochemical stains revealed that the tumor cells were diffusely immunreactive for vimentin and CD99 (the product of MIC2 gene). Cytokeratin, desmin, HMB-45, LCA, chromogranin, synaptophysin were all negative DISCUSSION: Most frequently extraskeletal ES/PNETs are localized in soft tissues of lower extremities, paravertebral region, chest wall and retroperitoneal region. Gynaecological system is an unusual cite for ES/PNET as is rare involvement of vulva. ES/PNETs have to

be kept in mind in spite of its rarity when small round cell tumor is seen at superficial locations as vulva especially in children and young adults.

PP4-24 IMMUNOHISTOCHEMICAL STUDY OF PAX-2 IN ENDOMETRIAL HYPERPLASIA AND CARCINOMA Maria Sotiropoulou, Irini Papaspyrou, Pantelis Markoulis, Demetra Theocharidou, Christos Evaggelakos, Vassilios Pavlou, Sofia Markaki Histopathology Department, Greece

BACKGROUND: Pax-2 is a paired box gene which has been reported to be activated by estrogen in endometrial carcinoma but not in normal endometrium. However, there are limited studies about Pax-2 expression in endometrial hyperplasia and carcinoma. This study examines the Pax-2 expression in endometrium, aiming to discover any help in the differential diagnosis between atypical hyperplasia and well differentiated endometrioid adenocarcinoma, knowing the major problem in inter observer variability. Moreover, the distinct pattern of Pax-2 expression would be help in the comprehension of endometrial carcinogenesis, related to unopposed estrogen stimulation. METHODS: Immunohistochemical analysis of Pax-2 expression was performed on 57 endometrial specimens: 18 non atypical hyperplasia, simple or complex (SH/CH), 13 atypical hyperplasia (CAH) and 26 endometrioid carcinomas (EC). We used a rabbit polyclonal antibody, Pax-2 (1:50 dilution, Zymed, CA, USA) in established methodology. We estimated only nuclear staining, using a semi-quantitative method as follows: 0=< 5%, 1+=5-25%, 2+=26-50%, 3+=51-100%.Normal endometrial glands, as were uniformly strong positive, were served as positive control. RESULTS: Pax-2 was uniformly expressed in normal endometrium (atrophic or functional). Non-atypical hyperplasia’s epithelial nuclei were positive in 16 cases (88,9%, 2+or 3+) while only in 4 cases of CAH ( 30,76%).Mainly, mild to moderate Pax-2 expression demonstrated in 8 endometrioid carcinomas (30,8%, 4:1+ , 3:2+, 1:3+). CONCLUSIONS: Normal endometriun expresses Pax-2 uniformly and strongly .There is a loss of expression from non- atypical to atypical endometrial hyperplasia and endometrioid carcinoma. The same expression between atypical hyperplasia and endometrioid carcinoma doesn’t permit the use of antibody as a tool in the differential diagnosis .There is a need of further studies in order to be comprehended the relation between Pax-2 expression and ER, the methylation status of the gene and the possibility to convert this polyclonal antibody to a monoclonal one.

PP4-25 PRIMARY INVASIVE SQUAMOUS CELL CARCINOMA OF THE VAGINA: HPV DNA DETECTION, AND P16INK4A, p53, AND PRB IMMUNOEXPRESSION IN 22 CASES Marco Ferreira, Mateus Crespo, Luis Martins, Ana Félix Servico de Anatomia Patológica, Instituto Português de Oncologia de Lisboa Francisco Gentil, EPE, Lisboa, Portugal

Background: Primary invasive squamous cell carcinoma (SCC) of the vagina is rare, and the role of human papilloma virus (HPV) in its pathogenesis remains unclear. Alterations of the cell cycle regulatory proteins p16INK4a, p53 and pRb are poorly known in primary invasive SCC of the vagina. The aim of our study was to investigate HPV DNA, and p16INK4a, p53 and pRb immunoexpression in 22 cases of primary invasive SCC of the vagina. Material and Methods: Tumors (n=22) were classified according to the WHO classification. HPV genotyping (INNO-LiPA HPV Genotyping v.2) was performed in 21 cases. Immunohistochemical staining for p16INK4a, p53 and pRb was assessed in the whole series. Statistical analysis was performed with Fisher’s Exact Test and with Student’s t-test. Results: The

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patients’ age ranged from 36 to 88 (mean 65.17) years. Six cases were keratinizing SCC, and the remaining 16 cases were non-keratinizing SCC (8 “common” non-keratinizing, 3 basaloid, and 5 warty types). The median age of patients with keratinizing SCC was 73.83 yrs. and that of non-keratinizing SCC patients was 62.93 yrs. (p=0.057). HPV DNA was detected in 17 cases (80.9%): 13 (86.7%) non-keratinizing SCC (in one case HPV genotyping was not performed) and 4 (66.7%) keratinizing SCC (p=0.31). The HPV genotypes identified were: 6, 11, 16, 18, 31, 33, 35, 40, and 58. HPV 16 DNA was the most prevalent, being identified in 7 specimens (33.3%); 16 cases (76.2%) had high-risk HPV DNA. In 4 cases (19.0%) multiple HPV genotypes were isolated. p16INK4a was express in 15 cases (68.2%), p53 in 18 cases (81.8%), and pRb in 12 cases (54.5%). Most HPV DNA positive cases were p16INK4a positive (HPV positive: 70.6% vs. HPV negative: 50%), without statistical significance. Likewise, there was no statistical association between HPV DNA detection, and p53 or pRb expression or patients’ age. Conclusions: 1) Similarly to uterine cervix SCC, primary SCC of the vagina is frequently associated with HPV DNA, most cases having high risk HPV DNA, with HPV 16 predominance. 2) There was no statistical difference between HPV DNA detection in keratinizing and non-keratinizing SCC, in contrast to vulvar SCC. 3) p16INK4a expression was not associated with HPV DNA detection. 4) p53 is frequently overexpressed in primary SCC of the vagina.

PP4-26 DIFFUSE OVARIAN STROMAL TYPE METAPLASIA IN TUBAL SUBEPITHELIAL TISSUE Handan Cetiner Zeynep Kamil Hospital, Pathology Department, Turkey

BACKGROUND: Even there were few numbers of reports about ‘ovarian thecal metaplasia in adrenal glands’ in the pathology literature, no case reports describing diffuse ovarian stromal metaplasia of tuba uterina exist. CASE REPORT: A 48-years-old woman, gravida 2, para 2, presented with a 3 weeks history of vaginal bleeding. Pelvic ultrasound (USG) examination revealed a simple adnexal cyst. Also USG revealed 6 cm diameter leiomyomatous mass in uterus. The patient underwent total abdominal hysterectomy bilateral salpingooophorectomy. Grossly: the right fallopian tube was cystic measuring 6cm in diameter and was containing serous fluid.The outer surface of the cystic tube was smooth while inner surface was thick and folded diffusely and had a tan-white color. Other tube and bilateral ovaries had not any specific feature. Microscopically; the stroma of the cystically dilated tuba showed diffusely transformation to dense ovarian stroma under the typical tubal epithelium..There were luteinized stromal cells singly or in the form of small clusters.. These cells were immunreactive for alpha-inhibin. DISCUSSION Other than endometriosis the tubal epithelium may undergo metaplastic changes. Despite of these few cases of epithelial type metaplasia, we couldn’t find any ovarian stromal type metaplasia mentioning report in the literature other than focal fimbrial stromal transformation to ovarian stromal type especially in postmenopausal women.In current case; the change is diffuse and related with hydrosalpinx. To our knowledge, we report the first case of diffuse ovarian stromal type of metaplasia in fallopian tube and suggest that it can be the origin of any type of sex-cord stromal tumors of ovarian type.

PP4-27 MICROVESSEL DENSITY AND IMMUNOHISTOCHEMICAL EXPRESSION OF THE ANGIOGENIC PROTEINS VEGF, VEGFR1 AND VEGFR2 IN NORMAL, HYPERPLASTIC AND NEOPLASTIC ENDOMETRIUM Nektaria Zagorianakou1, Ann Goussia1, Panayiota Zagorianakou1, Emilios Pakos2, Perikles Tsekeris2, Ioannis Nesseris1, Vasiliki Malamou-Mitsi1, Niki J. Agnantis1, Maria Bai1 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Radiation Therapy, Medical School, University of Ioannina, Greece

Background: Vascular endothelial growth factor (VEGF) may play an important role in the multistep process of endometrial carcinogenesis by its effect on angiogenesis and tumor cell proliferation. Therefore, the immunohistochemical expression of VEGF, its specific receptors VEGFR1 and VEGFR2, and the microvessel density (MVD) were analysed in normal, hyperplastic and neoplastic endometrium in relation to MIB1/Ki67 and p53 protein expression as well as to clinicopathological parameters. Materials and Methods: The expression of VEGF, VEGFR1 and VEGFR2 proteins was studied by immunohistochemistry in 31 cases of normal endometrium (15 proliferative and 16 secretory), 36 cases of endometrial hyperplasia (19 simple and 16 complex) and 80 cases of endometrial carcinomas (56 endometrioid and 24 non-endometrioid). MVD was also estimated in all specimens after immunostaining for CD34 antigen and evaluated as the mean (+SD) vessel count/high power microscopic field (HPF). Spearman’s correlation coefficient test was used for the statistical analysis. Results. The expression of VEGF protein was estimated in epithelial cells. In normal endometrium, a significant higher VEGF protein expression was observed in proliferative than in secretory endometrium (p<0.0001). Moreover, VEGF protein expression was significantly higher in hyperplastic than in normal endometrium and in carcinomatous than in hyperplastic endometrium (p=0.002 and p<0.0001, respectively). In endometrial carcinomas, significant positive correlations were found between VEGF/VEGFR1 (p<0.0001), VEGF/VEGFR2 (p=0.049), VEGF/MVD (p<0.001), VEGF/histological grade (p=0.007), VEGF/MIB1 (p=0.005) and VEGF/p53 (p=0.01). Conclusions: The results of this study suggest that VEGF may play an important role in endometrial carcinogenesis by acting a) as a paracrine growth factor promoting angiogenesis and b) as an autocrine growth factor promoting tumor cell proliferation. Moreover, VEGF protein expression may be regulated by p53 in endometrial cancer.

PP4-28 HEPATOID CARCINOMA OF THE OVARY Hulya Ozturk Nazlioglu1, Sema Ozuysal1, Hakan Ozan2 1 Uludag University Medical School, Department of Pathology, Bursa, Turkey 2 Uludag University Medical School, Department of Gynecology and Obstetrics, Bursa, Turkey

Hepatoid carcinoma (HC) is a rare malignant tumour defined as a primary extra-hepatic tumour morphologically mimicking hepatocellular carcinoma (HCC). It has been described in several organs including the ovary.Differentiating this tumor is not only a challenge but also critical, since treatment modalities and operative strategies are dependent upon the exact nature of the hepatoid cancer. We present a rare case of HC of the ovary .A 51-year-old woman was hospitalized with abdominal pain and distention. Physical examination revealed pelvic mass, and abdominopelvic computed tomography showed bilateral ovarian and multiple colonic masses. There was no tumoral mass in the

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hepatic parenchyma, other than capsular penetrations of the colonic tumor. Serum CA 125 level was high (638 U/ml), but serum Alpha Fetoprotein (AFP) was not elevated (1.5 ng/ml). The patient was treated with hysterectomy and bilateral salphingooopherectomy, followed by chemotherapy. Colonic tumours were accepted as inoperable. Histopathologic examination revealed a tumour consisting of large neoplastic cells having vesicular nuclei and abundant eosinophilic cytoplasm resembling hepatocellular carcinoma. Neoplastic cells expressed both CA 125 and AFP .The patient had a poor survival and she died after 20 months following the diagnosis.Based on a review of the literature, we discuss the guidelines for differentiating these tumours and utilize these criteria to differentiate these tumours irrespective of their primary tissue of origin. In conclusion, HC of the ovary should be included in the differential diagnosis of ovarian tumors composed of large eosinophilic cells.

PP4-29 MASPIN EXPRESSION IN ENDOMETRIAL HYPERPLASIA AND CARCINOMA, AND ITS RELATION WITH ANGIOGENESIS Ozlem Erdem1, Cagatay Taskiran2, Cigdem Vural1, Omur Ataoglu1 1 Gazi University Department of Pathology, Turkey 2 Gazi University Department of Obstetrics and Gynecology, Turkey

Background: The purpose of this study was to evaluate the prognostic significance of maspin expression in endometrial hiperplasia and cancer, and also to investigate its relation with angiogenesis. Methods: A total of 19 women with complex atypical hyperplasia, 44 patients with simple hyperplasia without atypia, and 67 patients with endometrial carcinoma were included. Maspin expression was assessed by immunohistochemistry and tested for possible significant relation with age, FIGO stage, histologic type, grade, depth of myometrial invasion, lymphovascular space involvement, lymph node metastasis, and overall survival . Angiogenesis was determined by vascular endothelial growth factor staining. Results: Maspin expression was detected in only three patients with endometrial hyperplasia. In patients with endometrial cancer, cytoplasmic and nuclear maspin expressions were detected in 36 (53.7%) and 18 (26.9%) patients, respectively. No significant relation was noted between prognostic variables and staining localizations. The 5-year OS rate for patients with cytoplasmic staining was 91%, compared to 87% for patients without staining (p=0.31). The values for nuclear expression were 100% and 87%, respectively (p=0.16). The cytoplasmic and nuclear maspin expressions were found to be significantly correlated with VEGF (r=0.278, p=0.02; and r=0.295, p=0.02, respectively). Conclusion: This is the first study investigating the relation between maspin expression and angiogenesis in endometrial cancer. Although no significant correlation was noted between the expression of maspin and clinicopathologic prognosticators and no survival difference was noted for cytoplasmic and nuclear expression of maspin, a tendency was detected for nuclear staining similar to the literature. Further series will clarify the exact prognostic role of maspin in gynecologic malignancies including endometrial cancer.

PP4-30 RETROSPECTIVE ANALYSIS OF GYNECOLOGICAL SARCOMAS: Alp Usubutun1, Cigdem Himmetoglu1, Guldeniz Aksan2, Sertac Esin2, Ali Ayhan2, Turkan Kucukali1 1 Hacettepe University Faculty of Medicine Department of Pathology, Turkey 2 Hacettepe University Faculty of Medicine Department of Gynecology and Obstetrics, Turkey

Background: Gynecological sarcomas, comprised mostly of uterine origin, are rare and generally care a poor prognosis. Histological prognostic features are confusing due to absence of large patient data. The aim of this study was retrospective re-analysis of all the gynecological sarcomas in our institution to evaluate the morphological prognostic factors. Method: Out of 182 cases treated in our institution with gynecological sarcoma diagnosis between 1968-2007, 149 cases were available for pathological examination. Clinical data were obtained from patients’ medical records. All cases were examined by the same pathologist. The histologic type, presence and percentage of necrosis, mitotic count per 10 HPF, the grade of atypia, cellularity, lymphovascular space involvement (LVI) and lymph node metastasis (LNM) were evaluated. Survival analysis (Kaplan-Meier) and long rank test were used for statistical analysis. A new scoring system was developed by combining mitotic count (1-5=1, 6-10=2, >10=3), percentage of necrosis (<25%=1, 25-50%=2, >50%=3) and nuclear atypia (mild=1, moderate=2, severe=3). Results: Of the 149 gynecological sarcoma cases, 49.7%(74) were leiomyosarcoma (LMS), 32.2%(48) were carcinosarcoma and 10.1%(15)were endometrial stromal sarcoma (ESS). The uterus was the site of origin in 86.6%(129) of cases. The age at diagnosis varied between 16 - 84 years (mean: 53). Patients were followed-up for a period of 1 to 197 months (mean: 26.5). Considering all the sarcoma cases clinical stage, histologic type, grade of atypia, percentage of necrosis, LNM and mitotic rate were found to be statistically significant for survival (p<0.005). Necrosis and LNM were found to be the prognostic parameters among LMS cases; whereas in carcinosarcomas necrosis, LVI and LNM emerged as statistically significant factors. In addition, the analysis of LMS cases using the newly developed scoring system was found to be a valuable prognostic factor (p=0.0004). Conclusion: Whatever the histologic type and site of origin, this study shows that mitotic rate, percentage of necrosis and grade of atypia are the histological prognostic factors-with varying influence-in gynecological sarcomas. Besides, LVI and necrosis for carcinosarcoma, and necrosis for LMS come out to be the the prominent and statistically significant factors. There appears that no single histological factor is enough to determine the prognosis in gynecological sarcomas. However, this scoring system, combining some of the morphological features, was found to be a valuable prognostic factor for LMS.

PP4-31 HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL CORRELATIONS BETWEEN ADENOMYOMATOUS POLYPS, HYPERPLASTIC POLYPS AND ENDOMETRIAL MALIGNANCIES Cornelia Amalinei, Raluca Balan, Irina Draga Caruntu, Corina Cianga, Petru Cianga, Stefan Butureanu, Alexandra Pangal University of Medicine and Pharmacy, Romania Background: Endometrial polyps are considered markers of increased cancer risk, revealing endometrium tendency to develop proliferative lesions. Beside well recognized precursors of endometrioid endometrial carcinoma represented by diffuse endometrial hyperplasia and, in a lesser degree, by hyperplastic endometrial polyps, when associated with cytological atypia, a large interest in recent years has centered on endometrial polyps with myomatous stroma, including those with atypical features. Polypoid adenomyomas cover a large spectrum of proliferative benign and premalignant changes, being designated as low malignant potential lesions. The aim of our study was to investigate the correlations between histopathological and immunohistochemical features of these neoplasias. Method: As a material, we used 46 polypoid adenomyomas and hyperplastic polyps, selected from our files after excluding frank cancers, conventional polyps, functional polyps, atrofic polyps, mixed

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endometrial-endocervical polyps, and decidual pseudopolyps. Results: Histopathological examination identified cytological atypia and associated areas of malignancy. Immunohistochemical staining for ER, PR, MMP-2, MMP-9, Ki-67, and cyclin D1 revealed a significant correlation with hormonal status, expression of stromal components and proliferative activity in cases that associated great cytologic atypia and malignant transformation respectively. Differential diagnosis was difficult due to the typical fragmentation of curretage specimens and included florid hyperplasia, adenomyoma, adenocarcinoma with polypid growth, and mesenchymal tumors. Conclusion: Correct identification of stromal and cellular alterations often provides a crucial clue to the therapeutic orientation and assures prognostic improvement. Identification of histopathological and immunohistochemical markers of intense proliferation associated with invasion activity is important in positive and differential diagnosis with carcinomas, carcinosarcomas and adenosarcomas. PP4-32 EXPRESSION OF EGFR AND C-KIT (CD117) IN UTERINE LEIOMYOSARCOMAS Delia Perez Montiel1, David Cantu De Leon1, Jose Luis Aguila Ponce1, Saul Suster2, Jose Chanona Vilchis1 1 Instituto Nacional de Cancerologia, Mexico 2 Ohio State University Medical Center, Mexico Objectives: Determine the expression of epidermal growth factor receptor (EGFR) and a growth factor receptor with tyrosine kinase activity, c-kit (CD117), in uterine leiomyosarcoma and correlate with clinical and pathologic characteristics. Materials and Methods: Twenty-seven cases of uterine leiomyosarcomas were immunohistochemically evaluated for the expression of EGFR and c-kit (CD117) and correlated with clinical and pathological features. Results: EGFR was positive in 13 cases (48.1%), 71% of patients were younger than 50 years (p=0.002). Statistical analysis showed that expression of EGFR was not an independent predictive factor for recurrence (p=0.22). EGFR expression was also not related to clinical stage or other clinical and pathological features (p=0.7). Mean survival in cases positive for EGFR was 17 months in comparison with EGFR negative cases, for which mean survival was 60 months (p=0.068). C-kit (CD117) was positive only in one case of uterine leiomyosarcoma (3%), therefore, no valid statistical conclusions could be established. Conclusions: Treatment of uterine leiomyosarcomas is generally poor; the use of targeted therapies may help to improve long term control and survival. Expression of two specific targets for cancer therapy, EGFR and C-kit, did not appear to demonstrate any specific correlation with survival or clinical and pathological features in uterine leiomyosarcomas. PP4-33 HISTOGENESIS OF LIPOMATOUS COMPONENT IN UTERINE LIPOLEIOMYOMAS Filiz Bolat1, Fazilet Kayaselcuk1, E. Tuba Canpolat1, Serkan Erkanli2, Ilhan Tuncer3 1 Baskent University Faculty of Medicine, Department of Pathology, Turkey 2 Baskent University Faculty of Medicine, Department of Gynecology and Obstetric, Turkey 3 Baskent University Faculty of MedicineDepartment of Pathology, Turkey Background: Uterine neoplasm composed of an admixture of smooth muscle and adipose tissue are relatively common and have been designated as lipoleiomyomas. The origin of this tumor is still controversial and it has not been sufficiently studied. Aim of our research was to investigate the immunohistochemical phenotype of the fat cells in uterine lipoleiomyomas for clarifying their origin. Methods: Archived tissue samples of 10 uterine lipoleiomyomas were selected and analyzed immunohistochemically for vimentin, desmin, and

HMB45 expression. Results: The patients ranged from 31 to 63 years of age (mean age 53.5±9.9). Seven tumors affected the uterine corpus, and were located intramural; two cases were subserosal, and the one was in the cervix. All tumors were constituted by irregular bundles of smooth cells and mature large adipose cells. The amount of adipose component varied from 5 to 95 % of the tumor mass. Cytological atypia and necrosis were not seen. The immunohistochemical investigations revealed obvius reactivity to vimentin and desmin in perivascular and tumor smooth muscle cells. Adipose cells in the tumors demonstrated uniformly vimentin expression and inconsistent desmin immunreactivity. All adipose cells were negative for HMB45 antigen. However, HMB45 antigen was weakly positive in spindle shaped tumor cells of two cases. Conclusions: The immunohistochemical findings suggest a complex histogenesis for these tumors, which may arise from perivascular immature mesencyhmal cells or direct transformation of smooth muscle cells into adipocyte by means of progressive intracellular storage of lipids. PP4-34 VULVAR CARCINOSARCOMA: A CASE PRESENTATION Jasmina Atanackovic, Svetlana Milenkovic, Jasmina Tadic, Milena Cosic Micev, Marjan Micev Clinical Centre of Serbia, Department of Histopathology, Belgrade, Serbia Vulvar carcinosarcoma is extremely rare and the least frequent of all genital localizations with just a few reported cases. In the limited data in the literature, the additional problem of estimating the rarety of this tumour represents inconsistent terminology of these neoplasms. Carcinosarcoma in the genital tract, often reported as malignant Mullerian mixed tumour, in most cases was accepted as being metaplastic or sarcomatoid carcinoma. A proportion of these cases in the vulvar region were confirmed as keratin-positive sarcomatoid carcinoma or squamous cell carcinoma with spindle cell sarcoma-like transformation of the stroma. However, carcinosarcoma is defined as mixed epithelial and mesenchymal tumour with malignant both epithelial and mesenchymal component, either homologous or heterologous by cellular origin. Recently, the clonal origin of the first case of squamous carcinomatous and leiomyosarcomatous diffefrentiation in the same tumour has been genetically proved. We report a case of vulvar carcinosarcoma diagnosed in a 66 year-old woman who underwent radical vulvectomy with bilateral inguinal lymphadenectomy because of rapidly grown tumour in the right labial region. Histopathological analysis presented ill-defined mass measuring 70 mm in maximal diameter and revealed the admixture of well differentiated squamous cell carcinoma and distinctive spindle cell sarcomatous areas with fibrosarcoma-like aspect. Final report showed FIGO II stage without deposits in any of 14 lymph nodes. Immunohistochemical analysis were performed on 4 microns sections from formalin fixed and paraffin-embedded samples using LSAB+ immunostaining method with AEC visualization for: vimentin, smooth muscle actin - alpha, desmin, S-100 protein, CD34, HHF35, GFAP, CD117, NSE, EMA, AE1/AE3, High molecular weight cytokeratins, p53 protein, and proliferative activity measured by Ki-67 protein labelin index. Morphologically readily seen sarcomatous component immuno-histochemically showed the presence of strong cytoplasmic positivity only for vimentin, contrasting to strong positivity for all epithelial markers in the carcinoma; Ki-67 protein labelling index was 42% and immunopositivity for p53 (about 55%). In conclusion, we believe that morphological and immunohistochemical characterizations of two neoplastic populations represent true carcinosarcoma. Besides differential diagnostic significance, it remains unclear whether biclonality or sarcomatoid metaplastic carcinomatous or stromal transformation bear more unfavourable prognosis in these aggressive tumours.

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PP4-35 P53, COX-2 EXPRESSIONS AND MICROVESSEL DENSITY IN OVARIAN CARCINOMAS Aysel Caglar1, Elife Sahan2, Aysenur Akyildiz Igdem2, Nusret Erdogan2 1 Bagcilar Education and Training Hospital, Pathology Department, Istanbul, Turkey 2 Taksim Education and Training Hospital, Pathology Department, Istanbul, Turkey Ovarian carcinomas are the sixth of the most common carcinomas. Surface epithelial and stromal tumors are the most common neoplasms in ovarian carcinomas. In molecular studies, the mutation of p53, which is a tumor supressor gene, is positive in %50-60 of invasive serous carcinomas. Neoangiogenesis is an important factor in carcinogenesis. The growth of tumor is thought to be prevented by antagonists of angiogenesis and it can be an effective treatment method. Cyclooxygenase is an important catalyser enzyme in the synthesis of prostoglandins from arachidonic acids. It is suggested that by the epidemiological studies, inhibition of cox-2 with the help of nonsteroidal antiinflamatuar drugs can prevent ovarian carcinomas. We studied on 17 primary serous adenocarcinomas, 3 primary musinous cystadenocarcinomas, 6 borderline serous tumors, 4 borderline musinous tumors,10 serous cystadenomas,10 musinous cystadenocarcinomas retrospectively and researched clinical and histopathological features. For cox-2 expression, the difference between musinous tumors and serous tumors was meaningful.For p53 positivity, the difference between malignant, borderline and benign ovarian tumors and serous versus musinous tumors was meaningful. The difference between malignant, borderline and benign ovarian tumors for MVD>60 was meaningful. There was no correlation between cox-2, p53 and MVD and survival. For reducing the diameter of tumors by neoadjuvant and adjuvant therapies, the methods for reducing the vascularity can be useful. For the treatment of ovarian carcinomas, the experimental studies of p53 gene therapy should be developed. In current pathology practice; in ovarian tumors suspicious for malignancy by the histomophological findings; the positivity of p53, high microvessel density, old age, bilaterality are the helpful factors for differentiating malignant and borderline tumors. For evaluating the role of cox-2 in ovarian tumors there is need for larger series of cases which are chemotherapy resistant and /or chemotherapy responsive in oncology services. PP4-36 ATYPICAL IMMATURE METAPLASIA OF CERVIX- THE SIGNIFICANCE OF KI-67 IN DIFFERENTIAL DIAGNOSIS WITH SQUAMOUS INTRAEPITHELIAL LESIONS Ovgu Aydin, Ahu Senem Demiroz, Haydar Durak, Nuray Kepil, Sennur Ilvan, Zerrin Calay Department of Pathology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey Background: Atypical immature metaplasia (AIM) of cervix is an epithelial alteration with uncertain biological and clinical significance. The main issue is to differentiate AIM from squamous intraepithelial lesions, especially high grade squamous intraepithelial lesion (HGSIL) with which it has many morphological similarities. The histological criteria of AIM are a uniform basal cell population, minimal nuclear crowding, variable hyperchromatism, preserved cellular polarity, enlarged or multiple nuclei confined to suprabasal areas and absence of abnormal mitoses. Method and Result: In our study histological feaures and Ki-67 proliferation index were evaluated in the cervical biopsy specimens from 23 cases; including 5 AIM , 6 low grade squamous intraepithelial lesions (LGSIL), 12 HGSIL. The mitotic index and Ki-67 counting in the basal one-third, middle one-third and superficial one-third of epithelium were

assessed seperately.For Ki-67 immunoreactivity percentage, at least 200 cells from informative areas were evaluated. The mean Ki-67 indices of AIM, LGSIL and HGSIL were 10,4; 7,9; 37,3 respectively. Overall Ki-67 index was higher than that of LGSIL and lower than HGSIL though the range of Ki-67 indices of AIM overlapped that of both LGSIL and HGSIL. When the Ki-67 indices were evaluated for each 1/3 layer seperately; the discriminative values were found to be for upper and mid third of epithelium rather than the basal third. When the histological features and the Ki-67 indices were evaluated together; 1 biopsy which was thought to be AIM turned out to be HGSIL. Conclusion: AIM is a poorly defined entity and the pathologist can have some hard time when dealing with this lesion especially in the differential diagnosis with HGSIL. Histological criteria are inadequate in such situations and Ki-67 index should be used in assesment of such atypical proliferations of cervix . PP4-37 COMPARISON OF ESTROGEN, PROGESTERONE EXPRESSION AND ANGIOGENESIS WITH OTHER PROGNOSTIC PARAMETERS IN ENDOMETRIAL CARCINOMAS Serap Koc, Nimet Karadayi, Dilek Yavuzer, Nagehan Ozdemir Barisik Dr. Lutfi Kırdar Kartal Educational and Research Hospital, Department of Pathology, Turkey BACKGROUND: Besides the established prognostic parameters in endometrial carcinomas, recent researches revealed that estrogen, progesterone status, angiogenesis, HER-2/neu expression and tumour DNA ploidy have significant prognostic value. This study investigates the relationship between estrogen, progesterone receptor expressions, microvessel proliferation (angiogenesis) and established prognostic parameters. METHOD: Sixty two endometrial carcinoma diagnosed at Dr. Lutfi Kırdar Kartal Educational and Research Hospital are included in this study. Estrogen, progesterone and CD34 receptors are assessed immunohistochemically at sections from the blocks which are the best representative of histological grade, which have the deepest myometrial invasion and in which stroma can be viewed easily. Results are compared with other prognostic parameters including patient age, tumour type, histological grade, myometrial invasion, vascular invasion, lymph node metastasis, endocervical involvement and stage. RESULTS: A statistically significant difference is found between estrogen receptor positivity and tumour type or histological grade (p<0.01). A statistically significant difference is found also between progesterone receptor positivity and tumour type or grade (p<0.01). Any significant relation is not established between either receptors and other parameters. A significant relation is not found between microvessel proliferation and other prognostic parameters. Although a consistency is seen between estrogen and progesterone expressions, they do not have a significant relation with angiogenesis. CONCLUSION: For the endometrial tumours whose clinical behaviour can not be precisely assessed pre-operatively, the contributions of prognostic parameters investigated post-operatively are important. However, all of the cases do not act in accordance with available prognostic parametres. Such cases act more agressively. The aim is to detect those cases during planning treatment and step up to a more agressive treatment plan. There are several parameters in literature and we chose to assess steroid receptor status and angiogenesis. Although we found significant relation between hormon receptor status and tumour type and histological grade, unlike some reports, we could not find any relation with angiogenesis

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PP4-38 A MOLECULAR STUDY ON SELECTED BRCA-1 AND BRCA-2 GERMLINE MUTATIONS IN BREAST, OVARY AND FALLOPIAN TUBE CANCER CASES IN TURKEY E. Handan Zeren1, Elif Guveloglu1, Melek Ergin1, Demet Aras1, Derya Gumurdulu1, Fatma Bayram1, M. Ali Vardar2, Orhan Demircan3. 1 Cukurova University, Faculty of Medicine, Dept. of Pathology, Adana, Turkey 2 Cukurova University, Faculty of Medicine, Dept. of Gynecology, Adana, Turkey 3 Cukurova University, Faculty of Medicine, Dept. of Surgery, Adana, Turkey

Background: Germline mutations in BRCA1 and BRCA2 genes increase the susceptibility to ovarian, peritoneal, fallopian tube, and breast cancers. 185delAG and 5382insC in BRCA1 and 6174delT in BRCA2 are well known, common mutations among Ashkenazi Jewish people and as frequent as 2.6% of the population. In a background of the mosaic Turkish population and its close relationship with Middle Eastern countries, we aimed to detect these three mutations in a series of young cancer patients. Materials and Methods:A total of 112 histologically confirmed breast (invasive ductal carcinomas), ovary, and fallopian tube (serous carcinomas) cancer patients under the age of 45 regardsless their family history are included. All cases were genetically tested for the three BRCA1-2 founder germline mutations (185delAG and 5382insC in BRCA1 and 6174delT in BRCA2). The analysis was performed on genomic DNA extracted from paraffin embedded tumor tissues. We used a simple and rapid method for the simultaneous detection of these three mutations. Allele-specific oligonucleotide primers designed on the basis of BRCA1 and BRCA2 sequences were used. Mutation in one of the alleles was detected by the presence of two bands. For the 185delAG mutation, the mutant and wild-type amplicons were 354 and 335 bp, whereas those of 5382insC were 295 and 271 bp, and those of 6174delT were 171 and 151 bp. Results:All breast carcinoma cases were negative for the three mutations. Among others, a case of ovarian serous carcinoma from a 42-year old woman, showed mutation in BRCA1 5382insC. The patient had no history of familial ovarian and/or breast cancer. The tumor was unilateral and revealed high grade morphologıc features. Conclusion: Detection of a BRCA1 mutation in a series of younger ovarian cancer patients is an important finding and may point more mutations among the population. PP4-39 ADULT GRANULOSA CELL TUMOR OF THE OVARY: STUDY OF 20 CASES Maurício De Angelo Andrade, Joyce De Brito Pupo, Liliana De Angelo Andrade Universidade Estadual de Campinas, Brazil

Background: Among various ovarian neoplasms, granulosa cell tumors of the adult type (GCT) correspond to less than 5%. They have low malignancy and are capable of recurring even after many years. They present many diverse histological aspects, whose differential diagnoses must be made with other primary or metastatic tumors in the ovary. Objective: to describe and to analyze clinical and pathological aspects of GCT and relate them to the evolution of the disease. Method: in a 10-year (1995-2004) review of the UNICAMP Clinical Hospital files, twenty CGT diagnoses were found, and these records were reviewed, correlating clinical aspects with morphological patterns, mitotic index, stage of the disease and frozen section diagnosis accuracy. Results: the ages varied from 27 to 79 years (mean: 53 years) and the follow-up from 12 to 96 months (mean: 42). The main symptoms were: postmenopause bleeding (45%), abdominal pain (35%) and palpable mass (25%). In gross analyses, most of the tumors were yellowish (60%) and the solid aspect was more common (40%) than the cystic or solid-cystic. Only three of the

nine cases submitted to intra-operative frozen section biopsies were diagnosed as GCT (33%). The histological patterns were: 40% solid, 15% macrocystic and 45% of combined patterns, with low mitotic index. The clinical stage was: 13 cases in Ia (65%), 2 cases in Ic and 5 in IIIc. Only the stage of the disease was related to the recurrences. In 3 of 14 hysterectomies there was simple endometrial hyperplasia without atypia. Conclusion: The diagnosis of CGT generally occurs after the menopause, and intra-operative biopsies are seldom conclusive. Only the advanced stage was related to the worst prognosis. PP4-40 DIFFUSE LARGE B-CELL NON-HODGKIN LYMPHOMA (DLNHL) - MIMICKING A PRIMARY GYNECOLOGICAL TUMOR Ljiljana Tomic1, Vlado Jeremic2, Olga Rdaic-Tasic3, Milena Djukic4 1 Department of Pathology, The Clinic for Obstetrics and Gynecology “Narodni Front”, Belgrade, Serbia 2 Department of Surgical Gynecology, The Clinic for Obstetrics and Gynecology “Narodni Front”, Belgrade, Serbia 3 Department of Pathology, Medical Military Academy, Belgrade, Serbia 4 Department of Pathology, Mother and Child Health Institut "dr Vukan Cupic", Belgrade, Serbia

BACKGROUND: Bilateral ovarian involvement mimicking a gynecologic malignancy in adults is extremely rare. Here, we report a patient with DLNHL, mimicking a gynecologic tumor. Aim: To determine the origin of the tumor, using immunophenotyping, with the introductory help of cytological imprints that had been taken simultaneously with the intraoperative frozen-section tissue samples. The very cytological imprints gave the clue way and reveal the possible true origin of the tumor-cells, differentiating lymphoblasts from others like “small round blue cell”- tumors. METHOD: A 57 year old Serbian rural woman, presented with the complaint of persistent lower abdominal distension with dull pain, and night sweats. Imaging studies revealed a huge ovarian mass, suggesting a large ovarian tumors. Surgery-laparatomia with total hysterectomia and bilateral adnexetomia and total omentectomia with intraoperative frozen-section analyses was performed. DLNHL diagnosed by a combination of: cytology, morphology and immunophenotype together with clinical features (“B-symptoms”-associated with lymphoma), managed with surgery and chemotherapy (MABTHERA). Subsequently patient staged, according Ann Arbor classification-IV B. RESULTS: The average diameter of the ovaries was 16 cm (range 12-20 cm), grossly- lobular surface, pink-grey color, soft consistency. Microscopic findings: ovaries, myometrium as well as peritoneal implants were completely replaced by the monotonous, uniform population of tumor cells-centroblasts. By the use of IHH: tumor cells have shown excessive immunopositivity towards CD20, CD79a, Ki67 (80%) and bcl2 (40%) while negative towards CD3, CD5, CD138, MPO, ALK and CK. CONCLUSION: Although rare secondary ovarian tumors i.e.lymphomas should be considered in the differential diagnosis of female adults with ovarian masses. Bilateral ovarian masses or large masses indistinguishable from the ovaries, particularly in the presence of other metastatic foci (secondary deposits in lungs), may help distinguish primary from secondary ovarian malignancies. PP4-42 PREDICTORS OF RESIDUAL DISEASE ON HYSTERECTOMY AFTER LOOP ELECTROSURGICAL EXCISIONAL PROCEDURE WITH POSITIVE MARGINS David Cantu De Leon1, Delia Perez Montiel1, Diana Copado Mendoza2, Olivia Gomez Alvarado2, Jose Chanona Vilchis1 1 Instituto Nacional de Cancerologia, Mexico 2 Instituto Tecnologico y de Estudios Superiores de Monterrey, School of Medicine, Mexico

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Background: Loop electrosurgical excisional procedure (LEEP) is the treatment of choice in patients with cervical intraepithelial neoplasia (CIN). Positive surgical margins may be the most important prognostic factor for residual disease and patients have to undergo hysterectomy for definitive treatment with a rate as high as 50% of specimens negative. This study was performed in intent to identify predictors for residual disease in hysterectomy specimens of patients with positive margins in LEEP specimens. Material and methods: Pathological files were reviewed from 1998 to 2003. Cases with LEEP and positive margins that were undertaken to hysterectomy were identified. Histopathologic slides from the LEEP and hysterectomy were reviewed to confirm margin status, grade of lesion, and residual neoplasia. Variables such as age, parity, number of sexual partners, time from LEEP to hysterectomy, size of lesion, percent of affected fragments, and number of positive margins (endocervical, exocervical, or both) were analyzed. Univariate analysis was performed using Chi-square test, t test, and Wilkoxon Rank Sum. Multivariate logistic regression analysis was used to test relationships between preoperative factors and presence of residual disease on the final pathologic report. Results were considered statistically significant if p < 0.05. Results: 98 specimens were evaluated, median age was 42 years ( range 25-78 ), age at first intercourse 17y ( range 12-34), number of sexual partners 2 ( range 1-10), parity 5 ( range 1-16), and days prior to hysterectomy 48 days. Of LEEP specimens median size of lesion was 0.5 cms (range 0.1 to 2cm.). Median of percent of fragments affected was 40% (5-100%). Exocervical involvement was present in 6%, endocervical 64%, both 30% respectively. Median number of fragments with endocervical margins affected 3 ( range 0-11 ), exocervical 0 ( range 0-10). Of hysterectomy specimens 56 (58%) were negative residual disease, 2 (2.1%) CIN II, 2 (2.1%) CIN III, 31 (32.3%) In situ carcinoma, 2 (2.1%) microinvasive, 3 (3.2%) invasive carcinoma. On multivariate analysis variables that independently predicted residual disease were age ( OR= 1.049, p= 0.05) and percent of affected fragments ( OR= 9.74, p=0.038 ). Conclusion In our institution we have a high rate of negative hysterectomy specimens which is similar to other reports. Age and extent of disease (percent of affected fragments) are statistically significant predictors of residual disease rather than positiveness of margins as well as number of positive margins. PP4-43 RARE TYPES OF CARCINOMAS LOCALIZED IN THE ENDOMETRIUM Raluca Balan1, Cornelia Amalinei1, Irina Draga Caruntu1, Vlad Gheorghita2 1 Morphology Department, University of Medicine and Pharmacy, Iasi, Romania 2 The 3rd Clinical Hospital of Obstetrics and Gynecology, Iasi, Romania Background: There have been reported rare examples of unusual carcinomas arising in the endometrium, but the data consist largely of case reports precluding a detailed clinicopathologic analysis. Material and method: We present 3 cases of endometrial carcinomas, each of them with different clinicopathologic features. The specimens were surgically treated, routinely processed and stained with H&E, PAS, van Gieson, and a panel of immunohistochemical antibodies. Results: One case was histopathologically diagnosed as synchronous carcinoma of the endometrium and uterine cervix, one was a metastatic mucous adenocarcinoma in the endometrium, primary located in the ovary, and the last one was diagnosed as poorly differentiated endometrioid carcinoma of the endometrium, developed in a background of inflammation. Conclusions: Cervicitis, salpingitis, endometritis, and myometritis may represent an important background for the development of an endometrial carcinoma. Immunohistochemical studies may be helpful for distinguish a metastatic carcinoma from an independent tumor, but the

differential diagnosis usually can be determined by conventional clinicopathologic criteria. The distinction is very important because the prognosis and treatment are different. PP4-44 REAPPRAISAL OF OVARIAN METASTASIS FROM GALLBLADDER AND EXTRA-HEPATIC BILE DUCT CARCINOMA Yoji Wani, Kaori Uchino, Kenji Notohara, Choutatsu Tsukayama Kurashiki Central Hospital, Japan Background: Metastatic ovarian tumor derived from gallbadder or extra-hepatic bile duct (GB/BD) is estimated about 7% in overall ovarian metastasis. Especially, tumor-forming metastasis (classical or tubular Krukenberg tumor; KT) is extremely rare and most of all are limited in case reports. The aim is to reappraise ovarian metastasis originating in GB/BD. Methods: From autopsy files, 40 female GB/BD carcinomas were reviewed. We evaluated presence of the ovarian metastasis, number of other metastatic organs, presence of peritoneal dissemination, histology. Furthermore, 11 KTs from eight (7; surgical) cases were clinicopathologicaly evaluated. Results: Patients were 56 to 97 years of age. In 12 (30%) cases, ovarian metastasis was identified (bilateral: 4, unilateral: 8). The mean number of involved organs other than ovary was 4.42 (0-7) in contrast to 2.96 (0-6) in cases without ovarian metastasis. 11 cases were involved by peritoneal dissemination. All but one ovary was macroscopically normal or atrophic. Only one (1/12: 8.3%) KT, which measured 5x4 cm, showed multilocular appearance. Histologically, the metastatic cases consisted of 8 tubular adenocarcinomas, 2 signet-ring cell carcinomas. The rest was one case of clear cell, papillary adenocarcinoma, respectively. KT patients were 49 to 71 years of age. Five primary sources were gallbladder and three were bile ducts. With surgical cases, two were at stage II, III, respectively other than three at stage IV. The duration between biliary surgery and oophorectomy were 5 to 68 months. The laterality was shown as unilateral; 3 as bilateral; 5. In 11 ovarian tumors, the specimens ranged from 3.1 to 17 cm in greatest diameter and four were just described such as newborn’s or adult head’s size. They showed predominantly solid (4), solid and cystic (2) and multilocular (5) appearance. Eight tumors composed of tubular adenocarcinoma (one with foci of signet-ring cells) and each one was papillary adenocarcinoma and signet ring cell carcinoma. All cases were associated with cancerous ascites or peritoneal dissemination. From 7 to 12 months (mean; 9.8) after oophorectomy, 5 patients were died of disease and two was alive with disease for 12, 13 months, respectively. Conclusion: Ovarian metastasis from GB/BD carcinomas is not unusual at the advanced stage, but rare with KT. Classical KT is rare while most cases are tubular ones. KTs may be early or primary manifestation of metastasis in some cases, but frequently associated with peritoneal dissemination, the prognosis is poor. PP4-46 PERITONEAL INCLUSION CYSTS: A CLINICOPATHOLOGIC STUDY OF 20 CASES Metin Akbulut1, Osman Zekioglu2, Mustafa Cosan Terek3, Necmettin Ozdemir2 1 Pamukkale University, Department of Pathology, Denizli, Turkey 2 Ege University, Department of Pathology, Izmir, Turkey 3 Ege University, Department of Obstetrics&Gynecology, Izmir, Turkey Background: Peritoneal inclusion cysts are uncommon mesothelial proliferations that typically occur in the peritoneal cavity of women in the reproductive age. It was reported that they have tendency for recurrence, however; malign transformation has never been reported in the literature. This study was planned to determine the clinicopathologic features of peritoneal inclusion cysts. Methods: All patients with a diagnosis of peritoneal inclusion cyst were studied retrospectively from a retrospective

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database at Ege and Pamukkale University Hospital between January 1995 and March 2007. Results: Twenty patients with a confirmed diagnosis of peritoneal inclusion cyst were identified. All the patients were women ranging in age from 20 to 56 years old (mean 37 years). The associated clinical conditions were endometrioid adenocarcinoma for 1 patient, endometriosis for 3 patients, mature cystic teratoma for 2 patients, pregnancy for 4 patients, leiomyoma of the uteri for 7 patients. The commonest location of the tumor mass was below the pelvic diaphragm. The size of the tumor was larger than 5 cm in 5 patients. The tumors presenting in the peritoneum were multilocular and large. The rest of the tumors were small. The septa consisted of loose fibrovascular tissue. No smooth muscle and no significant amount of inflammatory cells were present. Microscopically, the majority of cysts were lined with a single layer of flattened or cuboid mesothelial cells. These cells stained positively for calretinin and were negative for endothelial marker CD31. Conclusion: Peritoneal inclusion cyst is a rare and distinctive lesion of mesothelial origin. Most of the patients have a history of a previous pelvic operation, and endometriosis without recurrence, suggesting a role for inflammation in the pathogenesis of the cysts as a peritoneal reactive proliferation rather than the neoplastic process. PP4-47 “FETAL-PLACENTAL UNIT”: PROTOCOLIZED STUDY OF 2847 PLACENTAL SPECIMENS AND 432 FETUS IN THE LAST 7 YEARS. Ana Puras1, JM Ezpeleta2, A López Cousillas1, A Echegoyen1, P Fernandez Seara1, Y Ruiz de Azua1 1 Departments of Pathology Hospital Virgen del Camino Pamplona, Spain 2 Departments of Obstetrics Hospital Virgen del Camino Pamplona, Spain Introduction: Both placenta and fetus have a common origin (Fetal- Placental Unit) for that reason it should be studied together in cases of fetal death. In other cases, obstetric and clinical information should be carefully recorded. In this condition, placental examination and their study will be useful for patients, obstetricians and pediatricians. The indications for placental examination should be recomended in diverse causes of fetal, placental, maternal u obstetric disorders possibles of maternal or fetal problems. This could be use to prevent and control pathologies in following pregnancies. Material and Methods: In our Pathology Department have been examined 2847 Placentas since year 2000. They were received fresh and the handling of the placentas was according to a stablished protocol that includes: reception, description, cultures, frozen sections of umbilical cord, postdelivery drainage, membranes roll, disc weight after removing cord, membranes and clots, formalin fixation, disc sectioning and standarized sampling for microscopic examination. Fetus were all autopsied, Rx taken, photographs if needed and any anomaly described. Results: Every pathology report was sent to the Obstetrician and Perinathology Departmens, and those with funiculitis in frozen section by urgent shipment. Every autopsy report included the placental report, and a summary about the correlation of findings. The fetal reports were also sent to the Obstetrician and Perinathology Departments, and rewied in the Clinical- Pathological Conference. Conclusions: 1- The application of this Protocol developed in consensus with the Obstetrics Department, has been very useful in the clinical-pathological correlation of the feto placental pathology, and in the handling of the obstetrical pathology in following pregnancies. 2- Frozen sections of umbilical cord have been useful in early detection of fetal inflammatory response and adverse neonatal outcome.

PP4-48 PREVALENCE AND CHARACTERISTICS OF ENDOMETRIAL POLYPS IN PATIENTS WITH ABNORMAL UTERINE BLEEDING Biljana Djordjevic, Ljubinka Velickovic, Vesna Zivkovic, Miljan Krstic, Miljan Milic Institute of Pathology, Medical Faculty, University of Nis, Serbia Background/Aim. The prevalence of endometrial polyps in the general female population is about 24%. Abnormal uterine bleeding is frequently the presenting symptom of endometrial polyps. The aim of this study was to determine prevalence and characteristics of endometrial polyps in patients with abnormal uterine bleeding. Methods:. The prevalence and characteristics of endometrial polyps were investigated in 961 patients with abnormal uterine bleeding who underwent dilatation and curettage (D&C) between January and December 2006. Statistical analysis was performed. Results: Endometrial polyps were found in 211 (21.94%) patients with abnormal uterine bleeding. Histopatologically, 175 (82.93%) polyps were benign, 29 (13.74%) polyps had simple and complex hyperplasia, 5 (2.37%) polyps had atipical hyperplasia, and 2 (0.95%) polyps had endometrial carcinoma. According to the malignant potential, endometrial polyps were divided into a group of benign (benign polyps and polyps with simple and complex hyperplasia) and a group of premalignant and malignant (polyps with atypical hyperplasia or endometrial carcinoma). Group of premalignant and malignant polyps were associated significantly with older age, postmenopause and hypertension. Conclusions. The prevalence of endometrial polyps in patients with abnormal uterine bleeding according our data was 21.94%. Atypical hyperplasia and endometrial carcinoma were rarely confined to a polyp. Older age, postmenopause and hypertension may incrise the risk of premalignant and malignant changes in endometrial polyps. PP4-49 THE PROGNOSTIC IMPORTANCE OF GLUT 1 AND KI-67 EXPRESSION IN OVARIAN EPITHELIAL TUMOURS Muberra Segmen Yilmaz, Dilek Yavuzer, Nimet Karadayi, Aylin Ege Gul Dr. Lutfi Kirdar Kartal Education and Research Hospital, Department of Pathology, Istanbul, Turkey BACKGROUND: In epithelial malignancies, aberran expression of glucose transporter 1 (Glut 1), a glucose transport protein located at cell surface and its relation with neoplastic progression is well known. Beside this, protein derived Ki-67 antigen which is observed during proliferative cell cycle indicates proliferation of tumour cells. We studied Glut 1 and Ki-67 expression in benign, borderline, malignant serous and mucinous ovarian tumours, their significance as a prognostic factor in borderline and malignant tumours, their relation with serum Ca 125 levels and other prognostic parameters. METHOD: Ninety two cases (34 serous carcinoma, 4 mucinous carcinoma, 9 borderline serous tumour, 6 borderline mucinous tumour, 19 benign serous cyctadenocarcinoma, 15 benign mucinous cystadenocarcinoma, 5 ovarian endosalphingiosis) in which Glut1 Ab-1 and Ki-67 investigatement of immunohistochemical studies on sections obtained from paraffin blocks were diagnosed at Dr. Lutfi Kırdar Kartal Education and Research Hospital. Patients were analysed according to age, tumour grade, stage and serum Ca 125 levels. RESULTS: In benign, borderline, malignant serous and mucinous tumours and ovarian endosalphingiosis, regarding Glut 1, Ki-67 expression and serum Ca-125 levels were found to be significantly different (p<0.01). Glut 1 did not show any staining in benign cases and endosalphingiosis while 86.7% of borderline cases and 97.4% of malignant cases revealed positive staining. More than 1% positive nuclear staining was observed with Ki-67 in 35.3% of benign cases, 80% of borderline cases and 89.5% of malignant cases. Moreover, Glut 1 staining degree with serous

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tumours was found significantly higher than mucinous tumours (p<0.05). Malignant and borderline cases were significantly correlated with each other due to their Ki-67 and Glut 1 staining degrees (p<0.05). Also, statistically significant correlation was found between Glut 1 staining degree with tumour stage and tumour grade (p<0.05). Ca 125 levels of serous tumours were found higher than musinous tumours (p<0.05). Invasive tumours had higher Glut 1, Ki-67 staining degrees and mean serum Ca 125 levels than borderline tumours (p<0.05). CONCLUSION: This study revealed that Glut 1 expression in serous and mucinous ovarian tumours is related with malignancy progression and other prognostic parameters. Although Ki-67 expression is related with malignancy progression, any other significant relation is not found with other prognostic parameters of malignant tumours. PP4-50 A TEN YEAR REVIEW OF THE PATHOLOGY OF IMAGE-GUIDED PERITONEAL CORE BIOPSIES IN WOMEN PRESENTING WITH PERITONEAL CARCINOMATOSIS Sally Osborn1, Matthew Hewitt2, Michael Weston3, Tim Perren4, Geoffrey Lane5, John Spencer3, Nafisa Wilkinson6 1 Dept of Histopathology St James's University Hospital, Leeds, United Kingdom 2 Dept of Obstetrics and Gynaecology UCC, BUPA Ireland Research Centre, Cork University Maternity Hospital, Cork, Ireland 3 Dept of Clinical Radiology, St James's University Hospital, Leeds, United Kingdom 4 Cancer Research UK Clinical Centre, St James's University Hospital, Leeds, United Kingdom 5 Dept of Gynaecology, St James's University Hospital, Leeds, United Kingdom 6 Dept of Histopathology, St James's University Hospital, Leeds, United Kingdom BACKGROUND: The commonest cause of peritoneal carcinomatosis in women is ovarian or primary peritoneal carcinoma. Presentation in the vast majority of women is with disseminated abdominal disease. Surgery in advanced cases is unlikely to achieve optimal debulking in some patients and therefore, primary neoadjuvant chemotherapy with the aim of achieving substantial cytoreduction to be followed by interval debulking surgery is a management option. Other patients with advanced disease are too ill for major cytoreductive surgery and chemotherapy is the treatment of choice. Therefore, an accurate histological diagnosis to establish the primary site of origin is paramount for the management of these patients. Image-guided peritoneal biopsy is a simple and safe technique to provide sufficient tissue in the majority of cases to reach an accurate diagnosis. METHOD: A series of 201 women who presented to a teaching hospital with peritoneal carcinomatosis over a ten year period underwent image-guided (abdominal ultrasound or computer tomography) peritoneal core biopsy, following multidisciplinary team review. Multiple cores were obtained and examined with haematoxylin and eosin. Ancillary techniques were used where indicated by the clinical situation; this was particularly helpful in the distinction of endometrioid adenocarcinoma from metastatic colorectal carcinoma. The slides of any known previous malignancies were reviewed and compared with the current biopsy to exclude recurrence. RESULTS: A primary site of origin was identified in 188 women; of these 164 were mullerian in origin, the majority of which were serous papillary carcinoma (130). The remainder were endometrioid 7, mucinous 4, clear cell 3, squamous cell carcinoma 1, mixed endometrioid and serous 2, carcinosarcoma 7, poorly differentiated mullerian carcinoma 10. 24 originated from other sites (gastrointestinal tract 9, breast 6, lymphoma 3, pseudomyxoma 2, hepatobiliary 2, renal 2). In 7 women the biopsy showed poorly differentiated carcinoma with no primary

site identified and in 5 women the biopsies showed benign changes. In 1 patient the initial biopsy led to a misdiagnosis and a laparoscopic biopsy was required to make the definitive diagnosis. In 10 women there was insufficient material on the initial biopsy to make a diagnosis, a final diagnosis was made on repeat biopsy. CONCLUSION: In conclusion, peritoneal core biopsy in a multidisciplinary setting, together with appropriate biochemical markers, is a quick, efficient and safe method of making a reliable, definitive diagnosis in the vast majority of women with peritoneal carcinomatosis. PP4-52 EXPRESSION OF MATRIX METALLOPROTEINASE-2 AND ITS TISSUE INHIBITOR IN ASSOCIATION WITH CD44V6 IN THE NEOPLASTIC SEQUENCE OF SEROUS OVARIAN TUMOURS Duygu Enneli Kankaya1, Ebru Erol1, Arzu Ensari1, Yasemin Genc2, Ayse Sertcelik1 1 Ankara University, School of Medicine, Department of Pathology, Ankara, Turkey 2 Ankara University, School of Medicine, Department of Biostatistics, Ankara, Turkey Background: Variants of CD44, a cell surface adhesion molecule, have been suggested to be important in cancer invasion and metastasis and proposed as a potential prognostic marker in many epithelial and non-epithelial malignancies. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are also known to play a role in invasion and metastasis of many tumours. There have been controversial results regarding the significance of MMPs, TIMPs and CD44v6 for the development and progression of ovarian tumors. Therefore, the aim of the present study was to investigate the expression of CD44v6, MMP-2, and TIMP-2 in ovarian serous neoplasms and to evaluate their clinical significance. Methods: A total of 56 cases, which include 9 serous cysts, 10 serous cystadenofibromas, 3 borderline serous tumours and 34 serous carcinomas was included in the study. Clinical parameters such as age of the patient, tumour size, grade, stage were retrieved from the patient files. Tissue microarray blocks were constructed by taking 3 cores 2 mm in diameter for each case and immunohistochemistry was performed for the evaluation of the expression of CD44v6, MMP-2, TIMP-2 in sections of 3 m thick. Expression of CD44v6 was evaluated by scoring intensity and percentage of staining, while expression of TIMP-2 and MMP-2 were evaluated only by scoring staining intensity on an arbitrary grading scheme of (+)-(+++). Kruskal- Wallis and Pothoc test were used for statistical analysis. Results: Both MMP-2 and TIMP-2 were expressed by the fibroblastic cells in ovarian stroma as well as vascular endothelium as well as the epithelium while CD44v6 was only expressed in the epithelial tissue. No difference between stromal and epithelial expressions of MMP-2 and TIMP-2 in any of the study groups was observed.The expression of TIMP-2 and MMP-2 were significantly higher in borderline and malignant group than benign group (p<0,05), while no such difference was observed between borderline and malignant cases. TIMP-2 expression was higher in serous cystadenofibromas compared with serous cyst (p<0,05). There was no significant difference in the CD44v6 expression between the study groups. No correlation was found between the expressions of all three markers and the clinical features of the patients. Conclusion: MMP-2 and TIMP-2 expressions seem to correlate with the neoplastic sequence of serous ovarian neoplasms whereas no such correlation was observed in CD44v6 expression. These findings suggest that there is increased degradation of extracellular matrix by MMPs in parallel with neoplastic progression.

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Kidney Pathology PP4-53 NEPHROBLASTOMA WITH BONE METASTASIS IN A 30-YEAR WOMAN, A CASE REPORT Amel Trabelsi, Soumaya Rammeh, Sarra Mestiri, Yacoubi Med. Tahar, Sihem Hmissa, Moncef Mokni, Korbi Sadok Pathology Department, F. Hached Hospital, 4000 Sousse, Tunisia Background: Nephroblastoma is rare in adults and thought to have a worse prognosis than its counterpart seen in children. Bone metastasis of nephroblastoma are very rare in the 2 age groups. Method: We report a case of 30 year-old woman, that consulted for abdominal distension associated to pain in the left hip. Results: Imaging findings showed a right renal tumor measuring 18 cm with a condensing lesion of the superior extremity of the left iliac bone. The patient underwent a right nephrectomy and biopsy of the left iliac bone. Histologically, the renal tumor was a biphasic nephroblastoma without cellular anaplasia. The bone biopsy showed a round cell proliferation similar to the blastema cells seen in the renal tumor. The patient had 3 courses of chemotherapy without clinical improvement then has been out of sight. Conclusion: Nephroblastoma in adult with bone metastasis is very rare with a poor prognosis. PP4-54 PAPILLARY CARCINOMA OF THE KIDNEY: A RETROSPECTIVE STUDY OF 6 CASES Mestiri Sarra, Trabelsi Amel, Ghaieb Amel, Ben Yacoub-Abid Lilia, Aachech Thouraya, Rammeh Soumaya, Mokni Moncef, Korbi Sadok Pathology Department, F. Hached Hospital, Tunisia Background: Papillary carcinomas of the kidney are relatively rare tumors, counting for 4% of renal cancers. Two types are actually recognized, that differ by their morphological and prognostic features. Method: In a retrospective study, we have studied the clinicopathological and imaging features of 6 cases of papillary carcinoma retrieved in an 11- year period (1995-2005) at the pathology department of F. Hached hospital. These tumors were graded according to the new 2004’s OMS histopathological classification of renal tumors. Results: In our study, there were 5 men and one woman. The age median was of 68 years with extremes ranging between 57 and 83 years. The tumors were asymptomatic in 3 cases, they were revealed by lumbar pain in 3 cases. A lombar contact and a lombar sensibility were found in 2 cases. Imaging findings were various, but ultrasonography and CT-scan found a neoplastic process in 5 cases. Histopathological examination found that type 2 papillary carcinoma was correlated to a higher nuclear grade, to an advanced infiltration stage and to a poorly prognosis. Treatment consisted in a radical nephrectomy in 5 cases and in a tumorectomy in one case. Conclusion: Actually, it is important to classify papillary carcinoma into type 1 or 2 because of the prognostic implications. PP4-55 THE MORTALITY AND CLINICALLY MISSED DIAGNOSIS OF AA AMYLOIDOSIS IN RHEUMATOID ARTHRITIS Miklós Bély1, Ágnes Apáthy2 1 Policlinic of the Hospitaller Brothers of St. John of God in Budapest, Hungary 2 National Institute of Rheumatology and Physiotherapy, Budapest, Hungary Introduction AA amyloidosis (AAa) is one of the most important complications of rheumatoid arthritis (RA). Aim of this study was to determine: 1. the mortality due to AAa in RA, 2. the clinically missed diagnosis of AAa in RA, 3. the correlations between severity, mortality, and clinical recognition of AAa.

Patients and Methods A randomized (non-selected) autopsy population of 161 in-patients with RA was studied (diagnosed clinically according to the criteria of the American College of Rheumatology-ACR). The basic disease, the complication(s), and the cause of death were determined and analyzed retrospectively, reviewing the clinical and pathological reports, tissue samples, and the histological slides. Amyloidosis and severity of amyloidosis was determined histologically. The correlations were determined by ²-test, comparing the mortality and clinically missed diagnosis of AAa with mild (<) amyloidosis. Results 1), or severe (1 1. Systemic secondary (AA) amyloidosis was observed in 34 (21.1%) of 161 patients. Thirteen (38.2 rel%) of 34 patients revealed a “severe” degree of amyloid A deposition, massively involving many tissue structures of examined ); and 21 (61.8 organs (with average amount of amyloid A deposits / patient 1 rel%) cases were regarded as “mild”, involving only a few tissue structures in some organs (with average amount of amyloid A deposits / patient <1). 2. Seventeen (50 rel%) of 34 patients died of uremia, and 9 (26 rel%) of 17 were diagnosed clinically. 3. There was a significant and positive correlation between severity and mortality of AAa ( ²=7.97, p<0.004), or clinical recognition of AAa ( ²=5.98, p<0.01), furthermore between mortality and clinical recognition of AAa ( ²=9.67, p<0.001). Conclusions AA amyloidosis should be regarded as one of the most insidious complications of rheumatoid arthritis (RA), which may furtively lead to death (Bély, 1993). The clinical recognition of AAa is quite effective. In half of the lethal cases amyloidosis has been recognized clinically, but all of these were in a late, advanced stage of amyloidosis, with massive renal amyloid deposits and uraemia. The early diagnosis of AAa is important, because early recognition of amyloidosis allows an early start of therapy and therefore entails a better prognosis. Biopsy is suggested in all hospitalised RA patients, with or without clinical signs of amyloidosis. PP4-56 STAGE DEPENDENT QUANTITATIVE DIFFERENCES OF (AA) AMYLOIDOSIS IN RHEUMATOID ARTHRITIS Miklós Bély1, Ágnes Apáthy2 1 Policlinic of the Hospitaller Brothers of St. John of God in Budapest, Hungary 2 National Institute of Rheumatology and Physiotherapy, Budapest, Hungary Introduction Development of systemic AA amyloidosis (AAa) is a progressive, cumulative process. Aim of this study was to determine 1. the quantitative differences of amyloid A deposits in RA patients complicated with AAa at death, 2. the quantitative differences of amyloid A deposits in different tissue structures of various organs. Patients and Methods A randomized (non-selected) autopsy population of 161 in-patients with RA was studied. RA was diagnosed clinically according to the criteria of the American College of Rheumatology. AAa was detected histologically. The quantitative differences of amyloid A deposits on different structures of various organs at death were evaluated by semi-quantitative, visual estimation of amyloid deposits on a 0 to 3 plus scale. Results AAa was observed in 34 (21.1%) of 161 patients. Thirteen (38.2 rel%) of 34 patients revealed a “severe” degree of amyloid A deposition, massively involving many tissue structures of examined organs (with average amount of ); and 21 (61.8 rel%) cases were regarded as amyloid A deposits / patient 1 “mild”, involving only a few tissue structures in some organs (with average amount of amyloid A deposits / patient <1). Discussion The process of amyloid A deposition is a progressive and time dependent phenomenon. Minimal or moderate amyloid A deposits (with average amounts of amyloid A deposits / patient <1), involving only a few tissue structures in some organs representing the early stage of amyloidosis. Massive amyloid A deposits (with average amounts of), massively involving many tissue structures of amyloid A deposits / patient 1 examined organs represents the late (advanced) stage of amyloidosis.

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During the progression of amyloidosis, amyloid A protein deposition begins in the organs and tissue structures that are frequently involved and later show marked deposition of amyloid. Where deposits are infrequent or less marked, deposition starts later. Amyloid deposition in the wall of blood vessels (capillaries, arterioles, small arteries) will be followed by deposition of amyloid A along collagen fibers, basal membranes, and reticulin fibers (fat tissues). The amyloidosis of peripheral nerves is a late, end stage phenomenon. The chronologic sequence of amyloid A deposition regarding the organs is: GI tract, heart, kidney, spleen, liver and the adrenal glands. Other organs like lungs, pancreas, thyroid gland, aorta, skeletal muscle, synovial membrane, lymph nodes, peripheral nerves, bones and skin are involved less frequently and amyloid A deposition starts later in them. PP4-57 NEPHROTIC SYNDROME AND ACUTE RENAL FAILURE IN NON-HODGKIN LYMPHOPLASMOCYTIC LYMPHOMA WITH MONOCLONAL GAMMOPATHY Tatjana Terzic1, Natasa Colovic2, Milica Colovic2, Bosko Andjelic2, Biljana Mihaljevic2, Jasmina Markovic-Lipkovski1 1 University of Belgrade, Faculty of Medicine, Institute of Pathology, Serbia 2 University Clinical Centre of Serbia, Institute of Hematology, Serbia Background. Lymphoplasmacytic lymphoma is a rare disease that occurs in older adults, characterized by proliferation of small B lymphocytes, plasmacytoid lymphocytes or plasma cells with or without production of serum monoclonal protein. Renal lesions may occur quite rarely and may be the result of amyloidosis or depositing of monoclonal immunoglobulins or their subunits in one or more renal compartments including glomeruli, tubules, interstitium and blood vessels. Case report. We report two patients with lymphoplasmacytic lymphoma and monoclonal gammopathy of IgM and IgG type, nephrotic syndrome and acute renal failure. A 58-year-old man previously treated for pre-B acute lymphoblastic leukemia, developed 3 years later nephrotic syndrome as a complication of lymphoplasmacytic lymphoma and high paraprotein IgM kappa type. Biopsy of the kidney with light microscopic examination revealed 12 glomeruli with increase of mesangial matrix and mesangial cells. This proliferation was of segmental character with nodular appearance. Glomerular basal membrane was thickened and duplicated. Five glomeruli showed capsular adhesions. There were moderate tubular atrophy and interstitial fibrosis. The small arteries showed hyalinization of the walls. Imunofluorescence revealed mesangial and capillary wall positivity for IgM and kappa light chains and negativity for lambda light chains. Diagnosis of membranoproliferative glomerulonephitis was made. Treatment with cyclophosphamide was ineffective and patient died 2 months later. The second patient is 42-year-old female diagnosed with lymphoplasmacytic lymphoma and paraprotein IgG lambda type. Kidney biopsy disclosed 9 glomeruli, among them one was completely sclerotic. Glomeruli showed hypercellularity due to mesangial cell proliferation and infiltration with mononuclear leukocytes. There were moderate to severe tubular atrophy and interstitial fibrosis. Few tubules were extremely dilated filled with amorphous eosinophilic material with impression of microcystic formations. There were patchy interstitial infiltrates with mononuclear leukocytes. Immunofluorescence staining of glomeruli revealed mesangial and tubular wall positivity for immunoglobulin G, component of complement C3 and lambda light chains. Haemodyalisis and cytostatic therapy were without response and she died after 45 days. Conclusion. In our both cases light chain deposits were found. These light chains have probably caused glomerular as well as tubular changes which were observed.

PP4-58 RENAL MEDULLARY CARCINOMA IN A CHILD WITHOUT SICKLE CELL DISEASE: CASE REPORT Lina Gomes Dos Santos1, Aracy Carstens Cunha2, Benedito Borges Da Silva3, Jerusia Oliveira Ibiapina4, Teresinha Castelo Branco Carvalho4, Kelson James Almeida3, Gildene Alves Da Costa4, Daniel José Martins Barbosa4, Euripedes Soares Filho4, Jucelia Saraiva E Silva5, Ana Maria Lima Furtado Veloso4, Joao Bosco Parentes Vieira3 1 Federal University of Piauí, Brazil 2 Diagnose Cunha Lab, Brazil 3 Federal University of Piauí, Brazil 4 São Marcos Hospital, Brazil 5 Estadual University of Piauí, Brazil Background: Renal Medullary Carcinoma is a rare primary neoplasm of the kidney more commom in young adults and frequently associated to sickle cell disorders. It was described by Davis et al in 1995 as epithelial tumor arising from the renal calix of patients with sickle cell disorders. Case Report: A 15-year-old black child admitted with flank pain lasting one year, hematuria and weight loss. Examination showed a palpable mass in the left flank. Laboratory tests were performed and showed: hematocrit 25%, leukocytes 11,400mm3, normal values of electrolytes and renal/hepatic function parameters. Hemoglobin electrophoresis and falciform anemy test were negative. Computed tomography of abdome showed a centrally located mass in left kidney. Then, radical nephrectomy was performed and the pathological result was a well-circumscribed tumor with 10,0 x 8,0cm located in kidney medulla. Microscopically, the neoplasm consisted of solid sheets of cells, eccentric nuclei, prominent nucleoli and eosinophilic and granulated abundant cytoplasm with dense infiltrate of lymphocytes and polymorphonuclear leukocytes. Immunohistochemical stains performed were positive for vimentine, CK7 and CK8 and negative for CK20 and CD10. Despite of treatment stablished the patient died within twelve months. Conclusion: Renal Medullary Carcinoma, known as seventh sickle cell nephropathy, a rare and extremmelly aggressive renal tumor, leading to the death of the majority of patients, as happened in the present case. PP4-59 RENAL INFLAMMATORY PSEUDOTUMOR: A CLINICOPATHOLOGIC AND IMMUNOHISTOCHEMICAL STUDY OF A NEW CASE Petrescu Amelia Nicoleta1, Berdan Gabriela1, Ambert Valentin1, Jinga Viorel1, Popescu Mircea1, Andrei Florin2, Hulea Ionela3, Gaitanidis Raluca3, Niculescu Liviu4 1 Th. Burghele” Hospital , Bucharest, Romania 2 Victor Babes” Institute of Pathology, Bucharest, Romania 3 Th. Burghele” Hospital, Bucharest, Romania 4 University of Medicine and Pharmacy “ Carol Davila”, Bucharest, Romania Renal inflammatory pseudotumor is an uncommon benign tumor that has been classified into separate group, but there is a risk that this lesion could be misdiagnosed. The aim of this work is to report a new case of 57 year old man presented in our hospital with hematuria, minimal grade fever and right flank pain. Magnetic resonance imaging ( MRI) and sonography revealed a tumor of the right mediorenal parenchyma, 2,5 cm in diameter. The patient underwent right nephroureterectomy under the diagnosis of renal cell carcinoma. Macroscopically examination carried out the removed kidney showed a 2/2/1,5 cm yellowish, gelatinous, well circumscribed, mediorenal and pericaliceal mass. Material and methods: Fragments of the tumor were fixed in formaldehyde 10%, included in paraffin and the sections were stained with HE,VG and immunohistochemically: vimentine (VIM), MNF116, SyN, smooth muscle actin ( ACT), DESMINE, CD68, S100. Results: The histological examination revealed a compact spindle cell proliferation, a hypocellular fibrous area in a

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oedematous myxoid background infiltrated by small lymphocytes, histiocytes ,some plasma cells and small bone area. The spindle cells were diffuse positive for VIM, ACT, CD 68 and negative for desmin, MNF 116, SyN and S 100. Conclusions: The pathologic diagnosis was renal inflammatory pseudotumor and raises the problem of differential diagnosis as the clinical and imagistic aspects are similar to those of a renal carcinoma and the problem in establishing a preoperative correct diagnosis. PP4-60 PAPILLARY RENAL CARCINOMA – A MORPHOLOGIC STUDY OF 36 CASES Monica Gratiela Hortopan Fundeni Clinical Institute Bucharest, Romania ABSTRACT: The study consisted of 36 patients diagnosed and surgically treated in the Center of Urological Surgery, Dialysis and Renal Transplant from Fundeni Clinical Institute between January 2002 and December 2006. We have evaluated the histological spectrum of PRCCs and assessed significance of conventional pathologic prognostic parameters (Fuhrman’s nuclear grade, pathologic stage, tumour size, multifocality, necrosis, and foam cells) and correlated these with outcome. The conclusion that we reached from our study are: 1. PRCC is a malignant tumour, diagnosed at a lower stage, with a distinct potential for progression and aggressive behaviour. 2. Evaluation of these tumours according to cell type, amount of foam cells, presence or absence of vascular invasion, nuclear grade, and pathologic stage provides useful prognostic information. 3. The better 5-year survival rate of papillary RCC (overall and for stage I tumours) compared with that of conventional RCC suggests that it is a tumour with lower malignant potential. 4. Differential diagnosis must be made with urothelial carcinomas, collecting duct carcinomas, metastatic papillary carcinomas (lung, thyroid, etc.)and clear cell carcinoma. PP4-61 NESTIN EXPRESSION IN MEMBRANOUS GLOMERULONEPHRITIS Jasmina Markovi -Lipkovski1, Ljiljana Gojkovi -Bukarica2, Sanja itlu anin1, Radmila Stevanovi 1, Sanja Radojevi -Škodri 1, Gordana Basta-Jovanovi 1, Biljana Stojmirovi 3, Vida Neši 3 1 Institute of Pathology, School of Medicine, University of Belgrade, Serbia 2 Institute of Clinical Pharmacology, Pharmacology and Toxicology, School of Medicine, University of Belgrade, Serbia 3 Institute of Nephrology and Urology, Clinical Centre of Serbia, Belgrade, Serbia Background. Nestin, an intermediate filament protein which has role in regulating cellular cytoskeletal structure, is restrictedly expressed in the podocytes of human adult kidneys. In the present study nestin expression was investigated in biopsy specimens of membranous glomerulonephritis (MGN). Method. During the last two years, 52 MGN were collected out of 534 kidney biopsy received to be diagnosed at our institution. The diagnoses were performed by light and immunofluorescent microscopy and in some cases by electron microscopy. Clinical data (including immunological analysis wiht ANA and antiDNA antibodies) were also taken into account. For immunomorphology, monoclonal anti-nestin antibody from mouse (SC-23927, clone 2C1.3A11, Santa Cruz Biotechnology) diluted 1:100 was applied on cryostat or paraffin sections using Labeled StreptAvidineBiotin (LSAB+ Dako®) method. Visualization was performed by Dako® AEC substrate. Ten kidney biopsies of patients without nephrotic syndrome, mainly with mesangioproliferative GN, were used for control staining. Results. 52 patients were middle-aged adults (mean age 46) and 4 patients were under age of 15, ratio female : male was 26 : 30. In idiopathic MGN routine immunofluorescence analysis revealed

granular deposits of IgG and C3 along glomerular basement membrane (GBM), and by light mtcroscopy the glomeruli showed thickened basement membrane, but appeared normocellular. Mesangial proliferation in addition to uniform thickening of GBM was crucial finding for secondary MGN and immunofluorescence microscopy usually showed a ”full house” pattern (fine granular deposits of IgA, IgG, IgM, C1 and C3 along the GBM). In 12 cases (21.4%) idiopathic MGN was diagnosed, and in 35 cases (62.5%) secondary MGN. In 9 cases (16.1%) etiology was undetermined. Nestin expression was variably present in glomeruli in different cases of MGN. Decreased expression was predominantly found in MGN of undetermined or secondary etiology. Conclusion. MGN revealed heterogenity concerning nestin expression. Nestin expression was especially diminished in MGN associated with so-called ”full house” immunofluorescence findings. PP4-62 SEVERAL PATHOGENIC MECHANISMS OF CONGENITAL GLOMERULOPATHIES Maida Tussupbekova Karaganda State Medical Academy, Kazakhistan At present time, problems of kidney pathology of fetus and newborn at complicated pregnancy are not completely investigated. Different factors cause delay of fetus and newborn tissues maturity from their hestational age with persistence of embrional structures, became a cause of congenital developmental defects formation and make difficult adaptation of the organism to extra uterine life. Aim of the research: to study the structure of mortality and pathological morphology of kidneys of fetus and newborns at complicated pregnancy. The analysis 566 still-born children and 516 newborns, dead in perinatal periodwas done. There were allocated groups of research - control group, with anemia, and chronic pyelonephrotis. Object of research was both kidneys. Common histological methods, immune fluorescent investigation and morphometric research were made. Results of the research: The basic cause of death is asphyxia (35,6±2,3% against 54,4±1,9%),second place have respiratory disorders of lungs. Congenital developmental defects are on third place, their frequency is increased in two times till 17,6±1,8%, against 7,93±1,0%. Increase of cases of intra uterine infection till 16,7±1,8%, against 3,6±0,7% is also visible. Morphometric criteria of hestational and pathological immaturity of kidneys: number of layers and volumetric part of renal flomeruli, number of embryonic nephrons, and square of tubular surface. Hestosis of pregnant women is a high risk factor of renal dysembriogenesis. Pathological immaturity of kidney tissue at normal duration pregnancy and hestosis is diagnosed at 74,0±5% cases, focal and cystic cortical dysplasia of kidney structure was revealed at 13,0±4%. At histological research of kidneys of fetus and newborns in group with hestosis of pregnant women in 11,0±3% membranous and membranous –proliferative variants of glomerulopathies was revealed. At immune fluorescent investigation bright granular glow of IgM on basal membranes with blade changes of vascular glomeruli and line glow of C3 component of compliment were marked, these are testify about possible participation of anti renal immune factors of pregnant women in the development of congenital immune complex glomerulopathy. In the group of investigation pathological immaturity was revealed in 38,7±5,3% at presence of chronic pyelonephritis at mothers. Analysis of data of pathological morphological research of kidneys of fetus and newborns at complicated pregnancy describes some pathogenic mechanisms of congenital glomerulopathies development and causes, leading nephrogenesis disorders.

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PP4-63 THE INFLUENCE OF TUBULAR PHENOTYPIC CHANGES ON THE DEVELOPMENT OF DIFFUSE INTERSTITIAL FIBROSIS IN RENAL ALLOGRAFTS B. Handan Ozdemir, Nihan Haberal, F. Nurhan Ozdemir, Beyhan Demirhan, Mehmet Haberal Baskent University, Faculty of Medicine, Turkey It has been reported that myofibroblasts are the major cells in the development of interstitial fibrosis (IF) and therefore chronic graft dysfunction in renal allografts. In normal human kidney, tubular cells do not have myofibroblast differentiation and they don’t have alpha-smooth muscle actin (alpha-SMA) expression. In this study we aimed to show that tubular cells can undergo phenotypic changes toward myofibroblasts and induce early IF and poor graft outcome in renal allografts. The expression of alpha-SMA and the formation of vinculin and paxillin containing adhesion complexes are the primary criteria for determining the differentiation of non-muscle cells such as renal tubule cells into contractile myofibroblasts. For this reason we immunostained first year renal allograft biopsies of 74 patients with alpha-SMA, Vinculin and Paxillin primary antibodies and the expression of tubules and glomerular cells were evaluated. Myofibroblast differentiation of renal tubules (alpha-SMA, vinculin and paxillin positive tubules) was found only 30 of 74 patients. In addition glomerular cells of 36 patients showed positive alpha-SMA, vinculin and paxillin staining. The development of diffuse IF was found significantly early in cases with tubules showing myofibroblast differentiation compared to cases with tubules that did not have myofibroblast differentiation (p<0.01). The presence of proteinuria in first year showed significant positive correlation with the glomerular alpha-SMA, vinculin and paxillin staining (p<0.001). Cases whom showed tubular and glomerular alpha-SMA, vinculin and paxillin staining showed worse graft outcome compared to cases that did not show tubular and glomerular staining (p<0.001). In conclusion our results showed that renal tubular and glomerular cells can show myofibroblastic differentiation and these cells have a role in the development of diffuse interstitial fibrosis and early proteinuria in renal allografts. PP4-64 PERITUBULAR CAPILLARY AND VASCULAR MACROPHAGE INFILTRATION CORRELATES WITH MICROVASCULAR DESTRUCTION AND WORSENS STEORID RESPONSE AND RENAL ALLOGRAFT OUTCOMES FOLLOWING C4d NEGATIVE ACUTE REJECTION EPISODES B. Handan Ozdemir, Nihan Haberal, Feza Karakayali, Beyhan Demirhan, F. Nurhan Ozdemir, Mehmet Haberal Baskent University, Faculty of Medicine, Turkey We aimed to understand the influence of peritubular capillary (PTC) and vascular macrophage infiltration on steroid response and renal allograft outcomes after acute rejection (AR) episodes. Seventy-nine patients with biopsy-proven AR in their first year after transplantation were included in the study. Thirty patients with normal first year renal allograft biopsies were also included in the study and used as a control group. All biopsies were C4d negative. Immunohistochemically we assessed the degree of macrophages (CD68) and HLA-DR-positive infiltrating cells in PTC’s, glomeruli, and on vascular walls and tubules. In addition HLA-DR expression of PTC’s was also evaluated. The decreasing intensity of peritubular capillary HLA-DR (PTC-DR) expression was accepted as the increasing degree of the destruction of PTC. Compared to control group AR cases showed significantly higher degree of macrophage and HLA-DR positive inflammatory cell infiltration in PTC’s, glomeruli, and on vascular walls and tubules (P<0.001 for all). PTC destruction was significantly higher in AR cases than control group (p<0.001). PTC, glomerular and vascular macrophage infiltration showed

significant correlation with PTC destruction and steroid response (p<0.001 for all). Severity of PTC destruction with accompanying higher degrees of macrophage infiltration in PTC’s, glomeruli and on vascular walls caused unresponsiveness to steroid therapy (p<0.001) and poor graft outcome (p<0.001). Five-year graft survival was 95%, 37% and 22% for cases with grade 0, 1 and 2 PTC macrophage infiltration respectively (P<0.001). In addition five-year survival was 80%, 36% and 1% for cases with grade 0, 1 and 2 vascular macrophage infiltration respectively (p<0.001). In conclusion peritubular capillary and vascular macrophage infiltration are important predictors of steroid response and renal outcome following acute rejection in cases whom especially had negative C4d. PP4-65 ROLE OF POLY (ADP-RIBOSE) POLYMERASE ON MICROVASCULAR INJURY AND INFLAMMATION IN RENAL ALLOGRAFT REJECTION AND ITS INFLUENCE ON RENAL GRAFT SURVIVAL B. Handan Ozdemir, Turan Colak, Feza Karakayali, Beyhan Demirhan, Mehmet Haberal Baskent University, Faculty of Medicine, Turkey Introduction: The activation of poly (ADP-Ribose) polymerase (PARP) is well considered to play an augmenting role in inflammation and cell death. The aims of this study were to investigate the role of PARP in acute rejection (AR) and to assess the influence of PARP on renal survival. Methods: Study compromised 81cases and 55 of them had AR. Twenty-six cases had no pathology and used as a control group. PARP and HLA-DR expression of tubules, interstitium, arteries and peritubular capillaries (PTC’s) were studied immunohistochemically and CD68 positive macrophage infiltration of tubules, interstitium, PTC’s and arterial walls were evaluated. The decreasing intensity of PTC HLA-DR (PTC-DR) expression was accepted as the increasing degree of the destruction of PTC’s. Results: AR cases showed higher degrees of tubular, interstitial and vascular PARP and HLA-DR expression compared to control group (p<0.01 for all). PTC-DR expression was lower and PTC-PARP expression was higher in AR cases compared to control group (p<0.001). Increasing of AR grade with the high level of PTC-PARP expression, caused decrease of PTC-DR expression and increase of PTC destruction (p<0.01). Tubular and interstitial HLA-DR expression, interstitial, tubular, vascular and PTC macrophage infiltration showed positive correlation with tubular, interstitial, PTC and vascular PARP expression (p<0.01 for all). In contrast PTC-DR expression showed negative correlation with all these parameters (p<0.01). Severity of PTC destruction with accompanying higher degrees of PARP expression on tubules, interstitium, arteries and PTC’s caused unresponsiveness of steroid therapy (p<0.01) and poor graft outcome (p<0.01). Conclusion: Increased PARP activation leads to higher degrees of cell death and inflammation that AR cases with high renal PARP expression showed significant PTC destruction and renal inflammation. Therefore we suggest that PARP inhibitor drugs can combine with immunosuppressive therapy in order to control PTC destruction and renal inflammation PP4-66 EFFECTS OF SULPHITES ON KIDNEY HISTOLOGY IN YOUNG AND ELDERLY RATS Nagihan Yalcin1, Murat Colakoglu2, Vural Kucukatay3, Erdogan Kocamaz4 1 Department of Pathology, Pamukkale University, School of Medicine, Denizli, Turkey 2 Department of Nephrology, Pamukkale University, School of Medicine, Denizli, Turkey 3 Department of Physiology, Pamukkale University, School of Medicine, Denizli, Turkey 4 Department of Histology and Embrylogy, Pamukkale University, School of Medicine, Denizli, Turkey

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BACKGROUND:The human body is constantly generated from the metabolism of the sulfur containing amino acids. Sodium metabisulphite is used as an antioxidant and antimicrobial agent in a variety of drugs and functions as preservative in many food preparations. There is no knowledge about the histological effects of sulphites on kidneys. The aim of this study was to investigate the possible toxic effects of sulphites on kidneys by assessing histological findings in young and elderly rats. MATERIALS AND METHODS: Rats were assigned to four groups: Two sulphite administrated young and elderly groups, and two young and elderly control groups. Sulphite was administrated in the form of sodium metabisulphite 70mg/kg/day via drinking water. The kidney tissues were fixed on buffer formalin and then they were stained with H&E, methenamine silver, PAS and Masson’s thricrome. RESULTS:There were no glomerular changes (i.e. thickness of basement membrane, focal segmental sclerosis), tubular changes, tubular interstitial changes or vascular changes. No differences between young and elderly sulphite administrated groups were observed histologically. CONCLUSIONS:In this study, the specimens were examined by light microscopy. Further studies done by electron microscopy are needed to support these findings. PP4-67 HISTOPATHOLOGICAL STRUCTURE OF THE KIDNEY IN HIGH FAT DIET FED RATS: A LIGHT MICROSCOPICAL STUDY Deniz Unal, B. Zuhal Altunkaynak, M. Eyup Altunkaynak Department of Histology and Embryology, School of Medicine, Ataturk University, Erzurum, Turkey Background: To characterize kidney in high fat induced obesity model we have examined renal structure in adult Sprague-Dawley rats fed a control diet (CD) or fat-rich diet (HFD) during three months. Methods: Ten adult female Sprague Dawley rats were fed a diet constituted highly of fat (%30) for duration of 3 months. Ten control rats were maintained with a standard rat chow. All animals were weighed per 10-day for 3 months. At the end of the experiment, the naso-anal length of the anesthetized rats was measured to calculate the body mass index (BMI), and subsequently, whole kidneys of the intracardially formalin-perfused animals were removed. Qualitative features of kidney were examined histologically. Results: Light microscopic investigation showed a dilatation in blood vessels and bowman capsule, mononuclear cell infiltration, degeneration in nephrons including glomerulosclerosis and tubular defects and an increase in the connective tissue in the kidneys from the treatment group. Conclusion: Eventually, we have thought that fatty diet causes obesity and may lead to renal failure as a result of histopathological changes. PP4-68 EXPRESSION OF INSULIN-LIKE GROWTH FACTOR IN PROLIFERATIVE GLOMERULONEPHRITIS Fulya Cakalagaoglu1, Zeynep Tosuner1, Naziye Ozkan1, Serhan Tuglular2, Beyza Macunoglu2 1Marmara University Faculty of Medicine, Department of Pathology, Istanbul, Turkey 2Marmara University Faculty of Medicine, Department of Nephrology, Istanbul, Turkey Background Insulin-like growth factor-1 (IGF-1), a peptide growth factor produced by collecting ducts, and its receptor, IGF-1 receptor are present in the glomeruli and basolateral membrane of renal proximal tubular cells. Animal studies showed that IGF restores nephron growth and contributes to tissue growth by causing arterioler dilatation, increasing the glomerular filtration rate and accelerating tubular regeneration.The present study aimed that analyse the expression of IGF-1 in proliferative glomerulonephritis(PGN) and focused on its biologic significance in glomerulonephritis. Methods.IGF -1 expression was studied in

34 non diabetic specimens: normal kidney (6), proliferative glomerulonephritis (GN,14), nonproliferative GN (14). All slides were immunohistochemistry stained with IGF-1(G-17; SC-1422; Santa Cruz Biotechnology; California). In all cases, IGF-1 expression was scored semiquantative by two pathologists ( 0-3 ) in glomeruli and tubulointerstitial. Routine histochemical and immunofloresanse stains were performed and evaluated all cases. Activity index, chronicity, interstitial injury and sclerotic glomeruli were examined in all cases. Results.In normal kidney there was strong IGF-1 immunreactivity in the proximal tubules and minimal IGF -1 expression in the glomeruli.We found increased IGF-1 immunexpression in glomeruli at Proliferative GN groups compared with nonproliferative groups. IGF-1 expression was correlated with the activity index for Proliferative GN groups. Tubulointerstitial IGF-1 expression was similar in proliferative and non proliferative GN groups. We did not found a statistically significant correlation between fibrosis and glomerul IGF-1 expression in two groups. Only significant correlation was between fibrosis and tubulointerstitial IGF-1 expression. Both groups had similar sclerotic glomeruli proportions. Conclusion: These data indicate that IGF-1 indicates the activity in proliferative glomerulonephritis, but larger group studies are needed to confirm our results PP4-69 EFFECT OF FK506 IN EXPERIMENTAL GLOMERULONEPHRITIS: LIGHT MICROSCOPIC AND ULTRASTRUCTURAL STUDY Fulya Cakalagaoglu1, Naziye Ozkan2, Emine Salva3, Serap Arbak4, Bahar Uslu4 1 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department; Marmara University, Medical Faculty, Pathology Department, Istanbul, Turkey 2 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department; Marmara University, Medical Faculty, Pathology Department, Istanbul, Turkey 3 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department, Istanbul, Turkey 4 Marmara University, Medical Faculty, Histology and Embriology Department, Istanbul, Turkey Background The anti-Thy1.1 model is a rat model of mesangial proliferative glomerulonephritis characterized by mesangial cell proliferation and accumulation of mesangial matrix expansion with subsequent resolution and return to almost normal histology. FK506 is an immunosuppresive drug used for treatment of autoimmune disease and after transplantation.In present study, we investigate the effect of FK506 on experimental glomerulonephritis model. Method We studied the expression of

-Smooth Muscle Actin( -SMA), Proliferative Cell Nuclear Antigen(PCNA),Macrophage Calprotectin MAC), aminine, ascular Endothelial Growth Factor(VEGF) in paraffin embedded tissue section from kidney by immunhistochemical method and apoptosis with TUNEL.We studied kidney tissue with fluorescein isothiocyanate(FITC).IgG,IgM,IgA,C3,C1q and fibronogen. We examined ultrastructural finding with electron microscopy. Twenty male Wistar Albino rats were divided into four groups:GroupI Control(C):5 rats received of 0.1ml/100g normal saline(NS) intravenously(i.v) injection for 4 weeks;GroupII Glomerulonephritis(GN):5 rats received i.v injection of anti-Thy1.1(0.25 g/100g) at zero day. GroupIII Glomerulonephritis+FK506 (GFK):5 rats received i.v injection of anti-Thy1.1(0.25 g/100g) and then FK506(1mg/kg) for two weeks.GroupIV Control+FK506 (CFK):5 rats received i.v.injection of NS (0.1mg/100mg) and then FK506(1mg/kg) for two weeks.Serum creatinine,creatinine clearance and proteinuria were performed at the end of the study period.Renal tissue were assessed for light microscopic findings glomerul and tubulointerstitial injury.PCNA, -SMA,VEGF,Laminin,MAC and

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apoptosis were semiquantatively scored on glomerul and tubulointerstitial area.We examined renal tissue by electron and immunofluorecence microscopy. Results At the end of the study period,glomerular cell proliferation and tubulointerstitial injury were significantly increased in GN compared to G group. Glomerular immunohistochemical expression of PCNA, SMA, VEGF, MAC,apoptosis significantly increased in GN compared to C group.Also we found the electron dense deposit on glomerular capillary walls and mesangium with IgG and C3 by immunofluorecence microscopy.The expression of the PCNA, -SMA and apoptosis were significantly decreased in GFK group compared to GN.But glomerular immunoexpression of MAC and VEGF were similar in GN and GFK groups.Also we found less proteinuria in GFK group compare to GN. Conclusion The present study suggests that FK506 may be useful in drug for the treatment of mesangial proliferative glomerulonephritis. PP4-70 INVESTIGATION OF PROTECTIVE EFFECT OF PREDNISOLONE IN NEPHROPATHY INDUCED BY CO-TRIMOXAZOLE IN RAT Ali Reza Mozaffari1, Iran Rashidi2 1 Internist, Department of Pathology-Ahwaz- Jondishapour Medical University, Iran 2 Pathologist, Department of Pathology-Ahwaz- Jondishapour Medical University, Iran Co-trimoxazole is a sulfonamide derivative, which is used as an antibacterial drug. Prednisolone is a dermo-corticostroid derivative, which is widely used as an anti-inflammatory and in the treatment of allergic reaction and collagen vascular diseases. Methods and Materials: The aim of this study was to find out the effect of co-trimoxazole on the renal interstitum and protective effect of predenisolone in rat. Four groups of animal were selected, namely A, B, C and control group. Groups A and B were treated with co-trimoxazole (150mg/Kg sulfa methoxazole + 30mg/Kg trimethoprim) while group C was treated with Co-trimoxazole and prednisolone (4mg/Kg) for 10 days. The reference group was only received some dose of water. The blood samples were collected from groups A and C 24 hours and from group B 14 days after the last dose of drug were administrated. Microscopic samples from kidney tissue were prepared for histopathological study. Results: The creatinine level in serum of group A (1.54) showed a significant increase as compared with the control group (1.04). In the group that received co-trimoxazole the histopathological study showed acute interstitial nephritis (AIN). In the group that treated with co-trimoxazole + prednisolone were not seen any histopathological changes . Conclusion: These results support the theory that AIN is drugs allergic reaction when co-trimoxazole is used. PP4-71 COX 2 AND CD 34 EXPRESSION IN WILMS TUMOR (NEPHROBLASTOMA) Gokben Yıldırım Kupesiz1, Bahar Akkaya1, Gulsun Tezcan2, Alphan Kupesiz2, Betul Unal1, Hakan Gulkesen3, Volkan Hazar2, Gulten Karpuzoglu1 1 Akdeniz University, School of Medicine, Department of Pathology, Antalya, Turkey 2 Akdeniz University, School of Medicine, Department of Pediatrics, Division of Hematology-Oncology, Antalya, Turkey 3 Akdeniz University, School of Medicine, Department of Statistics, Antalya, Turkey BACKGROUND Wilms tumor (nephroblastoma) is the most common primary malignant renal tumor of childhood. It occurs in approximately 10 children per million under the age of 15 years and is usually diagnosed between the ages of 2 and 5 years. Approximately 90–95% of Wilms tumors involve one kidney. There has been much recent interest in the role of cyclooxygenase-2 (COX-2), one of the three enzyme isoforms

that convert arachidonic acid to prostaglandins, in tumor development and progression. COX-2 expression is nearly ubiquitous in human cancers and has been correlated with poor prognosis [e.g., in tumors of the breast, cervix, colon, brain, and ovary]. Intriguingly, COX-2 seems to have multiple functions in tumor pathogenesis and thus represents an attractive therapeutic target. Among these, COX-2 seems to play a significant role as a positive regulator of tumor angiogenesis. METHOD Wilms tumor cases included to this study were obtained from archives of Akdeniz University Department of Pathology between 1997 and 2007. Immunohistochemical expression of COX -2 and CD 34 were independently evaluated and scored by two surgical pathologists. 17 patients (9 male / 7 female) were included in our group and their ages ranged from 1- 6 (Mean age 3.2). RESULTS In our study there was a significant negative correlation between angiogenesis, as measured by CD 34 expression and COX-2 expression (p=0.018, r=-0.567). There was no significant statistical correlation between gender, age and immunohistochemical results (COX -2 and CD 34 expression). There was also no difference between genders according to age. CONCLUSION In our study there was a significant negative correlation between angiogenesis, as measured by CD34 expression, and COX-2 expression. This would contradict the hypothesis that COX-2 acts as a pro-angiogenic stimulant, at least in Wilms tumor, and could suggest that COX-2 inhibits new blood vessel formation. Angiogenesis is regulated by a complex series of molecular pathways which, whilst they include the modulation of VEGF via PGE2 produced by COX-2, are subject to many other modulatory factors. Our study raises the possibility that COX-2 may influence tumor progression in Wilms tumor through mechanisms other than the promotion of angiogenesis. PP4-72 SEVERE RENAL VASCULITIS IN A PATIENT WITH DRESS Mehrenberger Marion1, Guitard Joëlle1, Ribes David1, Kamar Nassim1, Esposito Laure1, Rostaing Lionel1, Delisle Marie-Bernadette2, Modesto-Segonds Anne2 1 Department of Nephrology, Dialysis and Transplantation, CHU Rangueil, Toulouse, France 2 Department of Pathology, CHU Rangueil, Toulouse, France Introduction: Drug Rash with Eosinophilia and Systemic Symptoms (DRESS), is a severe generalized hypersensitivity reaction, characterised by fever, rash and internal organ involvement, such as liver and less frequently kidneys. It may be lethal in 10% of cases. Herein, we report on a case of a patient, who developed DRESS with renal insufficiency and proteinuria after allopurinol therapy, with the presence of a severe unexpected vasculitis at renal biopsy. Case Report : A 55 years-old woman, with a known impaired renal function due to nephroangiosclerosis, was admitted in 02/2006 for a gout crisis. Her serum uric acid, and creatinine levels were respectively 149 mg/l and 180 μmol/l. Allopurinol was introduced, but rapidly stopped because of myalgia, headache, and gastro-intestinal disorders. At that time, serum creatinine level was 188 μmol/l, with no proteinuria. Because of persistant hyperuricemia, allopurinol was re-introduced one month later. Eight weeks later, she presented with a diffuse maculopapular rash, fever (39°C), myalgia, and mild hypertension. Laboratory studies yielded the following results: serum creatinin level at 389 μmol/l, proteinuria at 0.8 g/d without hematuria, eosinophil count was 1192 / mm3, with mild cholestasis and C-reactive protein was 291 mg/l. The kidney biopsy revealed the presence of a severe leucocytoclasic necrotizing vasculitis with gigantocellular granulomas. There was no glomerulitis and no immune deposits. The diagnosis of DRESS with immuno-allergical vasculitis related to allopurinol was evocated. All immunological, bacterial and virological causes were ruled out. Allopurinol was stopped, and steroids initiated at the daily dose of 1 mg/k. Seven days later, serum creatinin level decreased to 220 μmol/l and cutaneous lesions

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disappeared, but proteinuria remained elevated (1.6 g/d). Two months later, the patient presented with a nephrotic syndrom and severe hypertension. A second renal biopsy revealed interstitial fibrosis and tubular atrophy, with regression of vasculitis lesions. Unfortunately, 4 months later, hemodialysis had to be started. Conclusion: DRESS must be promptly recognized and all potential culprit drugs whithdrawn, such as aromatic anticonvulsants, sulfonamides, calcium channel blockers, ranitidine, thalidomide and allopurinol. A kidney biopsy should be performed in patients suspected to have DRESS with impaired renal function and/or proteinuria. It may reveal the presence of immuno-allergic vasculitis that may evolve rapidly to end stage renal disease despite steroid therapy. PP4-73 RENAL AMYLOIDOSIS–CORRELATION MORPHOLOGY AND CLINICAL FEATURES Fulya Cakalagaoglu1, Aygun Ikinci1, Mehmet Koc2, Hakki Arikan2 1 Marmara University Pathology Department, Istanbul, Turkey 2 Marmara University Nefrology Department, Istanbul, Turkey BACKGROUND Amyloidosis was defined as the deposition of proteins that have the capacity to form beta-pleated sheets.The most common presentation of AA amyloidosis is that of renal diease.AL amyloid also frequently involved the kidney.In these study,we investigated whether different patterns of amyloid deposition occur in the kidney and these patterns can be related to the clinical findings. METHOD:29 cases of systemic amyloidosis with renal biopsies processed and examined histopathologically at the Department of Pathology, Faculty of Medicine University of Marmara were reviewed.Also all cases were stained with Amyloid A component(Ab-1;MS-1219-R7;Neomarkers;Fremont CA;USA)and Amyloid P protein(Ab-1;Neomarkers;RB-1786-R7 Fremont CA,USA)using the Standard avidin–biotin–complex immunoperoxidase method.The chemical types of amyloidosis were categorized according to the following staining reactions:AA amyloid was permanganate–sensitive and immunoreactive for AA protein.AL amyloid was permanganate resistant and not immunoreactive for AA protein.Renal amyloidosis were categorized glomerular,vascular and tubuloınterstitial by semiquantitive.We analyzed renal histologic findings,type of renal amyloidosis and clinical data. RESULTS.The age range of patients was 25-75 years.The male-female ratio was 19:11.3cases were categorized as AL and 26 were AA amyl.There was a predominence of the glomerular pattern in all cases.Also glomerular amyloid deposition was divided on the basis of morphological characteristics,into 4 types:a mesangial nodular type showing nodular mesangial deposits with sparse capillary wall involvement(16cases),a mesangio-capillary type disclosing diffuse amyloid in the mesangium(2cases), perimembranous type(6cases)and a hilar type showing amyloid deposits almost exclusively in hilar arterioles(3cases).14 cases of mesangial nodular type showed amyloid protein of AA type.Also we were compared glomerular amyloid deposition type and fibrosis.We found statistically significant correlation between hiler type deposition and fibrosis.We were showed that if patients had glomerular and tubulointerstitial deposition fibrosis would be worse.Proteinuria was the main clinical manifestation,present in all cases.Chronic renal failure and renal death appeared more common in hilar type of glomerular deposition with that of tubulointerstitial amyloid deposition. CONCLUSIONS.The results obtained suggest that the chemical type of glomeular amyloid protein is associated with significant differences in the morphological and prognostic features of the renal involvement.

PP4-74 MORPHOLOGICAL CHANGES AFTER LONG USAGE OF HEROIN THROUGH INJECTION Alexander Alexandrov1, Georgi Gergov1, Elka Ivanova1, Stanislav Hristov1, Todor Todorov2, Adrian Palov3, Dimka Hinova-Palova3, Tania Alexandrova4 1 Center of Forensic Medicine and Deontology - “Alaxandrovska” UMBAL, Sofia, Bulgaria 2 Center of Clinical Pathology - “Alaxandrovska” UMBAL, Sofia, Bulgaria 3 Department of Anatomy, Histology and Embryology at Medical Faculty, Sofia, Bulgaria 4 Emergency Medical Center – Vidin, Bulgaria

For the period 2003-2006 in the Centers of Forensic Medicine and Deontology and Clinical pathology at “Alaxandrovska” MBAL - Ltd., Sofia we had examined 33 patients (21 male and 11 female) that had died after a longtime intravenous usage of heroin and 23 patients with heroin dependency and clinical evidence of kidney damage who had undergone puncture biopsy (duration of the heroin dependency – 6 to 96 months). In order to determine the type of damages in the organism done directly by the toxic substance we studied macro- and microscopical (including electronic microscopical) morphological changes in different organs and systems – skin, underskin and underlying blood vessels – in the areas of the needle punctures (recent and old); lungs; heart; liver; kidneys and spleen. In all of the examined cases we discovered focused inflammatory changes on the skin and the underlying soft tissues in the areas most often used for the injection application of the narcotic. The most often morphological change seen was the fatty dystrophy. More rarely we encountered hepatitis with toxic origin, without the presence of fibrose changes in the liver parenchyma. The histological changed of these toxic liver damages are presented by vacuolization in the cytoplasm of the hepatocytes (fatty dystrophy), inflammatory infiltration of mononuclear cells and segment-nuclear leucocytes in the portal spaces of the liver (toxic hepatitis). In some cases there were inflammatory changes in the lungs, most often nonspecific (bronchitis, brohiolitis and focused pneumonia). In a particular case the morphological examination proved the presence of pneumonia caused by Pneumocystis carinii, damage of the lungs described in specialized literature by many leading authors. We succeeded in proving the agent of pneumocystis pneumonia not only by histochemical examinations (silver impregnation by Gomori-Grocott and PAS-reaction) but also at ultrastructural level by means of electronic microscopical examination. Kidneys are often affected in cases of longtime heroin usage. The percentage of the sclerotic glomerules of drug addicts, including those examined by us, is much higher than the one of people not addicted to drugs. The damage of the tubulo-interstitial apparatus (fibrosis, inflammatory interstitial infiltration and atrophic channels) is also a frequent morphological discovery in such individuals. Rarer are the kidney changes of the type of the glomerulopathies.

PP4-75 PHENOTYPIC ABERRATIONS IN MUCINOUS TUBULAR AND SPINDLE CELL CARCINOMA Alexandrou Paraskevi, Liapis George, Evangelou Kostas, Kyrıakou Vasiliki Pathology Department, Laiko General Hospital University of Athens, Greece

Mucinous tubular and spindle cell carcinoma (MTSCC) is a rare recently described variant of renal cell carcinoma with unique histological patterns which are sufficient to be designated by the World Health Organization as a distinct entity. We present a case of 55 year-old male who underwent a radical nephrectomy due to a tumor confined within the lower pole of the kidney. In gross examination the tumor was fairly circumscribed, measured 8.5 cm in greatest diameter and was tan-white in appearance. Microscopically the carcinoma was composed of variable proportions of tubular, tubulocystic, cord-like structures and

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spindle cells together with rare compact areas in the presence of extracellular blue gray mucinous background (Alcian blue +). The tumor cells were cuboidal to columnar with focally atypical nuclear features with either no or rare mitoses. No tumor necrosis was found. Apart from the predominant components, unusual growth patterns and features were focally identified such as pseudo-papillae, well formed papillae with psammoma calcifications, multinuclear neoplastic cells, glomeruloid structures, hemangiopericytoma, oncocytoma and clear cell carcinoma – like areas. The tumor cells showed a diffuse cytoplasmic staining for vimentin, Cam5.2, CK7, CK18, CK19 and racemase. Both tubular and spindle cell areas were negative to Keratin 34BE12, CD10, CD15, and chromogranin. The diagnosis of MTSCC was based principally on the predominant morphologic features along with the immunohistochemical profile, consistent with the 2004 WHO classification pathologic criteria. Since there is a relatively small number of published cases of MTSCC, neither the morphologic variabilities nor the histogenesis have been elucidated. The current case highlights the aberrant phenotype of the tumor which may contribute to the establishment of the diagnostic pathologic criteria.

PP4-76 PRIMARY SYNOVIAL SARCOMA OF THE KIDNEY. PRESENTATION OF TWO CASES IN CHILDREN Eugenia Garcia-Fernandez, Elena Ruiz-Bravo, Marta Mendiola, Purificacion Garcia-Miguel, Maria Luz Picazo La Paz University Hospital, Spain

Background: Renal primary sarcomas represent 5 % of kidney infantile tumors. Clear cell sarcoma (CCSK) is the most common in children. In 1995 Beckwith described a series of primary embryonal renal sarcomas with mesenchymal monophasic embryonal cells. This tumor was morphologically consistent with monophasic spindle synovial sarcoma (SS) and harbored the same SYT-SSX gene fusion. These tumors are very rare in children with only 5 cases reported in the literature. METHODS: Two cases of renal pediatric SS were identified in the files of the Department of Pathology in our Hospital. Histological, immunohistochemical and molecular analysis were performed in both patients. RESULTS: Patient nº 1: An 8-year-old boy had a history of abdominal pain and hematuria. Abdominal CT revealed a right renal mass with necrosis and vena cava thrombosis. Under the impression of nephroblastoma he started systemic chemotherapy and afterwards a radical nephrectomy with vena cava thrombectomy. The specimen weighted 292 grs and had a multicystic tumor that measured 8×7cm. Histologically the tumor showed a mesenchymal proliferation of short monomorphous cells. This proliferation entrapped tubules; some were cystically dilated and had cells with a hobnail morphology. After the surgery the patient had intensive systemic chemotherapy and radiotherapy. He remains free of disease 6 years after. Patient nº 2: A 2-year-old girl with a mass that arose from the inferior side of the right kidney. Systemic chemotherapy and nephrectomy were performed. The mass weighted 568 grs and measured 11×8,5cm, was well circumscribed, white-tan with small cysts and had less than 20% of necrosis. Histologically, the tumor showed a hypercellular mesenchymal proliferation. The nuclei of the cells were round with an average mitotic rate of 20/10hpf. Immunohistochemical studies showed diffuse positivity with vimentin, CD99 and bcl-2. Both cases were confirmed by molecular analysis of the characteristical t(X; 18) that results in the fusion of SYT gene to the SSX1 or SSX2 genes. CONCLUSSION: The differential diagnosis of SS in children includes CCSK, mesenchymal nephroblastoma, congenital mesoblastic nephroma and cystic nephroma. The differential histological diagnosis in the case of CCSK is sometimes difficult. The positivity of CD99 and bcl-2 is helpful in distinguishing SS from other tumors. However confirmation of this entity is done by molecular analysis. These renal tumors have been described in young adults, but exceptionally in pediatric patients, with only 5 cases described in the literature.

PP4-77 COLOR CLASSIFICATION AND AUTOMATIC REGION IDENTIFICATION IN GLOMERULAR PATHOLOGY Irina-Draga Caruntu, Simona Eliza Giusca Gr. T. Popa University of Medicine and Pharmacy, Romania Backgroud: The paper presents a new computerized technique for region identification in microscopic fields. It is based on the automatic separation of several chromaticity classes that are associated with different types of entities. Method: The method has been implemented by software modules developed in the Matlab environment and operates with indexed images. The material consisted of specimens from renal corpuscle biopsies, stained for light microscopy with haematoxylin-eosin and light green trichrome (Merck, Germany). The resulting microscopic fields exhibit a wide variety of colors, which have to be exploited in order to recognize the renal corpuscle elements: nuclei, connective tissue corresponding to the mesangium and fibrous crescent, Bowman space and capillaries (possibly containing blood cells). Results: The key issue of the proposed method is the iterative construction of a color map, whose entries are grouped in chromaticity classes, corresponding to the types of elements to be identified. Initially, each class contains a small number of representative colors selected by the user. Then, the classes are expanded, each iteration adding at least one new color to one or several of the existing classes. This step-by-step augmentation of the color map is automatically guided by a reciprocal validation of the included color(s) in the sense of the minimal distance computed in both the red-blue-green (RGB) and luminance-hue-saturation (YIQ) spaces. A number of images are considered for illustrating the correctness of the identified regions. These regions can be further used for quantitative analysis. Conclusions: The applicability of the technique is not limited to the renal corpuscle elements, but our focus on renal corpuscles is motivated by the difficult classification of the colors typical to such microscopic fields. It is worth mentioning that before elaborating this technique, we unsuccessfully tried to adapt segmentation procedures reported in literature, including the thresholding approaches experienced by us in some previous works. All these procedures failed because of the complexity of analyzed color images. PP4-78 CLASSIFICATION AND SCORING SYSTEM OF RENAL AMYLOIDOSIS; EXPERIENCE WITH 288 CASES Sait Sen, Banu Sarsik, Ayse Yazici Ege University, Medical School, Department of Pathology, Izmir, Turkey Amyloidosis refers to the extracellular deposition of fibrils composed of a variety of proteins, is multisystemic disease, associated with a different conditions. 25 different precursor fibrils are known up to date, of which seven fibrils were important for kidney. Although a variety of deposition patterns of amyloid have been described there are no universal renal classification and scoring system. This is necessary for predicting patient outcome, organ failure and also to establish an objective interpretation for clinical trials. In the present study, renal amyloidosis classification and scoring system were developed and applied. Cases of renal biopsy-proven amyloidosis were retrospectively evaluated (between 1990 and 2007). Amyloid was diagnosed using the stringent, alkaline, alcoholic CR staining method of Puchtler et al. The characteristic green birefringent polarization was taken as proof of the presence of amyloid. Glomerular amyloid depositions were classified similar to SLE 2003 ISN/RPS classification (class I to VI). Renal amyloid depositions (glomerular pattern and percent, vascular and interstitial) interstitial fibrosis, inflammatory infiltration and glomerular sclerosis were also scored. Renal amyloid prognostic score (RAPS) was found calculation of these scores. RAPS was divided into three stage (early, moderate and advanced

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amyloidosis). Total 305 renal (294 native, 11 transplant) biopsies from 288 patients were reevaluated. AA was detected in 90% of the patients. Biopsies from nine cases are inadequate for classification and staging. Depositions were evaluated as 10%, 18%, 22%, 38%, 1%, and 8% respectively for class I to VI. RAPS stages were found as 14%, 51% and 31% respectively for stage I –III. Clinical validation of this system has not been completed. We believed that renal biopsy is most suitable tissue for assessing of amyloid deposition intensity. Our classification and scoring system was applied successfully. Clinical validation studies are required to confirm importance and reliability. PP4-79 IMMUNOHISTOCHEMICAL AND MORPHOMETRICAL ANALYSIS OF THE TUBULOINTERSTITAL CHANGES IN PRIMARY GLOMERULOPATHIES Slavica Kostadinova-Kunovska1, Gordana Petrusevska1, Vesna Janevska1, Rubens Jovanovic1, Ladislava Grcevska2, Momir Polenakovic2 1 Institute of Pathology, Faculty of Medicine, Skopje, Macedonia 2 Department of Nephrology, Clinical Center, Skopje, Macedonia Alterations in the tubulointerstitial renal compartment were found in almost all glomerular diseases. The aim of this study was to analyse the tubulointerstitial changes in primary glomerulopathies and correlate them with the clinical data. Methods: We made a study on 50 renal biopsies with primary glomerulopathies and 20 control cases. For morphometric analysis we made a software color extraction of the interstitial area on tissue sections stained with trichrom Masson and expressed the results as percentage of the total scanned area. We made immunohistochemical analysis with Cytokeratin, HLA-DR, Vimentin, SMA and E-cadherin. We also applied double staining method, coupling Cytokeratin with Vimentin and SMA. Results: We found fibrosis, mainly focal, occupying more than 9% of the cortical tubulointerstitial surface (mean 18,75%) in 98% of the biopsies. In these areas the tubules showed marked atrophy. The tubular epithelial cells showed diminished positivity for Cytokeratin and E-cadherin in the areas of fibrosis, and enhanced expression of HLA-DR, unlike the control cases. The epithelial cells in the atrophic tubules, especially in the foci with inflammatory infiltrate, showed positivity for Vimentin and SMA, which was not visible in the control tissue sections. This was better visualized on the double stainings where both epithelial and mesenchymal markers were present in same tubular section. The morphometric analysis showed that the percentage of tubules with cells positive for Vimentin and SMA did not exceed 10% and 5%, respectively, of the total scanned area. The analysis showed statistically significant (p<0.01) positive correlation between the extent of fibrosis and the serum creatinine concentration. The extent of fibrosis was also positively correlated with the same statistical significance to the percentage of tubules with cells positive for Vimentin, SMA and HLA-DR. These parameters for tubular injury also show strong correlation with the serum creatinine concentration. Only the percentage of tubules containg cells positive for SMA was found to be correlated to the degree of proteinuria at the time of the biopsy. Conclusion: The tubular epithelial cells undergo epithelial-mesenchymal transition during various glomerulopathies, thus contributing to the interstitial fibrosis. The changes in the tubulointerstitial compartment influence the course of the disease. The quantitative histological analyses should be included in the process of nephropathological diagnosis in order to evaluate the histological risk factors in glomerular diseases.

PP4-80 AN ELECTRON-MICROSCOPIC (EM) AND LIGHT-MICROSCOPIC (LM) MORPHOMETRIC STUDY IN CHILDREN WITH GLOMERULAR IMMATURITY AND NEPHROTIC SYNDROME (NS) Aldona Wozniak, Malgorzata Janicka-Jedynska, Elzbieta Kaczmarek, Joanna Bulak-Joniec, Wieslawa Salwa-Zurawska Department of Clinical Pathomorphology, Karol Marcinkowski University of Medical Sciences, Ponza , Poland

Introduction: Very little information is available in the literature about the glomeruli maturation. It is thought that above 2 years of age the number of immature glomeruli should not exceed 10%. In children with NS glomerulopathies we encountered a small number of cases in whom immature glomeruli outnumber the percentage allowed to the given age. The aim of the study was: to compare the results of LM and EM evaluation and to carry out a morphometry. MATERIAL AND METHODS: The study group consisted of 66 children with NS. The diagnosis was established in LM and EM. The control group consisted of 30 children in the same age (with the same glomerulopathies MCD, DMH), but without glomerular immaturity. The morphometric study was performed. RESULTS: EM confirmed the diagnosis of MCD in 32, DMH in 6 and FSGS in 2 cases. The EM study did not confirm the LM diagnosis of MCD in 6 cases (5 DMH, 1 membranous glomerulopathy) and DMH in 1 case (MCD). EM revealed features of immaturity. There were differences in the intensity of foot processes effacement and microvillous transformation (between MCD and DMH). Morphometric studies revealed significant differences regarding the area involved by capillary loops (CLA) and glomerular volume. These values were significantly lower in the group with immature glomeruli, when the number of cells per 1000 m2 of CLA was higher. There were no correlations between the size of glomeruli and the age of children with glomerular immaturity (such correlations were evident in the control groups). PP4-81 PROGNOSTIC FACTORS IN DIFFERENT MORPHOLOGICAL VARIANTS OF RENAL CELL CARCINOMA (RCC) (ESPECIALLY IN RCC WITH SARCOMATOID FEATURES) Malgorzata Janicka-Jedynska, Aldona Wozniak, Jakub Zurawski, Elzbieta Kaczmarek, Joanna Bulak-Joniec Department of Clinical Pathomorphology, Karol Marcinkowski University of Medical Sciences, Ponza , Poland

Introduction: There is increasing incidence of RCC, when the biological behaviour may be different, thus commonly used prognostic factors are not always sufficient in predicting prognosis. Because of many possibilities of treatment, the choice of the best therapy is of importance and searching for better prognostic markers is necessary. Abnormal expression of E-cadherin and MUC1 has been associated with penetration by neoplastic cells. Inhibitors of cycline kinase (p21, p27) are responsible for antiproliferative properties. Morphological subtypes of RCC may contain sarcomatoid areas related with poor prognosis. The aim was to evaluate the expression of above mentioned markers with respect to histological and clinical features. MATERIAL AND METHODS: We investigated the expression of E-cadherin, MUC1, p21, p27 in 64 cases of RCC (5-7 years of follow-up): 32 conventional RCC (CRCC), 12 papillary (PRCC), 10 chromophobe (CHRCC) oraz 10 with sarcomatoid areas. RESULTS: The expression of E-cadherin and MUC1 substantially differed between studied groups. The strongest E-cadherin immunoreactivity was observed in CHRCC, less intense in PRCC and in CRCC. It was only focal in sarcomatoid areas. A diffuse cytoplasmic staining pattern for MUC1 was noted in CHRCC, whereas PRCC and CRCC showed predominantly membranous reaction. The appearance of strong cytoplasmic immunoreactivity for MUC1 in CRCC and

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sarcomatoid areas was related with higher grade of malignancy and poor prognosis. Differences with regard to p21 and p27 immunoreactivity were also observed. The lowest number of positively-stained nuclei was noted in CRCC and in sarcomatoid areas. Higher percentages were counted in PRCC and the highest in CHRCC. PP4-82 IMMUNOHISTOCHEMICAL STUDY OF TUBULOINTERSTITIAL INFLAMMATORY CELL INFILTRATION IN HUMAN GLOMERULONEPHRITIDES Gordana Petrusevska1, Slavica Kostadinova-Kunovska1, Ladislava Grcevska2, Vesna Janevska1, Rubens Jovanovic1, Momir Polenakovic2 1 Institute of Pathology, Faculty of Medicine, Skopje, Macedonia 2 Department of Nephrology, Clinical Center, Skopje, Macedonia The importance of the interstitial aspects of primary glomerular disease to overall renal function is continuously increasing. The aims of this study were to characterize the mononuclear inflammatory cells in the tubulointerstitium of the renal biopsies affected by glomerulonephritic lesions using a panel of monoclonal antibodies specific for leukocytic subpopulations. Correlations with tubulointerstitial changes and main clinical parameters of renal function were done. Methods: Renal biopsies from 50 patients with glomerulonephritides were studied. Ten control normal sections were obtained from kidneys removed for localized tumors. Paraffin sections were stained with HE, PAS, trichrom Masson, silvermethenamine Jones, as well as immunohistochemically with the following monoclonal antibodies: LCA, CD43, CD20, CD68, HLA-DR, E-cadherin. The morphometric analyses were done on sequentially taken images avoiding glomeruli and large vessels with an image analyzing system LUCIA M–NIKON. The extent of interstitial fibrosis was examined on trichrom staining. Results: 98% of the cases showed interstitial fibrosis higher than 9% of the surface of the total cross section, with a mean value of 18,75%. Mononuclear inflammatory infiltrate in the interstitium, mainly with focal distribution, was noticed in 90% of the cases. T lymphocytes were predominant (mean value 58,26%), the B cells were 18,62% and the macrophages were present with mean value of 22,92%. Positivity for HLA-DR showed 40,86% of the total inflammatory cells, a significantly higher value than the control group (23,5%). The tubular epithelial cells also showed higher expression for HLA-DR than the control group. The tubular cells, especially in cases with abundant inflammatory infiltrate manifested decreased expression for E-cadherin in comparison to the control group. Each of the inflammatory cells subpopulations, as well as the total inflammatory infiltrate showed strong correlations with the interstitial fibrosis. The clinical data showed strong association of the serum creatinine concentration with the number of cells from each of the leukocytic populations, as well as the total inflammatory cell count (Spearman R ranged between 0.44 and 0.47) and no correlation with the proteinuria. Conclusion: We can conclude that the interstitial inflammatory substrate may influence tubulointerstitial changes in the primary glomerulonephritides, as well as to determine the long-term prognosis of the disease, which implies further investigations in this field.

PP4-83 PRIMARY NON-HODGKIN LYMPHOMA OF URINARY BLADDER WITH RENAL INVOLVEMENT NINE YEARS LATER AND ABSENCE OF SYSTEMIC LYMPHOMA. A CASE REPORT Jasmina Markovi -Lipkovski1, Tatjana Terzi 1, Sanja Radojevi 1, Vesna emeriki -Martinovi 2, Vitomir Govedarovi 1, Gordana Basta-Jovanovi 1, Radmila Stevanovi 1, Dragan Mitrovi 1, Biljana Stojimirovi 3 1 Institute of Pathology, School of Medicine University of Belgrade, Serbia 2 Institute of Hematology, Clinical Centre of Serbia, Belgrade, Serbia 3 Institute of Urology and Nephrology, Clinical Centre of Serbia, Belgrade, Serbia Background. Primary bladder non-Hodgkin lymphoma (PBNHL) is very rare, especially as extranodal B-small lymphocytic lymphoma (B-SLL). Also, late isolated renal manifestation of PBNHL is extremely unusual. Case report. A 56-year-old woman was presented with a solitary tumor of bladder wall, with history of dysuria and night sweating. She underwent ultrasound, computed tomography (CT) and laparatomy which showed a solitary round tumor situated at the left lateral bladder wall. A transvaginal needle biopsy of the tumor was performed and diagnosis of primary extranodal B-SLL was made in the absence of bone marrow, lymph node or blood involvement. She was treated for 6 months with chemotherapy (LOP protocol) until achievement of complete remission, confirmed by CT and cystoscopy. Nine years later, she developed nephrotic syndrome and impairment of renal function. The renal tissue, obtained by percutaneous biopsy, was also diffusely infiltrated by small lymphoma cells with widespread tubulointerstitial destruction. Biopsy specimen showed a few glomeruli with focal segmental mesangial sclerosis and capsular adhesion, associated with segmental glomerular depositions of IgM detected by immunofluorescent microscopy. Thus, typical lesions for focal segmental glomerulosclerosis were observed. Immunohistochemical analysis of lymphoma cells infiltrating renal parenchyma and bladder wall revealed the same immunophenotype of lymphoma cell: LCA+, CD79a+, CD20+, CD5+, CD23+/ , CD43+/ , bcl-2+, CD3 , CD45RO , bcl-6 , Cyclin D1 , with very low proliferative activity (less than 5% Ki-67+ cells). A diagnosis of B-SLL was confirmed. Conclusion. Here we present a case of PBNHL with late isolated renal involvement, manifested with nephrotic syndrome. Widespread destruction of tubulointerstitium led to decrease of renal function. Glomerular changes typical for focal segmental glomerulosclerosis caused nephrotic syndrome. Glomerular lesions could be due to glomerular overloading phenomena because of large destruction of renal parenchyma with lymphoma cells. PP4-84 DIABETIC NEPHROPATHY ASSOCIATED WITH CRESCENTIC PROLIFERATION Eugen Mandache1, Mihaela Gherghiceanu1, Gabriel Mircescu2 1 ‘V. Babes’ National Institute of Pathology Bucharest, Romania 2 ‘C. Davila’ Hospital for Nephrology, Bucharest, Romania The purpose of this study was to investigate the cell types involved in the extra-capillary proliferation occurred in two cases of diabetic nephropathies. During the last couple of years, 14 renal biopsies with diabetic nephropathy have been thoroughly investigated for nephropathologic diagnosis. Two cases of already known diabetic patients have been found to develop extra-capillary proliferation featuring cellular crescents, segmental or circumferential, here and there developing epithelial, tubular profiles. Since the literature does not mention such an association, the problem raised is if this extra-capillary proliferation belongs to the diabetic pathology, or it is a

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consequence of some superimposed lesions. In one case the proliferation can be connected with some moderate IgA deposits, while in the second we could not find any other explanation. The crescentic proliferations have been thoroughly investigated in immunofluorescence, light and electron microscopy. One first remark was that all glomeruli with crescents had an intact basement membrane of Bowman capsule. The crescents contained only epithelial cells and fibrin in some places. This comes in agreement with the hypothesis that all monocytic cells penetrate the crescents from the periglomerular area through capsular breaks. A second remark concerns the glycogen content of proliferated parietal epithelial cells. While usually parietal epithelial cells are reach in glycogen rosettes, our samples showed crescents of clear cytoplasm cells with little glycogenic granules. The general conclusion was that in some conditions of diabetic nephropathy, injured glomeruli may develop extra-capillary proliferation with some distinct characteristics. PP4-85 COMPLEMENT C3 AND HYALINE VASCULAR DEPOSITS IN IgA NEPHROPATHY Mihaela Gherghiceanu1, Eugen Mandache1, Gener Ismail2, Mihai Voiculescu2, Sergiu Dumitrache3 1 ‘V. Babes’ National Institute of Pathology, Bucharest, Romania 2 Department of Internal Medicine-Nephrology, Fundeni Institute, Bucharest, Romania 3 ‘Carol Davila’ Hospital for Nephrology, Bucharest, Romania The aim of the study was to examine adult patients with IgA nephropathy (IgAN)and to analyze the effect of the clinical data and vascular lesions on renal failure progression. The clinical and pathologic materials of 25 IgAN patients were analyzed. The clinical data and histological features were recorded for each patient. The semiquantitative quantification was done for complement C3c and C3d vascular deposits evaluation. Clinical parameters were assessed at the known onset of the disease, at the time of renal biopsy, at the last outpatient check-up. Statistical analysis was done by segmentation, using the CHAID algorithm, included in the SPSS software. Hypertension was identified as the risk factor with the greatest impact upon risk of progression to renal failure among these patients (p<0.0001). Severe vascular lesions were significantly correlated (p<0.005) with a renal failure progression. Arteriolar C3c deposits were observed in 86.6% biopsies with IgAN, even in the biopsies from normotensive patients. Perinuclear vacuoles were observed in the smooth muscle cells in cases with C3c deposits in periglomerular arterioles in the biopsies from normotensive patients. The C3c and C3d deposition along the interstitial vascular wall was more severe in those with vascular lesions than in those without vascular lesions. C3d deposits were observed in arteries, arterioles and peritubular capillaries in all cases with hyaline vascular deposits. We have found focal arteriolar hyaline deposits, mostly in a nodular form, at the junction between arteriole and glomerular hilus in 48% of IgAN. The glomerular hilus was thickened and mesangium was enlarged at the vascular pole. Arteriolar hyaline deposits were correlated with development of the interstitial fibrosis and the percentage of C3d deposits area (morphometry – ImageJ). Vascular and tubulointerstitial lesions were identified as the most accurate histological prognostic factors regarding future evolution of IgA nephropathy. We suggest that complement activation at vascular level precede vascular lesions and subsequent hypertension. C3d vascular deposits and vascular hyaline reflect the disease degree and can be used as an important histological prognostic indicators.

PP4-86 SIGNIFICANCE OF CLINICAL PARAMETERS AND MORPHOLOGICAL ALTERATIONS IN ADULT-ONSET MINIMAL CHANGE DISEASE Kemal Kosemehmetoglu, Dilek Ertoy-Baydar Department of Pathology, Hacettepe University School of Medicine, Ankara, Turkey BACKGROUND: Minimal change disease (MCD) in children is the major cause of nephrotic syndrome with good prognosis, while adult-onset MCD has some differences such as slower response to corticosteroids and subsequent relapses. It has been reported that light microscopical and immunofluorescense may affect the response to steroid therapy and the long-term course. METHODS: Among the glomerular pathologies diagnosed by kidney biopsy between 1981 and 2006, we selected 18 adult patients whose biopsy result was the MCD. Clinical information was obtained from the hospital charts. The histologic sections were re-evaluated and morphological features were noted. RESULTS: The median age at diagnosis was 32.5 years, %66 of the patients were younger than 40. The male to female ratio was 1.25. Swelling of legs and periobital edema were the most common presenting symptom. Six patients had hypertension. The mean daily proteinuria was 3.95 g/day and serum albumin level was 2.5 mg/dl. Serum creatinine was high in 24% of cases. Urinalysis showed microscopic hematuria in 38% of patients, granular/hyaline casts in 57%. Hypertriglyceridemia and hypercholesterolemia were detected in 81% at presentation. In one patient, NSAID use was a suspect in etiology. The mean number of glomeruli per biopsy was 27 with 11% average ratio of global sclerosis. There were mesangial widening in 6 cases, trace amounts of immune deposits (C3, IgM or IgA) in 8. Arterial and/or arteriolar nephrosclerosis was noted in 9 patients. Five biopsies revealed mild interstitial inflammation. Clinical follow-up data were available on 12 cases. Mean follow-up period was 66 months (range, 8-204 months). Four patients experienced complete remission with no relapse. 5 patients had multiple relapses, 3 of which responded cyclosporine therapy. Time to relapse varied between 6 to 80 months. Presence of microscopic hematuria was the only parameter that is likely to correlate with the development of relapse (p=0.07). In all patients renal function was preserved after treatment. CONCLUSIONS: MCD is typically expected to show no abnormalities in light microscopy. However, there may be minor light microscopic alterations in some cases. In our study group composed of adult patients, neither the presence of these changes nor trace positivity of immune reactants predict the patient's clinical course or responsiveness to steroid therapy. Hematuria is the only clinical parameter which was likely to associate with the occurrence of relapse. Adult onset MCD has a good long term outcome with well-preserved renal function. PP4-87 HYDATIC CYST OF THE KIDNEY: A SERIES OF 39 CASES Fakhfekh Ines, Khabir Abdelmajid, Kallel Rim, Abbes Karima, Sellami Ahmed, Gouiaa Noures, Fakhafakh Hamadi, Boudawara Tahya EPS Habib Bourguiba SFAX, Tunisia Introduction: Kidney’s hydatic cyst was rare and characterised by its clinical characteristics and the therapeutic problems. The aim of this study was to identify the epidemilogic particularities, the diagnosis and therapeutic modalities of hydatic cyst in this exceptional localisation. Materials and methods We report a retrospective study relating to 39 cases of hydatic cyst of the kidney diagnosed and treated between 1982-2005 in Habib Bourguiba’s hospital, during the same period 2377 cases of hydatic cyst were operated in the same hospital. Results The male/femaleratio was 0,42. The ages ranged from 6 to 69 years (mean age 42,5). Clinically, the flank pain represented the most

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frequent symptomatology (86,5%), and cystic mass in 25 cases (62%), the hydaturia was present in 14% of cases. The intravenous urography highlighted a tumoral syndrome in 66,6% of the cases. Ultrasound evoked the diagnosis in 72,2% of the cases and classify the hydatic cyst according to Gharbi & coll’s classification in type I (18 cases), type II (5 cases), type III (8 cases), type IV (6 cases) type V (2 cases). Computed tomography was practised among 6 patients. The treatment was surgical in all the case and often conservative (85%), the nephrectomy was practised in 6 cases (15%). The diagnosis was confirmed by pathologic exam. Discussion The renal’s hydatic cyst accounts for 1.6% to 4% of all localizations in the human hydatidose. The clinical symptoms are variable. Imagery associated to serology allow to make the diagnosis. The treatment is surgical. PP4-88 VASCULAR ENDOTHELIAL GROWTH FACTOR IN RELATION TO NUCLEAR FACTOR – KAPPA B IN RENAL CELL CARCINOMA Gordana Dordevic1, Koviljka Matusan1, Emina Sinozic1, Vanja Licul2, Blazenka Grahovac1, Nives Jonjic1 1 Department of Pathology, School of Medicine, University of Rijeka, Croatia 2 Clinical Hospital Centre Rijeka, Croatia Objective: Renal cell carcinoma (RCC) is a malignancy with variable clinical course, partly attributable to specific genetic alterations of the different RCC types. Angiogenesis is important for tumor progression and metastatic spread. VEGF is a major factor which regulates angiogenesis while the nuclear factor-kappaB (NF-kB), a family of transcription factors; regulate a wide variety of cellular processes including cell growth, differentiation and apoptosis. The aim of this study was to evaluate the expression level of VEGF and to compare their values with the subcellular localization of NF-kB, proliferative rate of tumor cells and clinicopathological characteristics of RCC. Material and method: Immunohistochemical evaluation included VEGF expression, the subcellular localization of p65 member of NF-kB and Ki67 for proliferative rate of tumor cells in series of 16 RCC. Total RNA was extracted from the same tumor tissue samples, previously snap-frozen. Expression of VEGF was analyzed using quantitative Real-time PCR. Results: The difference in VEGF mRNA levels between conventional (11 clear cell RCC) and other histological types of RCC was not significant. Preliminary results indicate a more pronounce heterogeneity in VEGF expression within the group of clear cell RCC. VEGF mRNA levels show some positive association with VEGF protein expression evaluated as the percentage of positive cells and intensity of staining within the cells, while there was no association with NF-kB and Ki67. Larger tumors were characterized with higher value of VEGF expression, NF-kB and Ki67 and, moreover, the proliferation rate of tumor cells was higher in tumors with higher VEGF expression. Conclusion: Preliminary results indicate the heterogeneity in VEGF expression in conventional type of RCC. The association with tumor progression (tumor size and proliferation rate) indicates that VEGF is an important angiogenic factor in RCC. Although no association between NF-kB activity, VEGF expression and Ki67 was found on a small number of analyzed samples, our preliminary data suggest that the NF-kB may be important factor in renal carcinogensis by controlling cells proliferation. These findings need further validation of usefulness of NF-kB as a prognostic factor in RCC. PP4-89 COLLOIDAL IRON STAINING PATTERNS IN RENAL CARCINOMA TYPES AND CORRELATION WITH ELECTRON MICROSCOPIC FINDINGS Ipek Isik Gonul, Gonca Barit, Leyla Memis, Omer Uluoglu Gazi University School of Medicine, Department of Pathology, Ankara, Turkey

Background: Morphologically overlapping features of granular cytoplasm in clear cell carcinomas, chromophobe cell carcinomas and renal oncocytoma may cause difficulties in definitive diagnosis on light microscopic examination. Colloidal iron staining is considered to be characteristic for chromophobe renal cell carcinomas. In this study, our aim was to evaluate the colloidal iron staining patterns systematically in a spectrum of renal neoplasms and compare the findings with ultrastructural features. Method: We studied 116 cases of renal tumors which consisted of 93 clear cell carcinomas with different growth patterns and Fuhrman grades, 6 chromophobe cell carcinomas, 3 angiomyolipomas, 5 pelvic urothelial carcinomas, 5 papillary carcinomas, 2 oncocytomas and 2 collecting ductus carcinomas. One diagnostic paraffine section from each case was stained with colloidal iron by Hale’s method. Following determination of the staining patterns, the sites of sections with spesific staining were marked on the paraffine embedded tissue blocks and these areas were evaluated by electron microscopy after de-paraffinization and re-prosessing for electron microscopy. Results: We observed that other renal cell carcinomas apart from chromophobe cell carcinomas showed colloidal iron positivity. Four types of staining patterns were identified as follows: diffuse, strong, reticular staining, diffuse but weak, homogeneous cytoplasmic staining, focal, apical cytoplasmic, finely granular cytoplasmic staining and finally coarse granular cytoplasmic staining. Strong reticular staining was the only pattern observed in chromophobe cell carcinomas. On the other hand, other 3 patterns were seen in clear cell and papillary carcinomas together with renal oncocytomas. Urothelial carcinomas, angiomyolipomas and collecting ductus carcinomas were all negative. The ultrastructural features, particularly regarding the mitochondrial and microvesicular distribution were correlated with the light microscopic findings. Conclusion: Hale’s colloidal iron and the presence of numerous microvesicles on ultrastructural examination have been considered characteristic for chromophobe renal cell carcinomas. However, this study clearly demonstrated that clear cell renal carcinomas, oncocytomas and even papillary carcinomas could be diffusely positive for colloidal iron. Although staining pattern significantly differ in these tumors, light microscopic and histochemical findings sometimes require an ultrastructural confimation for definitive diagnosis. PP4-90 PATHOLOGICAL CHANGES IN KIDNEY BIOPSIES FROM PATIENTS WITH PLASMA CELL DYSCRASIA Stela Bulimbasic1, Arijana Racar-Pacic1, Kresimir Galesic2, Mirjana Sabljar-Matovinovic3, Ivana Kovacevic-Vojtusek4, Ivica Horvatic2, Danica Ljubanovic1 1 Department of Pathology, University Hospital Dubrava, Zagreb, Croatia 2 Department of Nephrology, University Hospital Dubrava, Zagreb, Croatia 3 Department of Nephrology, Clinical Hospital Merkur, Zagreb, Croatia 4Department of Nephrology, Vuk Vrhovac University Clinic for Diabetes, Endocrinology and Metabolic Diseases, Zagreb, Croatia Background: Renal impairment/insufficiency are common systemic complications of many hematological malignancies and they also occur in the majority of patients with plasma cell dyscrasia. To analyze incidence and clinico-morphological presentation of kidney involvement during plasma cell dyscrasia, retrospective analysis was performed. Methods: Patients with diagnosis of plasma cell dyscrasia with kidney involvement were identified from Renal non-tumor pathology Registry at Department of Pathology, University Hospital Dubrava. All the light microscopy glass slides (H&E, PAS, Masson, Jones and Congo special stain) were reviewed, along with material used for immunofluorescence and EM analysis. Results: During period

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from October 2003 to March 2007, 397 native kidney biopsies were analyzed. Kidney biopsies from 20 (5.04%) patients showed histological features consistent with diagnosis of plasma cell dyscrasia. This group included 7 male and 13 female patients. Patient age ranged from 38 to 76 years, median 60.9 years. All of them had proteinuria, which in 5 patients progressed to the nephrotic syndrome. An acute renal insufficiency were noted in 3 and chronic renal insufficiency in 4 patients. The most common histological patterns of renal involvement were AL amyloidosis (8 patients, 40 %) and cast nephropathy (7 patients, 35 %). In the remaining 5 patients, an acute tubular damage (3 patients, 15 %) and tubulointerstitial nephritis (2 patients, 10 %) were noted. Previous diagnosis of AL amyloidosis or immunoproliferative disorder/plasmocytoma had 4 and 3 patients respectively, while in the majority of patients, hematological evaluation and confirmation of diagnosis were completed after kidney biopsy. Discussion and conclusion: The plasma cell dyscrasia is frequently used as generic term for description of three distinct clinical entities: multiple myeloma, monoclonal gammopathy of unknown significance and dysproteinemia. They all have frequent renal involvement in common which can either present the first manifestation, or can occur later, during the course of disease. Morphological patterns of involvement vary, depending on which renal compartment is predominantly targeted. Commoner tubulopathic manifestation include Fanconi’s syndrome and cast nephropathy, while glomerular damage can be manifested as AL amyloidosis or monoclonal immunoglobulin deposition disease. Using combination of light microscopy, immunofluorescence and EM, all of them can be properly diagnosed in kidney biopsy, which is important for later clinical management and therapy. PP4-91 PRIMARY EXTRASKELETAL MESENCHYMAL CHONDROSARCOMA OF THE KIDNEY OF A YOUNG WOMAN: MORPHOLOGIC, IMMUNOPHENOTYPIC AND ULTRASTRUCTURAL ANALYSES: A CASE REPORT AND REVIEW OF THE LITERATURE Ozlem Yapıcıer1, Saime Sezgin Ramadan1, Abdurrahman Ozgur1, Kutlay Karaman2, Levent Turkeri3 1 Department of Pathology, Anadolu Health Care Center, Kocaeli, Turkey 2 Department of Radiology, Anadolu Health Care Center, Kocaeli, Turkey 3 Department of Urology, Anadolu Health Care Center, Kocaeli, Turkey Background: Extraskeletal mesenchymal chondrosarcoma is a rare tumor that has been reported mostly in neurosurgical and orthopedic literature. Five cases of primary renal extraskeletal mesenchymal chondrosarcoma are reported in the English literature with a variable, nonspecific presentation and relatively an aggressive behaviour. We present a case of primary extraskeletal mesenchymal chondrosarcoma of the kidney in a 28-year-old woman. We describe the radiologic, light microscopic, immunohistochemical and ultrastructural features of the case and discuss the differential diagnosis of the ekstraskeletal mesenchymal chondrosarcoma at this unusual site. Case: Following an episode of macroscopic hematuria, a 28-year-old woman underwent a radical nephrectomy for a radiologically confirmed right-sided renal mass. A radical nephrectomy was performed. Since the computed tomograms of the chest, abdomen, and pelvis showed no other lesions or evidence of metastatic disease, the histopathologic diagnosis is primary renal ekstraskeletal mesenchymal chondrosarcoma with its typical combined undifferentiated mesenchymal cells and cartilage islands. Urinary system CT revealed a relatively well-demarcated 3-cm tumor with calcifications and peripheral enhancement associated with foci of hypodensity within enhancing solid components. Microscopically, the tumor was composed of primitive undifferentiated mesenchymal cells and well-defined

islands of hyaline cartilage. The immunohistochemical study revealed diffuse vimentin positivity in the cytoplasm of the neoplastic cells. Pancytokeratin, epithelial membrane antigen, CD99, desmin and SMA were negative. S100 protein staining revealed dispersed positivity in cartilaginous areas. An electron microscopic study showed primitive precartilaginous mesenchyme displaying focal cartilaginous differentiation. Conclusion: The radiologic and the macroscopic aspect, together with the dual morphologic characteristics of differentiated cartilage islands interspersed within vascular undifferentiated mesenchyme, confirmed the diagnosis of primary extraskeletal mesenchymal chondrosarcoma of the kidney. In spite of its rarity, it is important to diagnose primary mesenchymal chondrosarcoma in kidney because its biological behaviour may be different from that of tumors of similar morphology. The differential diagnosis includes PNET/Ewing sarcoma and other rare sarcomas of kidney. The patient is free of disease nearly 13 months after its initial presentation. PP4-92 ULTRASTRUCTURE OF GLOMERULAR DEPOSITS IN CRYOGLOBULINEMIA Anastazija Hvala, Alenka Vizjak, Dusan Ferluga Institute of Pathology, Faculty of Medicine University of Ljubljana, Ljubljana, Slovenia Aims: Cryoglobulins are serum immunoglobulins precipitating at lower temperatures. They have been described as having a crystalline/fibrillar configuration. Our experience is not in line with such a description. Therefore, the aim of this study was to reevaluate systemically our kidney biopsy material with a particular emphasis to the ultrastructure of glomerular immune deposits in patients with primary and secundary cryoglobulinemia. Methods: In our archived bioptic material of 1992 kidney biopsies there were 18 biopsies of 14 patients with primary cryoglobulinemia (210-3600 mg/l) and 30 biopsies of 19 patients with autoimmune systemic connective tissue diseases, mostly systemic lupus erythematosus (SLE), and secondary mixed cryoglobulinemia (124-3300 mg/l). All biopsies were examind by light, immunofluorescence and electron microscopy (EM). For EM tissue samples were fixed in OsO4 and in some cases in formaldehyde, embedded in Epon 812 and stained with uranyl acetate and lead citrate. EM analysis of immune deposits was performed on highly magnified photos. Results: Deposits of various electron density were observed in glomeruli of all biopsies. In primary and in SLE associated cryoglobulinemia glomerular capillary wall deposits were found to be homogeneous or finely granular, with no evidence of any fibrillar structure. In 4 patients with SLE besides homogeneous, fingerprint deposits were also demonstrated. Glomerular and extraglomerular fingerprint deposits were demonstrated in two patients with low cryoglobulin levels, one with primary Sjögren’s disease and the other with non-classified autoimmune systemic disease. In a patient with glomerular monoclonal IgG kappa deposits showing an ultrastructure of randomly oriented bundles of parallel fibrils described as characteristic for cryoglobulinemia type I, no detectable cryoglobulins were found in the serum. Conclusion: Our detailed EM study on kidney biopsy specimens reveals that glomerular immune deposits in patients with primary and secondary mixed cryoglobulinemia can be of various electron density, but usually homogeneous or finely granular. Furthermore, our study suggests that cryoglobulin deposits display an organised substructure with crystalline/fibrillar configuration, described originally in 1977 by Feiner and Gallo and frequently cited as characteristic, only occasionally in a minority of patients with cryoglobulinemia.

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PP4-93 FSGS VARIANTS: CORRELATION OF HISTOPATHOLOGICAL FEATURES AND CLINICAL PRESENTATION Guldal Yilmaz, Saba Kiremitci, Arzu Ensari Ankara University Medical School, Department of Pathology, Ankara, Turkey Background: Focal segmental glomerulosclerosis (FSGS) is a clinicopathologic syndrome characterized by nephrotic syndrome and glomerular sclerotic lesions. Recently, several morphologic variants of primary FSGS which have different prognostic and therapeutic implications were defined. These variants are (1) FSGS-Not Otherwise Specified (NOS), (2) FSGS-Cellular variant (CeV), (3) FSGS-with diffuse mesangial hypercellularity (DMH), (4) FSGS-Collapsing variant (CoV), (5) Glomerular tip lesion (GTL), and (6) C1q nephropathy. We have, therefore, decided to categorize the FSGS cases diagnosed in our Department and evaluate their clinical significance. Method: Renal biopsy samples of 48 cases of primary FSGS, which are stained with Hematoxylin and Eosin, periodic-acid Schiff, methenamin silver and trichrom, were revised and categorized according to the morphologic variant grouping. All cases were also examined by IFM and EM. Clinical data was retrieved from the patient files. Besides FSGS, the CeV is characterised by endocapillary hypercellularity occluding lumens while DMH has generalisad mesangial hypercellularity. The CoV has the wrinkled and collapsed glomerular basement membrane. GTL is defined by the presence of segmental lesions in the periphery of the glomerular tuft and synechia formation between the glomerular tuft and Bowman’s capsule at the tubular pole. C1q nephropathy is defined by FSGS with variable mesangial hypercellularity and dominant paramesangial deposits of C1q. Results: Among these 48 cases, 11 (%22.9) were categorised as GTL, 5 (%10.4) as FSGS-CeV, 2 (%4.2) as FSGS-DMH, 2 (%4.2) as FSGS-CoV, 3 (%6.2) as mixed form of GTL and FSGS-DMH, 25 (%52.1) as FSGS-NOS. None of the cases was categorised as C1q nephropathy. IFM and EM did not show any difference between the variants of FSGS. Retrieving the clinical data, it was found that collapsing variant presented with proteinuria alone, and FSGS-DMH with both proteinuria and hematuria. FSGS-NOS, GTL and mixed form presented with either proteinuria alone or with hematuria. FSGS-CeV, usually presented with proteinuria alone, but in one patient it was associated by hematuria. In one patient the clinical data was lacking. Conclusion: As recent studies showed that identifying the morphologic variants of primary FSGS may provide further prognostic and therapeutic implications clinically, the renal pathologists should try to evaluate the morphologic variants of FSGS. IFM and EM do not seem to be useful in the differential diagnosis of FSGS variants.

Pulmonary Pathology PP4-94 THE EXPRESSION OF E-CADHERIN IN NON SMALL CELL LUNG CANCER Chrisoula Tsobanidou1, Doxakis Anestakis1, Ioannis Dimitriadis1, Jakob Anjel2, Nikolaos Barbetakis3, Frideriki Patakiouta1 1 Department of Pathology, “Theagenio” Cancer Institute, Thessaloniki, Greece 2 Pulmonology Department, “Theagenio” Cancer Institute, Thessaloniki, Greece 3 Thoracic Surgery Department, “Theagenio” Cancer Institute, Thessaloniki, Greece

Background: Several diagnostic and prognostic markers are being used in recent years, in order to investigate the pathogenesis of lung cancer. E-cadherin is a related new, very promising marker in cancer research, which is not widely investigated in lung cancer. E-cadherin is important for cell-cell adhesion. Reduction and/or loss of E-cadherin expression in non-small cell lung cancer correlates positively with malignant transformation of tumor and with the potential of these tumors for invasion and metastasis. The aim of the present study was to examine immunohistochemically the expression of E-cadherin in non-small cell lung cancer (NSCLC) and their relationship with clinicopathologic parameters such as histological type, grade, tumor size, sex ad age of the patients. Material and Methods: Adenocarcinomas, squamous cell carcinomas and large cell carcinomas fall within the spectrum of NSCLC. 42 patients (19 women and 23 men) with NSCLC treated surgically were examined retrospectively. The tumor specimens have been immunostained for E-cadherin. The tumors were squamous cell carcinomas (12 cases), adenocarcinomas (18 cases) and large cell carcinomas (14 cases). The age of the patients ranged from 44 to 78 (mean age 65 years). Immunohistochemistry was performed using an avidin-biotin complex method and monoclonal antibody against E-Cadherin. Membrane and cytoplasmic staining in >50% of tumors cells was considered as a positive expression of E-Cadherin. Results: Impaired E-cadherin expression (loss or cytoplasmic delocalization <50% of cells) was observed in 36 (85,7%) of 42 samples. Absent or reduced expression of E-cadherin was observed in 10(83,3%) squamous cell carcinomas , 13(72,2%) adenocarcinomas and 13(92,8%) large cell carcinomas. The Impaired expression of the E-cadherin was observed with a higher frequency in high- grade (squamous cell carcinomas and adenocarcinomas) than in low grade tumors (squamous cell carcinomas, adenocarcinomas and large cell carcinomas) 82% and 48% respectively, (p<0.0001). No relationship was found between E-cadherin impaired expression and tumor size or sex of the patient. Absent or reduced expression was seen in advanced age of the patients. Conclusion: Impaired E-cadherin expression was fount more frequently in high grade and more aggressive histological type tumors and as well as in advanced age of the patients. PP4-95 EXPRESSION OF CD44 AND MMP-2: POSSIBLE ASSOCIATION WITH HISTOPATHOLOGICAL FEATURES OF INTRATHORACIC SOLITARY FIBROUS TUMORS Funda Demirag1,Ebru Cakir1,Sibel Alpar2,Irfan Tastepe2,Sadi Kaya3 1 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Turkey 2 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Department of Chest Diseases, Turkey 3 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Department of Chest Surgery, Turkey

Background: Recent researches had showed that tumor cell adhesion molecular CD44 and matrix metalloproteinases (MMP-2) were expressed strongly in many tumors, and was associated closely with invasion and metastasis of the tumors. Although solitary fibrous tumors (SFT) have a good prognosis , a minority

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bahave malignant. The aim of this study was to analyze CD44 and MMP-2 expression to question histopathological parameters. Method: We reviewed 10 patients who had undergone surgical resections for benign or malign SFT . Clinical findings and follow–up informations were obtained for all patients. Haemotoxylen-Eosin stained sections were reexamined for evalution of histopathological parameters (degree of cellularity, keloidal type collagen deposition, perivascular hyalinization, hemangiopericytoma-like area, nuclear pleomorphism, number of mitoses, necrosis, cyst formation, myxoid areas). All cases were positive for CD34. Immunostaining of CD44 and MMP-2 were performed by using the streptavidin-biotin method with mouse monoclonal antibody. The intensity of CD44 and MMP-2 were evaluated by light microscopy. Results: 10 patients underwent surgical resection . 8 SFT were benign and 2 SFT were malign. There were 3 men (%30) and 7 women (%70) of a mean age of 54.5 years (range 30 to 71 years). One patient had a history of asbestosis exposure . Complete resection was performed in all cases. The tumor arising from pleura was excised with pleurectomy. In one embedded intraparenchimal tumor, pneumonectomy was performed. According to England criteria, 2 malign SFT were contained multiple masses. 3 cases originated in the right hemithorax and 7 in the left. Tumor size ranged from 4.5-27 cm. Coagulative necrosis was observed in 2 benign SFT and 2 malign SFT. All cases expressed strong CD44. However only 2 malign SFT expressed focal MMP-2. Conclusion: Although MMP-2 positivity was observed in 2 malign cases, CD44 positivity was not associated with malignant criteria in solitary fibrous tumors. PP4-96 MUC-1 EXPRESSION IN BENIGN PULMONARY LESIONS AND NORMAL LUNG TISSUES Funda Demirag1, Ebru Cakir2, Hulya Bayız3, Saliha Battal3 1 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Turkey 2 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Deparment of Pathology, Turkey 3 Ataturk Chest Diseases and Chest Surgery Education and Research Hospital, Department of Chest Diseases, Turkey

Background: The mucin MUC-1, encoded on chromosome 1q21, is a high molecular weight transmembrane glycoprotein and is expressed normally at the apical borders of epithelial cells in the breast , colon, and lung. Expression of MUC-1 has several effects in cells, including inactivation of integrin mediated cell to matrix adhesion and E-cadherin mediated cell to cell adhesion as well as protection from cytotoxic lymphocyte. Our aims were to study association between the expression of MUC-1 and biologic nature benign pulmonary lesions and normal lung tissues. Method: We selected 4 intrapulmonary bronchogenic cyst, 2 sclerosing haemangioma, 2 usual interstitial pneumonia , 1 adenoid cystic malphormation and 16 normal lung tissues. Normal tissues were obtained from histologically normal lung tissue adjacent to carcinoma. Among the neoplatic tissues 9 were adenocarcinomas, 5 squamous cell carcinomas, 1 adenosquamous carcinoma and 1 mucinous type bronchioloalveolar carcinoma. Two normal lung tissues contained atypical adenomatous hyperplasia. MUC-1 expression was assessed by immunohistochemistry with monoclonal antibody against MUC-1 mucin. Membranous staining for MUC-1 was divided into complete and incomplete staining. Results: Apical surface of upper respiratory epitelium in all cases were stained completely with MUC-1. The surface cells in sclerosing haemangioma and atypical adenomatous hyperplasia were strong complete positive for MUC-1. The positivity with MUC-1 in normal tissues around the adenocarcinomas was stronger than squamous cell carcinoma. Conclusion: Complete MUC-1 positivity showed type II pneumocyte lineage. This feature will facilitate differential diagnosis of based type II pneumocyte pulmonary lesions.

PP4-97 HISTOLOGICAL FINDINGS IN CARDIOGENIC ALVEOLAR-CAPILLARY BARRIER DAMAGE Mauro Canzian1, Felipe Zampieri2, Edwin Parra2, Ronaldo Kairalla3, Vera Capelozzi2 1 Division of Pathology, Heart Institute (InCor), School of Medicine, University of São Paulo, SP, Brazil 2 Department of Pathology, School of Medicine, University of São Paulo, SP, Brazil 3 Division of Respiratory Diseases, Heart Institute (InCor), School of Medicine, University of São Paulo, SP, Brazil

Background: Chronic heart failure increases resistance to gas transfer across the alveolar-capillary interface. Acute pressure or volume overload can injure the alveolar blood-gas barrier. These alterations are generally reversible due to the reparative properties of the alveolar surface. However, when alveolar-capillary membrane is chronically challenged, remodeling of pathophysiologic and clinical parameters may take place. The resulting pulmonary histological changes in surgical lung biopsy specimens have been barely exploited. Method: We examined medical records of all cardiac patients with chronic heart failure, presenting lung diffuse infiltrates, who underwent surgical lung biopsy, from January 1982 to December 2005. Patients with congenital cardiopathy, infectious manifestations or under immunosuppressive treatment were excluded. Twenty-four patients met our study criteria. Histological features including alveolar collapse, edema, protein deposition, hemossiderin-laden macrophages accumulation and hemorrhage, venous and lymphatic ectasia, vascular sclerosis, capillary congestion and fibroblast foci were studied. Results: Twenty patients were men and 4 women, with a median age of 54 years (range 16 to 79 years). The main chronic heart failure causes were myocardial ischemia, heart valve diseases and dilated hypertensive and Chagas cardiomyopathies. Based on alveolar-capillary barrier changes three main groups were created. Group I comprised acute cardiogenic pulmonary edema, ranging from minimal “cuff” around of bronchovascular axis to severe alveolar fluid accumulation. The second group comprised chronic capillary congestion presenting different degrees of septal thickening, capillary dilatation and hemossiderin-laden macrophages alveolar accumulation. Group III were histologically characterized by diffuse alveolar damage, in focal or segmental distribution. In 3 of 5 patients belonging to group I, acute cardiogenic pulmonary edema was the main change due to myocardial ischemia. Among the 11 patients belonging to group II, 4 had myocardial ischemia whereas 3 had heart valve disease. Diffuse alveolar damage, found in 7 patients, didn’t show any clinical picture predominance. Conclusion: Alveolar-capillary barrier injury due to chronic pressure or volume overload is manifested through different patterns of histological changes which recognition can influence prognosis and treatment. Patients with chronic heart failure should be evaluated to establish treatment protocols for acute pulmonary edema, chronic passive congestion and diffuse alveolar damage. PP4-98 INTENSITY OF LUMINAL ALVEOLITIS IN PULMONARY TUBERCULOSIS Zdravko Kosjerina Institute of Lung Diseases, Serbia

Material and methods: Material of the study included the transbronchial biopsy samples obtained from 30 tuberculous patients. The quantity of inflammatory and immunocompetent cells was estimated by stereometric method numerical density. The term "perigranulomatous alveoli" is used to denote the alveoli surrounding a granuloma at the distance of 100 micrones from its external edge. The remaining alveoli are refered to as "the alveoli far away from a granuloma". Results: Numerical density of all cells found in the lumen of the alveoli far away from a granuloma ranges between 18.327/mm3 and 96.120/mm3,

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the mean value being 43.042/mm3. Lymphocytes predominate -21.778/mm3 (50.6%), followed by macrophages 19.916/mm3 (46.3%). Numerical density of all cells present in the lumen of perigranulomatous alveoli is found to range from 14.639/mm3 to 197.628/mm3, the mean value being 74.662/mm3. Lymphocytes predominate 42.460/mm3 (56.9%), followed by macrophages 29.555/mm3 (39.6%). Discussion: The presence of inflammatory and immunocompetent cells on the surface and in the lumen of the alveoli accounts for luminal alveolitis. The presence of these cells in the lumen of the perigranulomatous alveoli accounts for perigranulomatous luminal alveolitis which has a significantly higher intensity than the luminal alveolitis in the alveoli far away from a granuloma. Thus the intensity of perigranulomatous luminal alveolitis is found to be 74.662 cells/mm3, while the intensity of the luminal alveolitis in the alveoli far away from a granuloma is 43.042 cells/mm3. Conclusion: Perigranulomatous luminal alveolitis in tuberculosis is 1.7 times as intense as alveolitis in the alveoli far away of a granuloma. PP4-99 COMPARISON OF OSTEOPONTIN, -CATENIN AND HNRNP B1 EXPRESSION IN LUNG CARCINOMAS Muhammet Emin Guldur1, Yasemin Kibar2, Hale Deniz3, Kemal Bakir2 1 Gaziantep Gynecologic and Obstetric Hospital, Gaziantep, Turkey 2 Gaziantep University,Medical Faculty, Department of Pathology, Gaziantep, Turkey 3 Gaziantep Pediatric Hospital, Gaziantep, Turkey

BACKGROUND: This study was performed to compare osteopontin, -catenin and hnRNP B1 immunoreactivities in small cell lung carcinomas (SCLC) and non-small cell lung carcinomas (NSCLC). Correlation of these three antibodies with grade and stage in NSCLC was also investigated. METHODS: Twenty-nine SCLC, 6 large cell carcinoma, 36 adenocarcinoma and 30 squamous cell carcinoma (SCC), totally 101 cases, were included in this study. Osteopontin, beta-catenin and hnRNP B1 expressions were immunohistochemically evaluated. In addition, these immunoreactivities were compared with grade and clinicopathologic stage in NSCLC. RESULTS: Osteopontin positivity was 6.9% in SCLC and 67% in NSCLC. When NSCLC types were individually considered, osteopontin positivity was 66.7% in large cell carcinoma, 80% in SCC and 55.6% in adenocarcinomas. -catenin positivity was observed in 49.3% of NSCLC and none of SCLC cases. These results were statistically significant (p<0.05). No statistically significant difference was found in RNP immunostaning between SCLC and NSCLC (p>0.05). Neither grade nor stage of NSCLC was correlated with osteopontin, -catenin or hnRNP B1 immunoreactivity. CONCLUSION: We observed that osteopontin and -catenin are useful in differentiating SCLC from NSCLC. In this study, grade and stage of NSCLC were not found to be correlated with any of three antibodies. We propose that osteopontin and -catenin may be useful in discriminating NSCLC with neuroendocrine differentiation and high-grade neuroendocrine tumors of the lung. PP4-100 GLUCOSE TRANSPORTER-1 EXPRESSION IN PULMONARY NEUROENDOCRINE CARCINOMA rem Hicran Ozbudak1, Fabio Tavora2, Negar Rassaei2,

Konstantin Shilo2, Wei-Sing Chu3, Junya Fukuoka4, Jin Jen5, William D Travis6, Teri J Franks2 1 Department of Pathology, University of Akdeniz, School of Medicine, Antalya, Turkey 2 Department of Pulmonary and Mediastinal Pathology, Armed Forces Institute of Pathology, Washington, DC, USA 3 Department of Scientific Laboratories, Armed Forces Institute of Pathology, Washington, DC, USA 4 Laboratory of Pathology, Toyama Medical & Pharmaceutical University, Toyama, Japan

5 Laboratory of Population Genetics, National Institutes of Health, Bethesda, MD, USA 6 Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY, USA Background: Malignant cells are known to have accelerated metabolism, high glucose requirement and increased glucose uptake. Transport of glucose across the cellular membrane is mediated by facilitative glucose transporter proteins. Glucose transporter-1 (GLUT-1) is one of 14 members of this family and is typically not detectable in normal tissue or benign lesions. Elevated levels of GLUT-1 expression and/or activation have been shown to be associated with malignancy. Since only limited studies address the issue of GLUT-1 expression in lung carcinoma, we sought to investigate its expression in pulmonary neuroendocrine carcinomas. Method: Tissue microarray based samples of 178 neuroendocrine carcinomas, including 48 typical carcinoids (TC), 31 atypical carcinoids (AC), 27 large cell neuroendocrine carcinomas (LCNEC), and 72 small cell carcinomas (SCLC) from different patients were studied immunohistochemically for GLUT-1 (Polyclonal, 1/300, Dako, Carpinteria, CA) expression. A composite score including distribution and intensity of labeling was compiled from each core. Correlation of GLUT-1 expression with clinical-pathological variables was performed utilizing SPSS 13.0 (SPSS Inc., Chicago, IL, USA). Results: Forty seven percent (75/161) of pulmonary neuroendocrine carcinomas were immunoreactive with GLUT-1. Distinct membranous staining was observed in neoplastic cells with concentration along the luminal border of tumor islands. Normal bronchial epithelium and alveolar pneumocytes were negative. GLUT-1 expression correlated with tumor differentiation/type, p<0.001: 6.5% (3/46) cases of TC, 20.7% (6/29) of AC, 73.9% (17/23) of LCNEC, and 77.8% (49/63) of SCLC were positive for GLUT-1. Five year survival of GLUT-1 negative neuroendocrine carcinomas was 61% versus 21% GLUT-1 positive cases (p<0.001); however, this correlation was not independent of tumor type/grade. There was no correlation between GLUT-1 expression and gender, tumor size or stage. Conclusion: GLUT-1 is expressed in approximately half of pulmonary neuroendocrine carcinomas and displays membranous staining with predilection for the luminal surface of tumor islands. GLUT-1 shows strong correlation with neuroendocrine carcinomas differentiation/grade, but not with other clinicopathologic variables. PP4-101 NUCLEAR ROUNDNESS IN NEUROENDOCRINE TUMORS OF THE LUNG Zaklina Mijovic1, Dragan Mihailovic1, Milos Kostov2 1 Institute of Pathology, University of Nis, Serbia 2 Department of Pathoanatomy, Military Hospital, Nis, Serbia INTRODUCTION: The most recent WHO classification of pulmonary neuroendocrine tumors recognizes four entities: typical carcinoid, atypical carcinoid, large cell neuroendocrine carcinoma and small cell carcinoma. AIM: The aim of this study was to estimate nuclear size and roundness in carcinoid tumors and small cell carcinomas of the lung. MATERIAL AND METHODS: At Institute of Pathology, University of Nis five cases of typical carcinoid tumor and ten cases of small cell carcinoma of the lung were analyzed on biopsy samples obtained by fiberoptic bronchoscopy. After formaline fixation and paraffin embedding, serial histologic sections were routinely stained with H&E. The nuclear size et roundness were estimated using image analyzer LUCIA M 3.51 ab (Nikon) at objective 40x, after binary image editing. In each case a hundred nuclei were measured. A statistical analysis was performed using Mann-Whitney test. RESULTS: The roundness of nuclei in typical carcinoid tumor (0.963±0.007) was significantly larger than in small cell carcinoma of the lung (0.908±0.021), p<0.01. No significant differences in nucler size were found. CONCLUSIONS: The

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authors conclude that nuclear shape is more rounded in neuroendocrine lung tumors of low-grade -typical carcinoids in comparison to high-grade tumors -small cell carcinomas. Further studies on a larger number of patients are required to confirm these findings. Key words: roundness, carcinoid, small cell carcinoma. PP4-102 COEXISTING PULMONARY TUBERCULOSIS AND MULTIFOCAL PULMONARY CARCINOID IN THE SAME LOBE: A CASE REPORT Fillinger Janos1, Heiler Zoltan2 1 Koranyi National Institute for Lung Diseases Budapest, Hungary, Department of Pathology, Hungary 2 Semmelweis Medical University Budapest, Hungary, Department of Thoracic Surgery, Hungary The synchronous occurance of lung carcinoma and tuberculosis is well known , but coexistens with carcinoid tumor is very rare. One case was mentioned in the literature. In our Institute between 2003-2006 among 96 cases was one patient. A 38 years old woman has observed on screening chest X-ray multiplex nodules in lower left lobes. TTB was taken but cytological examination reveal necrosis.Bronchoscopy was negativ too.The patient underwent surgery for lobectomy. Intraoperativ cytology shows necrosis. Mediastinal lymph node dissection was performed simultanously. The histology showed tuberculosis acino-nodosa caseosa and multifocal appearence of typical carcinoid.With Zeel-Nielsen stain can be found some acid stain rod shape bacilli in granulomatous nodules. Immunhistochemical reaction in tumor cells was positiv with NSE,Chromogranin-A, Synapthophysin and panCytokeratin. Gatrointestinal examination looking for primary site for tumor was taken with negativ findings, serum chromogranin was in normal range. The patient has got antituberculotic therapy.Apropos of this case authors would like to present the diagnostic difficulties of multiplex nodules of lung. Which nodules must be investigated? PP4-103 EXPRESSION OF p63, K903, TTF-1 AND CK-7 IN LUNG CARCINOMAS AND DIFFERENTIAL DIAGNOSIS Zekiye Aydogdu Dinc1, Nur Yucel1, A. Kadri Cirak2, Gultekin Tibet2 1 Izmir SS Chest Disease and Surgery Hospital, Department of Pathology, Izmir, Turkey 2 Izmir SS Chest Disease and Surgery Hospital, Clinic of Chest Disease, Izmir, Turkey Background: Lung carcinomas are the leading cause of death by cancer in the world both males and females.Accurate morphologic dictinction between small cell (SCLC) and non-small cell lung cancer (NSCLC) has therapeutic significance. However evaluating morphology can be limited by crush artifact, tumor necrosis, limited tumor representation and overlapping morphologic features in small biopsies. Method:In this study, we evaluated a panel of antibodies , containing p63, K903, TTF-1 and CK7, for their efficiancy in distinguishing between morphologic types of lung carcinoma by immunohistochemical method. Formalin fixed parafin embedded tissue section of 23 squamous cell carcinoma (SCC),12 adenocarcinoma (AC), 5 large cell carcinoma(LCC) and 6 small cell lung carcinoma(SCLC), that belong 46 patients underwent surgery and bronchoscobic biopsy ( for only SCLC cases ) were used. Results:High proportion of SCC were positive with p63(% 94) and K903 (%88), whole SCLC were positive with TTF-1 (100) and most of AK were positive with CK7 (%76). Staining patern in tumor cells especially for p63 changed with tumor differantiation . Conclusion: Accurate histologic typing of lung tumors can be evaluate with a panel of immunohistochemistry, especially in the little biopsies that have limited morphologic features.

PP4-104 IMMUNOHISTOCHEMICAL ANALYSIS OF COX-2 EXPRESSION PATTERN IN SPORADIC LUNG CARCINOMAS OF NEUROENDOCRINE ORIGIN Christos Valavanis, Maria Lekarakou, Helen Siatra, Prokopis Vogiatzis, Maria Britsou, Joanna Lekka, Petroula Arapantoni-Dadioti Molecular Pathology Unit, Dept of Pathology, METAXA Cancer Hospital, Piraeus, Greece BACKGROUND-OBJECTIVE: Cyclooxygenases are proteins that mediate the conversion of arachidonic acid to prostaglandins and other biolactive lipids.Prostaglandins are involved in cell growth regulation, angiogenesis and metastatic process. Two COX isoforms have been identified COX-1 and COX-2. In normal cells the COX-1 is expressed constitutively whereas COX-2 is usually undetectable but rapidly inducible by a variety of stimuli. COX2 overexpression has been detected in various tumors such as colon, gastric, breast, prostate and lung carcinomas. Up-regulation of COX-2 has been associated with resistance to apoptosis, angiogenesis, tumor invasiveness and poor prognosis. Deletion of COX-2 and selective COX-2 inhibitors can prevent lung cancer growth suggesting a promising therapeutic approach. Although a number of studies on COX-2 expression in NSCLC have been published, little is known on its expression in other types of lung carcinomas such as small cell carcinoma (SCLC) which is characterized by aggressiveness and chemoresistance. In this context we investigated the expression of COX-2 and its cellular and subcellular distribution in carcinomas of neuroendocrine origin such as SCLC and LCNEC (large cell neuroendocrine carcinoma). We also compared the COX-2 tumor expression levels to those of normal lung tissue obtained from the same patients.MATERIALS AND METHODS: Retrospective study of 90 cases of sporadic lung carcinomas of neuroendocrine origin (63 SCLC, 1 SCLC combined with adenocarcinoma component, 26 large cell neuroendocrine carcinoma LCNEC) obtained by bronchoscopic biopsies or FNBs and tumor or lung lobe resections. IHC detection of COX-2 protein expression using anti-COX-2 monoclonal antibody (clone COX 229, 1:100 dilution) and semiquantitative assessment of the expression levels.RESULTS AND CONCLUSIONS: A. In normal lung cells COX-2 expression was absent and rarely weak B. COX-2 expression in carcinomas was mainly cytoplasmic as follows : SCLC (N=59, ND=4) 28.8% with 3+ immunostaining, 25.42% with 2+, 30.5% with 1+ and 15.25% absence of expression, SCLC combined 1/1 with 3+ expression and LCNEC 30.77% displayed 3+ immunoreactivity, 11.54% moderate, 46.15% weak and 11.54% no expression. No significant tumor cell COX-2 expression heterogeneity was observed. In conclusion COX-2 expression was higher in tumor cells compared to normal ones suggesting a putative role of COX-2 in SCLC and LCNEC tumor progression and a promising therapeutic intervention by using selective COX-2 inhibitors against neuroendocrine lung tumors. PP4-105 IMMUNOHISTOCHEMICAL ANALYSIS OF VASCULAR ENDOTHELIAL GROWTH FACTOR SECRETIVE ISOFORMS VEGF121, VEGF165 AND VEGF189 EXPRESSION PATTERN IN SPORADIC SMALL CELL AND LARGE CELL NEUROENDOCRINE LUNG CARCINOMAS Christos Valavanis, Maria Lekarakou, Athanasios Simoulis, Maria Britsou, Maria Terzi, Gino Vecchini, Petroula Arapantoni-Dadioti Molecular Pathology Unit, Dept of Pathology, METAXA Cancer Hospital, Piraeus, Greece BACKGROUND-OBJECTIVE: Angiogenesis is an essential process in tumor progression A number of interacting factors, such as growth factors, receptor tyrosine kinases, matrix metalloproteinases and integrins are involved in this

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phenomenon. VEGF is an important regulator of angiogenesis and its gene gives rise in six protein isoforms through alternative splicing. VEGF exhibits proliferatve action mainly restricted to endothelial cells, stimulates microvascular leakage and induces proteases expression in endothelial cells contributing to tumor invasion and metastasis. High levels of VEGF expression correlate with poor prognosis in a variety of cancers including lung cancer especially NSCLC. However, information on VEGF expression in other lung tumor types such as SCLC is limited and needs further investigation taking into account the high metastatic potential of this carcinoma. In this regard we examined the expression of VEGF isoforms 121, 165 and 189 and their cellular and subcellular distribution in sporadic small cell (SCLC) and large cell neuroendocrine (LCNEC) lung carcinomas.We also compared the VEGF tumor expression levels to those of normal lung tissue obtained from the same patients. MATERIALS AND METHODS: Retrospective study of 90 cases of sporadic lung carcinomas of neuroendocrine origin (63 SCLC, 1 SCLC combined with adenocarcinoma component, 26 large cell neuroendocrine carcinoma LCNEC) obtained by bronchoscopic biopsies or FNBs and tumor or lung lobe resections. Simultaneous immunohistochemical detection of three VEGF isoforms expression using anti-human VEGF monoclonal antibody (clone SP28 1:100 dilution) that reacts with the VEGF121, VEGF165 and VEGF189 splice variants of VEGF. Semiquantitative assessment of the expression levels. RESULTS AND CONCLUSIONS: A. In normal lung cells VEGF expression was absent and rarely very weak . B. VEGF expression in carcinomas was cytoplasmic as follows : SCLC (N=56, ND=6) 3.57% with 3+ immunostaining , 8.93% with 2+, 30.35% with 1+ and 57.14% absence of expression, SCLC combined 1/1 with moderate 2+ expression and LCNEC (N=24, ND=2) 4.17% displayed 3+ cytoplasmic immunoreactivity, 12.5% moderate, 37.5% weak and 45.83% no expression. No significant tumor cell VEGF expression heterogeneity was observed. In conclusion VEGF expression levels in SCLC and LCNEC tumor cells were low or negative and demonstrated no significant difference from normal lung epithelia. This finding may suggest that other angiogenic factors than VEGF are responsible for the high microvessel density observed in these neoplasms. PP4-106 IMMUNOHISTOCHEMICAL STUDY OF EMBRYONIC-LETHAL ABNORMAL VISION-LIKE ELAV/HUR PROTEIN EXPRESSION PATTERN IN SPORADIC SMALL CELL AND LARGE CELL NEUROENDOCRINE LUNG CARCINOMAS Christos Valavanis1, Maria Lekarakou1, Maria Terzi1, Rebeca Kaltsa1, Joanna Iakovidou1, Petroula Arapantoni-Dadioti1, Dimitrios Kontogiannis2 1 Molecular Pathology Unit, Dept. of Pathology, METAXA Cancer Hospital, Piraeus, Greece 2 Dept. of Immunology, Alex. Fleming Biomedical Research Institute, Vari, Attiki, Greece BACKGROUND-OBJECTIVE: Embryonic lethal abnormal vision (ELAV)-like protein HuR belongs to the RNA recognition motif (RRM) protein superfamily and is involved in cell growth and differentiation through posttranscriptional regulation of RNA transcripts. HuR is implicated in the stability and translation of various mRNAs, such as those encoding oncogenes, angiogenic factors and immunosuppressive cytokines. This transcripts stabilization occur via HuR binding to AU-rich elements (ARE) in the 3 untranslated regions (UTR) of mRNAs. Therefore, dysregulation of HuR expression through putative genetic alterations or posttranscriptional and/or translational changes might contribute to malignant tumor progression . Indeed, there is a growing body of evidence that HuR is overexpressed in brain tumors, breast, ovary, NSCLC and colon carcinomas. Thus, HuR could provide a target for new therapeutic approaches by altering

post-transcriptional events of tumor cells. In this context we investigated the expression of HuR and its cellular and subcellular distribution in carcinomas of neuroendocrine origin. We also compared the HuR tumor expression levels to those of normal lung tissue obtained from the same patients. MATERIALS AND METHODS: Retrospective study of 90 cases of sporadic lung carcinomas of neuroendocrine origin (63 SCLC, 1 SCLC combined with adenocarcinoma component, 26 large cell neuroendocrine carcinoma LCNEC) obtained by bronchoscopic biopsies or FNBs and tumor or lung lobe resections. Immunohistochemical detection of HuR protein expression using anti-HuR monoclonal antibody (1:500 dilution) and semiquantitative evaluation of the expression levels. RESULTS AND CONCLUSIONS: A. In normal lung cells HuR expression was both nuclear 2+ and cytoplasmic 1+ immunostaining. B. HuR expression in carcinomas showed : SCLC 87.3%, SCLC combined 1/1 and LCNEC 92.3% displayed 3+ nuclear and 2+ cytoplasmic immunoreactivity . All tumor types were characterized by significant tumor cell HuR expression heterogeneity (almost 30-40% of the tumor cells had no expression at all). In conclusion HuR expression levels were higher in tumor cells compared to normal ones suggesting a putative role of HuR in carcinogenesis and tumor progression via oncogene mRNAs stabilization.An important feature was the high tumor cell percentage (30-40%) with no HuR expression at all suggesting a dysregulation of post-transcriptional machinery leading to putative higher mRNAs degradation rates of other important molecules such as oncosuppressive gene transcripts. PP4-107 PRIMARY PULMONARY MENINGIOMA: REPORT OF A CASE AND REVIEW OF THE LITERATURE Sevinc Hallac Keser1, Sezer Gezgin1, Hakan Karabulut1, Nimet Karadayi1, Benan Caglayan2, Bulent Arman3, Buge Oz4, Philip Cagle5, Enrique M. Gomez5 1 Dr. Lutfi Kırdar Kartal Educational and Research Hospital, Pathology Department, Istanbul, Turkey 2 Dr. Lutfi Kırdar Kartal Educational and Research Hospital, Thorax Diseases Department, Istanbul, Turkey 3 Dr. Lutfi Kırdar Kartal Educational and Research Hospital, Thorax Surgery Department, Istanbul, Turkey 4 Istanbul University Cerrahpasa Medical Faculty Pathology Deparment, Istanbul, Turkey 5 The Methodist Hospital Houston, USA Background: Meningiomas are well recognized in the central nervous system and comprise 15% of all intracranial tumors. Primary ectopic meningiomas are very rare neoplasms which are usually found in the head and neck region (scalp, orbit of the eye, temporal bone, sinonasal area, mandible and ear, paraspinal soft tissues) and less frequently in the mediastinum or retroperitoneum. Primary pulmonary meningioma is very rare. According to the recent literature, there are around 30 cases reported. Gross, histological and immunohistochemical features are similar to their intracranial counterparts. Although in some reports there is slight female predominance, other authors proposed that there is no predilection for sex, age and any particular lobe or segment of lung. These tumors usually present as a solitary pulmonary nodule, grow slowly, are benign in nature and have excellent prognosis. Clinically they can mimic any other pulmonary tumor including metastasis. Case report: A 34 year old woman was accepted to the Thoracic Surgery Department of our hospital with a four months history of cough, dyspnea and occasional hemoptysis. Flexible bronchoscopy revealed a mass lesion in the right lower truncus intermedius blocking bronchial lumen. Atelectasis at the right basal area was discovered with the chest x-ray. With the computed tomography of thorax, right lower atalectasis and intrabronchial solid mass lesion 5.3x3cm in diameters in the right medial-lower lobe was found. Right lung- medial and lower bilobectomy was performed with the clinical diagnoses of bronchial carcinoma or carcinoid

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tumor. Gross specimen of lung tissue revealed a 2.5x2x2cm, grey-white, relatively well circumscribed, hard tumoral lesion blocking the intermediary bronchus lumen. Histologically the tumor was composed of cells with eosinophilic cytoplasm and round to oval nuclei in syncytial arrangement. Occasionally nuclear inclusions, rare mitosis and atypia were observed. Immunohistochemical examination showed positive staining for EMA (epithelial membrane antigen) and Vimentin. Stains for PanCytokeratin, Chromogranin, Synaptophysin, Neuron Spesific Enolase, HHF-35, S-100, HMB45, Desmin, CD68 , GFAP, TTF-1 and CD34 were negative. Conclusion: We present a case of primary pulmonary meningioma by reviewing the literature and discussing the differential diagnosis of this rare neoplasm. PP4-108 THE ROLE OF IMMUNOHISTOCHEMISTRY IN DISTINGUISHING MALIGNANT MESOTHELIOMA AND ADENOCARCINOMA Emine Bagir, Derya Gumurdulu, E. Handan Zeren Cukurova University, Faculty of Medicine, Department of Pathology, Adana, Turkey Background: Histological diagnosis of malignant mesothelioma (MM) and differentiation from adenocarcinoma (AC) is often difficult. In recent years, a large number of immunohistochemical markers have been increasingly applied for the differential diagnosis for these tumors. The aim of this study is to compare the value of MOC 31, HBME-1 and calretinin for discriminating between these malignancy. Materials and Methods: Eighty six cases of MM and twenty six cases of AC retrieved from the files of pathology department of Cukurova University Hospital were included in this study. Immunohistochemistry was performed on each case using avidin-biotin-peroxidese tecnique with antibodies to keratin, EMA, CEA, MOC-31, HBME-1 and calretinin. Results: Keratin was positive in all cases. EMA stained 76 of 86 MM (88.3%) and 23 of 26 AC (88.4%). CEA was positive in 23 of 26 AC (88.4%) and stained 17 MM cases (19.7%). MOC 31 stained 19 of 26 AC cases (73%) as well as 8 of 86 MM (9.3%). HBME-1 was positive 75 of 86 MM (87.2%), thick and membranous staining was prominent. HBME-1 stained 14 of 26 AC (53.8%). Calretinin was positive in %86.3 MM cases, both nuclear and cytoplasmic staining were noted. None of AC cases reacted for calretinin. Conclusion: This study confirms the importance of an immunohistochemical panel in the diagnosis of MM, and the important role of calretinin in this panel. A panel of four markers (two positive and two negative) usally allows for the distinction to be made between MM and AC. Calretinin, MOC 31 and CEA should be included in this panel. PP4-109 ASSESSMENT OF THE PROTECTIVE EFFECT OF VITAMIN D ON RADIATION-INDUCED LUNG INJURY Ferda Bir1, Bahar Baltalarli2, Gulcin Abban3, Nese Demirkan1 1 Pamukkale University, School of Medicine, Department of Pathology, Denizli, Turkey 2 Pamukkale University, School of Medicine, Department of Radiation Oncology, Denizli, Turkey 3 Pamukkale University, School of Medicine, Department of Histology, Denizli, Turkey BACKGROUND: Vitamin D, especially its most active metabolite 1,25-dihydroxyvitamin D3 (calcitriol) is essential in regulating a wide variety of biologic processes, such as blocking mesangial cell activation. Recent studies indicate that calcitriol induces anti-fibrotic hepatocyte growth factor expression, which in turn blocks the myofibroblastic activation and matrix production in interstitial fibroblast. Myofibroblasts express alpha-smooth muscle actin, are present in the early phase of acute lung injury. The objective of this study was to assess the histopathological evaluation of the effectiveness of vitamin D as a protectant against radiation-induced lung injury.

METHODS: The study included 18 Wistar female albino rats with an average weight of 250-300gr. All animals were divided into tree groups. Group 1: control (n=5), Group 2: irradiation alone (n=7), Group 3:irradiation and vitamin D via IM (n=7). Rats were exposed to 20 Gy radiation to the right lung in a Co60 radiotherapy machine where group 3 animals were treated with single dose of vitamin D3 (0.2mgr) injected I.M. for 2 hours before exposure of irradiation. Fifty days after post-irradiation rats were sacrified. The lungs were dissected and blinded histopathological evaluation was performed to asses for lung injury. Immunohistochemically we were stained alpha-smooth muscle actin to highlighted myofibroblasts. RESULT: Light microscopy of right lung sections fifty days after radiation revealed widespread distortion of architecture, with areas that showed markedly thickened alveolar walls. There was abundant vasculitic changes and extravasated neutrophils. Irradiation and vitamin D group of right lung sections also showed thickened alveolar walls but these finding focal rather than diffuse. No vasculer change determined of this group. Histopathologically the lung injury score was significantly lower for irradiation alone when vitamin D was l be needed to prove its safety. administered (p 0.05). Also myofibroblastic differantiation score was significantly lower for the irrradiation alone than radiation and vitamin D group (p 0.05). CONCLUSION: This study indicates that administration of vitamin D3 has an histological evidence of effectiveness as a protectant againts acute lung injury through blocking myofibroblastic activation. Further trials will be needed to prove its safety. PP4-110 ADRENALINE ATTENUATES THE ACUTE LUNG INJURY AFTER INTRATRACHEAL LIPOPOLYSACCHARIDE INSTILLATION: AN EXPERIMENTAL STUDY George Agrogiannis1, Andreas Lazaris1, George Philippakis2, Christos Zissis2, Georgia Thomopoulou1, Penelopi Nikolopoulou-Stamati1, John Bramis3, Despina Perrea4, John Bellenis2, Sofia Tseleni1, Efstratios Patsouris1 1 National and Kapodostrian University of Athens, School of Medicine, 1st Dept of Pathology, Athens, Greece 2 Department of Thoracic and Cardiovascular Surgery, Evangelismos General Hospital, Athens, Greece 3 University of Athens 1st Propaedeutic Surgical Clinic, School of Medicine,National and Kapodistrian University of Athens,Greece 4 Institute of Experimental Surgery and Surgical Research, School of Medicine, National and Kapodistrian University of Athens, Greece Background Endotoxin is a major cause of endotoxinemia, sepsis and pneumonia due to gram-negative bacteria as well. Experimental endotoxin administration via the tracheal route has been extensively used to study the biological and pathophysiologic pathways of inflammation. In particular, experimental endotoxin instillation in the respiratory tree has allowed an extended research with regard to the local response of the lungs to the pathogenic stimulus. Method This study aims to examine the effects of adrenaline subcutaneous infusion in experimental lung inflammation by administration of intratracheal lipopolysaccharide (LPS) in an in vivo animal model. Two groups of animals were used for that purpose, a control group (single LPS administration) and a study group (subcutaneous adrenaline infusion following LPS administration). Results We found that in both groups, a mixed monocytic and lymphocytic infiltrate is an early event during the course of inflammation, occurring approximately two hours post-endotoxin instillation. In the study group, we determined that adrenaline mediated the lung inflammation in a statistically significant degree. By the use of immunohistochemistry, we also identified an increased population of CD4 T-lymphocytes in the inflammatory infiltrate, further endorsing the hypothesis that T-helper lymphocytes, along with monocytes, secrete cytokines

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which amplify the inflammatory response. Conclusion Our study establishes that systemic adrenaline administration after LPS instillation may ameliorate the inflammatory lung response in vivo. PP4-111 LARGE CELL LUNG CARCINOMA –(LCLC)– APPROACH TO RECLASSIFICATION ACCORDING TO THE DEFINITION AND CRITERIA CHANGES IN 1999 AND 2004 WHO CLASSIFICATIONS Cezary Jochymski, Robert Koktysz, Piotr Wi niewski, Wojciech Kozlowski Pathology Dept. Military Institute of Health Services Warsaw, Poland In the Pathology Department of Military Institute of Health Services in the years 1996 to 2007 911 cases of Lung Non Small Cells Cancers have been observed. In this group 89 have been primarily diagnosed as Large Cells Lung Cancers according to definitions and criteria described in previously published WHO Lung Tumors Classifications. The new WHO Classification of Lung Tumors published in 2004 changes some of the morphological criteria and definitions especially in that group of Lung Cancers. Aim: The aim of this study is to reclassify observed and primarily diagnosed Large Cell Lung Cancers according to the changes in current classification using standard microscopy evaluation and histochemical or immunohistochemical methods. Methods: All 89 LCLC cases identified (54 postoperative, 28 from oligobiopsy and 6 interoperative frozen sections) have been reclassified independently by 3 pathologists experienced in pulmonary pathology – selected slides from all cases have been stained with mucicarmin, PaS and alcian blue, CKAE1-3, CK7, CK20, TTF-1, ChromograninA, Synaptophisin, Calretinin, Vimentin and Ki-67/MIB-1 antibodies. Results: From 89 selected LCLC 79,5% cases have been classified as variants of Large Cell Cancer (classic type 44,3%; basocellular 11,3%, neuroendocrine 12,5%, other subtypes 11,4%). 18 (20,5%) observed cases have been classified according to the new WHO criteria and definitions as: Adenocarcinoma – 8%, squamous carcinoma – 10% and sarcomatoid carcinoma – 2,5%. Statistical analysis of Clinical Stage Tumor size, Nodal involvement (pTNM) and some of epidemiological data have also been performed. There were no statistical differences in tumor size (pT), nodal involvement (pN) and observed Pathological Stage between newly classified LCLC and other cancers. Conclusions: New definitions and criteria of 2004 WHO classification change about 20% of previously made diagnoses of LCLC based on earlier editions of WHO Lung Cancer Classifications. It should be emphasised that in our own material about 12% of reclassified LCLC have an neuroendocrine differentiation identified by immunohistochemical methods. For exact diagnosis based on contemporary classification of the LCLC immunohistochemical methods are necessary. PP4-112 EXPRESSION OF SP-B (M-PR) AND TTF-1 IN ATYPICAL ADENOMATOUS HYPERPLASIA (AAH), BRONCHIOLOALVEOLAR CARCINOMA (BAC) AND PULMONARY ADENOCARCINOMA Rodoula Tringidou1, Anna-Maria Athanasiadou1, Katerina Tsouri2, Athanasios Krasas3, Evangelos Sepsas3 1 Dept. of Histopathology, General Athens Hospital of Thoracic Diseases, Greece 2 Dept. of Pneumonology, General Hospital of Leukadas, Greece 3 1st Thoracic Surgery Dept., General Athens Hospital of Thoracic Diseases, Greece BACKGROUND: Atypical adenomatous hyperplasia (AAH) is reported as a focal premalignant change accompanying carcinomas in lungs that are not affected by fibrosis. The tumor that AAH most often accompanies is a peripheral

adenocarcinoma. AAH may be also found in association with multiple adenocarcinomas. AIM OF THE STUDY: We examined the expression of surfactant protein B (mature and precursor/SP-BM, SP-BPr) and thyroid trancription factor-1(TTF-1) in two (2) cases of invasive parenchymal type lung adenocarcinomas develops probably from AAH through an intermediate stage of bronchiolalveolar carcinoma (BAC). PATIENTS: CASE 1: A woman patient aged 59ys with clinical history of bronchitis and asthma. For 20ys she has been on ‘’lediol’’ treatment. Symptoms: cough – dyspnoea. Macroscopically: Surgical specimen of upper lobe of left lung with a tumor mass dim 3x2,8x2,5 cm localized 3 cm from the surgical bronchial edge. Microscopically: Mucin-producing invasive adenocarcinoma with multiple foci of AAH and lesion with BAC growth pattern. CASE 2: A woman patient aged 66ys with cough and dyspnoea has been treated with antibiotics after a recent episode of pneumonia with fever. CT and HRCT findings: Mass dim 4X6X2,5 cm on the upper lobe of right lung. FNB: Well differentiated adenocarcinoma type BAC. Macroscopically: Surgical specimen of upper lobe of right lung with a tumorous mass dim 3X2X3 cm. Microscopically: Peripheral BAC with foci of AAH in surrounding lung parenchyma. METHODS: Immunohistochemistry (IMC) was performed using a panel of primary antibodies such as: epithelial markers (CK 18, 19, 7), CEA, surfactant – protein B (M-Pr), thyroid transcription factor (TTF-1). RESULTS: Case 1: CK 18(+), CK 19(+), CK 7(+), CEA focally (+), SP-BM (-), SP-BPr(+), TTF-1(+). Case 2: CK 18(+), CK 19(+), CEA focally (+), SP-BM (-), SP-BPr(+), TTF-1(+). A variability of expression of SP-BM/SP-BPr was detected on cellular level in foci of AAH comparing with lesions of BAC or invasive adenocarcinoma. CONCLUSIONS: (1) The variability of differences in the expression of SP-B (M and Pr) in foci of AAH near BAC or invasive adenocarcinoma, may be helpful in early diagnosis of this type of lung cancer and in the future the use of these markers is expected to have prognostic value. (2) It is not clear whether multiple foci of AAH and adenocarcinomas (BAC or invasive adenocarcinoma) in the same patient represent independent neoplastic foci or probably clonally related to each other. PP4-113 ENDOGLIN (CD105) EXPRESSION IN NON-SMALL CELL LUNG CANCER AND ITS CORRELATION WITH EGFR, CD34 AND OTHER HISTOPATHOLOGICAL PARAMETERS: PILOT STUDY Nurija Bilalovic1, Adela Cimic1, Semir Vranic1, Ilir Bejtovic1, Adisa Chikha1, Faris Gavrankapetanovic2 1 Department of Pathology, Clinical Center of the University of Sarajevo, Bosnia and Herzegovina 2 Department of Surgery, Clinical Center of the University of Sarajevo, Bosnia and Herzegovina Introduction: Endoglin (CD105), a cell-surface glycoprotein, recently has been identified as an optimal indicator of the proliferation of human endothelial cells. In this pilot study, we wanted to explore its presence in non-small cell lung cancer (NSCLC) and its correlation with epidermal growth factor receptor (EGFR), CD34 as well as with other standard histopathological parameters. Material and methods: Thirty-eight samples of NSCLC, diagnosed in period 2005-2006 at the Department of Pathology, Clinical Center of the University of Sarajevo, were randomly selected and retrospectively analysed. 19 squamous cell carcinomas (50%), 15 adenocarcinomas (39.4%) and 2 samples of adenosquamous and large cell carcinomas (5.3%) were stained for endoglin, CD34 and EGFR using monoclonal antibodies. All immunohistochemical results were captured by Olympus Digital Camera and scored by three researchers independently. Mean value was taken as a final score of immunohistochemical analysis. Statistical Package for Social Sciences (Version 11.5, SPSS, Inc., Chicago, IL, USA) was used for statistical analysis. Results: Endoglin expression was negative in 22 samples (57.9%) whereas positive expression was detected

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in 16 cases (42.1%). The mean number of blood vessels stained by endoglin was 17.18 per field (magnification 200x). It was slightly higher in squamous cell compared with adenocarcinomas whereas CD34 expression had the opposite distribution but without significant association in both cases. Endoglin significantly correlated with another vascular endothelial marker, CD34 (p=0.024, Spearman’s Correlation Rank 0.364). Also, significant positive correlation was detected with vascular invasion, previously confirmed on routine HE samples (p=0.008, Spearman’s Correlation Rank 0.423). Endoglin did not correlate with tumor size whereas CD34 inversely did (p=0.039, Spearman’s Correlation Rank –2.076). CD34 inversely correlated with tumor grade (p=0.05, Spearman’s Correlation Rank –0.289). Vascular endothelial markers did not correlate with EGFR whose expression was slightly higher among squamous cell carcinomas compared with other subtypes. Conclusion: Endoglin is reliable marker of neoangiogenesis and its presence was frequently found in NSCLC. It significantly correlated with vascular invasion, important marker of tumor aggressiveness. PP4-114 DIAGNOSTIC CHALLENGE ON FROZEN SECTION: PULMONARY PAPILLARY PNEUMOCYTOMA (SCLEROSING HEMANGIOMA) AND IMPORTANCE OF CLINICAL DATA Nurija Bilalovic1, Adela Cimic1, Semir Vranic1, Zrinka Vidovic1, Klaus Kayser2 1 Department of Pathology, Clinical Center of the University of Sarajevo, Bosnia and Herzegovina 2 UICC-TPCC, Institute of Pathology, Charite, Berlin, Germany We report here a rare case of primary pulmonary papillary pneumocytoma (sclerosing hemangioma), diagnosed in a 45-year old Bosnian female. Well-demarcated, 1.2x1 cm large, soft, partially calcified, multinodular tumor located in right middle lobe was received for intraoperative analysis. The initial diagnosis was metastatic papillary carcinoma. The tumor was composed of polygonal cells with eosinophilic cytoplasm arranged in papillary pattern (CK7 and EMA positive). The stroma was mainly composed of small, round cells (TTF positive). In between the network of small blood vessels was present (CD31 and CD34 positive). Ki-67 staining (MIB1 antibody) revealed low percentage of positive cells (less than 5%). In consultation with the surgeon we found out that the lesion persisted for several years. This information enabled the final diagnosis as well as second opinion since the sample was consulted in another institution. Although this entity is very care, careful examination and clinical history are of critical importance for accurate diagnosis. PP4-115 INFLAMMATORY MYOFIBROBLASTIC TUMOR (IMT) WITH ABNORMAL OCTREOSCAN® POSITIVITY: A MISLEADING CASE Cinzia Giacometti1, Marco Schiavon2, Federico Rea2, Marialuisa Valente1, Elisabeth Brambilla3, Fiorella Calabrese1 1 Department of Medical and Diagnostic Sciences and Special Therapies, University of Padua, Padua, Italy 2 Department of Cardiothoracic Sciences, University of Padua, Padua, Italy 3 Department of Pathology, Centre Hospitalier Universitaire Albert Michallon, Grenoble, France Background: Inflammatory myofibroblastic tumor (IMT) is a relatively uncommon lesion, which has been reported to occur in nearly every site in the body. We present an unusual clinical presentation of pulmonary IMT, clinically misdiagnosed as carcinoid due to Octreoscan® abnormal positivity. Case Report: In August 2005 a 52 year-old woman, former smoker of 10 cigarettes/day, was admitted in hospital with thoracic pain. At HRCT the nodule appeared as a focal parenchymal solitary

lesion, sharply circumscribed. The 2-Deoxy-2-[18F]Fluoro-D-Glucose (FDG)-PET/CT scan showed increase in tracer activity of the lung mass, suggestive of malignant lesion. The Octreoscan® showed abnormal accumulation of radioactivity in the left hilar region of the lung. A clinical/radiological diagnosis of carcinoid was postulated. An intra-operative frozen section was done, however the confirmed diagnosis of carcinoid could not be achieved. The tumor was composed of spindle cells with a bland cytology, low nuclear atypia and absence of mitosis. The lesion was regarded as low-grade malignancy. The chosen therapeutic procedure was surgical excision (lobectomy). At histology, the lesion consisted of spindle cells arranged in storiform pattern and an admixture of inflammatory cells, with a predominance of hystiocites and lymphocytes. The tumor was negative for ALK1, cytokeratin, CD34, S100 protein and EMA. The inflammatory cells infiltrating showed a strong staining for somatostatin receptors (SSTRs; subtypes 2A and 5). A final diagnosis of IMT was made. Six months after the surgical excision, the patient is alive and well. Conclusion: SSTRs have been identified on human immune cells, including mononuclear leucocytes and peripheral blood lymphocytes. The positive finding of the Octreoscan in our case was due to large intra-tumoral infiltration by lymphocytes and monocyte which expressed SSTRs. To our knowledge, this is the first case of IMT documented with Octreoscan and with immunohistochemistry for SSTRs. PP4-116 ACUTE REJECTION IN HUMAN LUNG TRANSPLANTATION: A POSSIBLE SYNERGIC ROLE OF RHINOVIRAL-CYTOMEGALOVIRUS CO-INFECTION Cinzia Giacometti1, Francesca Lunardi1, Kim Oliani2, Giuseppe Marulli2, Monica Loy2, Federico Rea2, Claudio Schiraldi2, Marina Saetta2, Fiorella Calabrese1 1 Department of Medical and Diagnostic Sciences and Special Therapies, University of Padua, Padua, Italy 2 Department of Cardiothoracic Sciences, University of Padua, Padua, Italy Background: Lung transplantation offers a chance of an improved quality of life and survival for many patients with advanced lung disease. Cytomegalovirus (CMV) and non-CMV viral respiratory infections often involve the lower respiratory tract and are associated with significant morbidity and mortality. The aim of the study was to assess the frequency of different viruses in bronchoalveolar lavage (BAL) specimens and to investigate a potential impact of the most frequent infections on allograft function. Methods: 240 monitoring BAL samples from 48 consecutive lung-transplant patients were analysed by polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR for Cytomegalovirus (CMV), Adenovirus, Rhinovirus (RN), Herpes Simplex Virus, Influenza Virus and Epstein-Barr Virus. Different clinical and morphological parameters, including acute rejection (AR), were recorded for a mean follow up time of 2.86 1.24 years. Results: High frequency of viral infections was detected in our BAL samples (160/240, 67%): particularly CMV and RN (122/160, 76% and 39/160, 24% respectively). Viral co-infections were found in 43/160 (27%) with frequent occurrence of CMV/RN (23/43, 53%). Patients with RN and RN/CMV positive BALs (group 1) were compared with patients with only CMV positive BALs (group 2) and with patients with CMV/other viruses (group 3). Group 1 showed a significant higher rejection index (n ARs/ n total biopsies) than group 2 (p=0.04). Recurrent infective episodes were more frequently detected in group 1 than in group 2 (p=0.006). No significant correlations were observed between group 2 and group 3. Conclusion: 1) CMV/RN is a frequent co-infection in lung transplant patients; 2) recurrent episodes of CMV/RN co-infection and the synergic action of the two viruses could play a crucial role in the induction of AR.

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PP4-117 OXALOSIS AND NECROTIZING PULMONARY ASPERGILLOSIS: TWO CASES Châari Chiraz, Makni Saloua, Chtourou Imen, Fakhfakh Ines, Gouiaa Naourez, Makni Saloua, Khabir Abdelmajid, Sellami Boudawara Tahya EPS Habib Bourguiba SFAX, Tunisia Introduction: The pulmonary oxalosis is a very rare pseudotumoral lesion; it is often secondary to an infection by Aspergillus niger and more rarely by Aspergillus flavus, Aspergillus fumigatus, Aspergillus luchurensis or by Beauveria bassiana. The diagnosis was based on histological examination Our aim is to discuss the diagnosis and the histogenesis of this pathologic association. Observations: We report two cases of chronic necrotizing pulmonary aspergillosis associated with oxalosis among two 17 and 69 year old men with a history of tuberculosis. The thoracic radiography revealed an hydroaeric level in the leg. The computed tomodensitometry revealed a pyopneumothorax and a collapses of the homolateral leg with tuberculosis lesions probably associated to an aspergillosis greff. The serology of aspergillosis was positive in the two cases. The histological analyse of the surgical specimans highlighting the birefringent calcium oxalate crystals by polarisation associated with branching septate hyphae. The culture yielded Aspergillus niger for the young patient. Conclusion: The oxalosis indicates evry intra-tissue oxalate crystal precipitation of calcium hereditary or acquired. Several authors described a brocho-pulmonary deposits of oxalate of calcium associated to an aspergillosis, the oxalic acid is a mycotoxin released especially by A.niger The diagnosis is based on mycologic examination with culture. The treatment is surgical in case of serious hemoptysy or a resistant surinfection to the medical treatment or to the drainage. The evolution is in general favourable. PP4-118 EFFECTS OF PROPOLIS IN THE PROGRESSION OF URETHANE-INDUCED LUNG EXPERIMENTAL CANCER Tamara Veiga Faria, Maristela Munhoz, José Antonio Cordeiro, Patricia Maluf Cury Faculdade de Medicina de Sâo José do Rio Preto, Brazil Background : Propolis, a natural beehive product has been known for centuries for a variety of beneficial traditional medicinal properties. The present study was conducted to ascertain the antineoplastic potential of propolis Design: 3mg/kg urethane was injected in male Balb-c mice, with 7 to 13 life weeks. One group with 13 mice was the control, and another group with 18 mice were submitted to 1.5 mg of Propolis diluted in 350 ml of drinking water . Food and water were given ad libitum. After 16 weeks, all animals were sacrificed and lung nodules and hyperplasias were counted in histological analysis. Results: There was no difference in the number of nodules and hyperplasias between the groups and the control (p= 0.69 and p=0.92, respectively). The final animal weight and food and water ingestion was similar in all groups. Conclusion: Propolis seems to have no effect in the progression of lung cancer in the dose used. It should be interesting to test this substance in higher doses. PP4-119 SCLEROSING THYMOMA: CASE REPORT OF A RARE SUBTYPE Isil Yildiz1, Rıza Dogan2, Pınar Firat1 1 Hacettepe University, Faculty of Medicine, Department of Pathology, Ankara, Turkey 2 Hacettepe University, Faculty of Medicine, Department of Thoracic Surgery, Ankara, Turkey Sclerosing thymoma is classified as a rare subtype of thymoma in the WHO classification. Few reports of this entity have been

made since its first description by Kuo in 1994. Here, we report a case of sclerosing thymoma in a myasthenia gravis patient. A 41 year-old male presented with dysphagia, ptosis and muscle weakness. Computerized tomography showed a 3,5x1,5 cm mass in the anterior mediastinum. With a preliminary diagnosis of thymoma, the patient underwent maximal thymectomy. The specimen was 18x6,5x3 cm in diameter and weighed 175 grams together with the surrounding adipose tissue. On cut surface of the thymectomy specimen a hard, homogenous, spicular mass measuring 4,5x3,5x1 cm and gray-tan in color was detected. Microscopically, the lesion was mostly composed of hyalinized collagen-rich stroma in which aggregates of spindle and polygonal epithelial cells admixed with lymphocytes were noticed. The epithelial cells showed neither atypia nor mitosis. No hemorrhage or necrosis was present. The neoplastic cells were found to be positive for pancytokeratin and negative for LCA and chromogranin. A diagnosis of sclerosing thymoma was made. It is claimed that sclerosing thymoma is occuring as the result of ancient/regressive changes in conventional thymomas. It is a rare and contentious entity which should be considered in the differential diagnosis of small mediastinal biopsies composed of fibrous tissue. PP4-120 INVOLVEMENT OF APOPTOSIS IN AMIODARONE INDUCED PULMONARY TOXICITY AND FIBROSIS Eftihia Kapatou1, Angelos Skyrlas1, Christianna Zachariou1, Konstantinos Hatzistergos1, Theofilos Koletis2, Agathoklis Tsatsoulis3, Maria Bai1, Vassiliki Malamou-Mitsi1 1 University of Ioannina, Medical School, Department of Pathology, Greece 2 University of Ioannina, Medical School, Department of Cardiology, Greece 3 University of Ioannina, Medical School, Department of Internal Medicine-Endocrinology, Greece Background Amiodarone (AMD) is a potent anti-arrhythmic drug frequently used for treating ventricular and supraventricular arrhythmias. Despite its effective antiarrhythmical properties, the use of AMD is often limited by its toxic side effects. The most severe adverse effect of the drug is the Amiodarone-Induced Pulmonary Toxicity (AIPT), which includes alveolitis, phospholipidosis and irreversible fibrosis. However, the mechanism underlying the involvement of AMD to the pathogenic molecular machinery of the disease remains unclear. Recent studies suggest that apoptosis of alveolar epithelial cells (AECs) plays an important role in AIPT. In the present study we investigate the level of involvement of AMD to the triggering of apoptosis. Method The immunohistochemical expression of pro-apoptotic proteins, activated caspase-3 and apoptosis inducing factor (AIF), and anti-apoptotic proteins Bcl-2, survivin and c-Flip, was studied in 33 samples of pulmonary tissue from Wistar rats after oral administration of AMD for 14 days at a dose of 30 mg/kg/day (minimum therapeutic dose). Tissue samples from rats where the drug had not been administered served as controls. Apoptosis was detected by the TUNEL technique and the apoptotic index (AI) was calculated. Results Administration of AMD was correlated with increased apoptosis, increased expression of activated caspase-3, and decreased expression of Bcl-2, survivin and c-Flip. Specifically, we found that AMD had been administered to the 66.7% and 63.2% of samples that presented increased AI and increased expression of activated caspase-3, respectively. On the contrary, administration of AMD was significantly correlated with decreased expression of Bcl-2, survivin, and c-Flip (p 0,001, p 0,001, and p=0,023 respectively). AIF protein showed consistent expression (58.1% of samples) especially among the cellular pulmonary population located far from lymphocyte infiltrations, however did not exhibit any significant correlations. A significant correlation was also found between immunoexpression of activated caspase-3 and TUNEL positive AECs (p=0.040). The immunoexpression

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pattern of Bcl-2, survivin, and c-Flip were also found to be significant correlated (Bcl-2 vs survivin and c-Flip p 0,001 and p 0,001 respectively, survivin vs c-Flip p 0,001). Conclusion Apoptosis plays a crucial role in AIPT. In contrast to AIF protein, activated caspase-3 acts as the effector molecule of apoptosis in AIPT. Inhibition of the anti-apoptotic molecules Bcl-2, survivin, and c-Flip is also involved in AIPT. PP4-121 EXPRESSION OF SMAC/DIABLO AND CASPASE-3 IN NON SMALL CELL LUNG CARCINOMA (NSCLC) Nalan Akyurek1, Ahmet Selim Yurdakul2, Pelin Borcek1, Sedat Demircan3, Cuneyt Kurul3, Can Ozturk2, Leyla Memis1 1 Gazi University Medical School, Department of Pathology, Ankara, Turkey 2 Gazi University Medical School, Department of Pulmonary Diseases, Ankara, Turkey 3 Gazi University Medical School, Department of Thoracic Surgery, Ankara, Turkey Background: The second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein with low pI (Smac/DIABLO) was recently identified as a protein that is released from mitochondria in response to apoptotic stimuli and promotes apoptosis by antagonizing inhibitor of apoptosis proteins. Caspase-3 is a cysteine protease that plays an important role in the process of apoptotic cell death. However, the involvement of Smac/DIABLO and caspase-3 in NSCLC is not fully understood. Method: Expressions of Smac/DIABLO and caspase-3 were investigated immunohistochemically in 112 non-small cell lung carcinoma patients. Staining patterns were correlated with tumor histological type, TNM stage and prognosis. Results: Smac/DIABLO was detected in 12 (11%) and caspase-3 in 60 (53%) of 112 NSCLC cases. There was no significant relationship between Smac/DIABLO and caspase-3 expression and age, gender, histological type, TNM stage and prognosis. In addition, no significant correlation was found between Smac/DIABLO and caspase-3 expression. Conclusion: Our data demonstrate that expression of Smac/DIABLO is rare among NSCLC’s, suggesting that Smac/DIABLO or its agonists is unlikely to be an appropriate target for therapeutic options in patients with these tumors. PP4-122 MYXOID TYPE OF INFLAMMATORY MYOFIBROBLASTIC TUMOR OF THE LUNG: REPORT OF A CASE WITH AN UNUSUAL PRESENTATION Dilek Yılmazbayhan1, Yasemin Ozluk1, Sukru Dilege2, Yusuf Bayrak3, Henry D Tazelaar4 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey 2 Istanbul University, Istanbul Faculty of Medicine, Department of Thoracic Surgery, Istanbul, Turkey 3 VKV American Hospital, Istanbul, Turkey 4 Mayo Clinic, Scottsdale, USA BACKGROUND Inflammatory myofibroblastic tumor (IMT) is a subgroup of inflammatory pseudotumors and has been reported in multiple anatomic locations including lung. It is composed of a variable mixture of collagen, inflammatory cells and a spindle cell proliferation showing myofibroblastic differentiation. IMT occurs mostly in patients less than 40 years. The clinical findings depend on the site of the involvement. Some authors believe that IMT is a reactive process rather than a tumoral lesion, while some support the theory of IMT as being a low grade mesenchymal malignancy. METHODS We report of a case of IMT with an unusual clinical presentation. A 28 year-old man was referred to the hospital with massive hemoptysis. A mass, which was suspicious for arteriovenous malformation, aneurysm or malignancy, was detected in the right upper-lobe adjacent to the interlobar fissur, radiologically. Angiography was in normal

limits. Because of the high risk for hemorrhage, a transthoracic aspiration biopsy was done peroperatively. The sample was generally hypocellular, however there were some cellular areas consisted of mesenchymal cells with vesicular nuclei bearing a centrally located nucleoli. In some cells binucleation was seen. The clinician was warned for a possible mesenchymal lesion which malignant potential could not be predicted. Because the borders of the tumor could not be differentiated from the adjacent parenchyma and the extensive hemorrhagic appearance of the lung, the patient underwent a right upper-lobectomy. RESULTS On macroscopic examination a peripheric mass, that had irregular borders and hemorrhagic cut surface, and measured 3 cm in diameter, was detected in the hilus of the lobe. Histology revealed a well-demarcated tumoral lesion composed of myxoid stroma, neovascularization and bland appearing spindle cells. Binucleated cells, atypia and pleomorphism, rare mitoses were also seen. Immunohistochemistry displayed positive reaction for vimentin, smooth muscle actin, CD31 and p53, whereas no staining was detected for S-100, CD34, desmin, cytokeratin, ALK-1 and HMB-45. The possibility of a metastatic tumor was excluded by PET screening. CONCLUSION IMT is a rare tumor with various histologic patterns that can be confused with other malignant and benign tumors. The diagnosis can be very difficult, especially at the time of surgery in the frozen room. The appropriate choice of treatment is the total excision of the lesion, besides IMT is believed to be a low-grade malignant tumor. Although most of the patients are asymptomatic, clinical presentations differ according to the localization and the components of the tumor. PP4-123 ENDOBRONCHIAL HAMARTOMA: SHOULD THE DIAGNOSIS BE MADE ON A BRONCHOSCOPIC BIOPSY? Yasemin Ozluk1, Levent Tabak1, Sukru Dilege2, Yusuf Bayrak3, Dilek Yilmazbayhan4 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey 2 Istanbul University, Istanbul Faculty of Medicine, Department of Thoracic Surgery, Istanbul, Turkey 3 VKV American Hospital, Istanbul, Turkey 4 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey BACKGROUND Pulmonary hamartomas (PHs) are rare benign tumors composed of varying proportions of mesenchymal tissues and combined with entrapped respiratory epithelium. PHs are usually peripheric, however 3-20% of tumors are endobronchial. Endobronchial lesions clinically cause obstructive symptoms and are localized within the larger airways as broad-based polyps. Histological examination reveals mature cartilage tissue, surrounded by other mesenchymal elements and clefts of respiratory epithelium. In endobronchial hamartomas fat tissue may predominate and epithelial component may be shallow or absent. METHODS We report a case of an endobronchial hamartoma occuring in a 35 year-old man, who was admitted to the hospital with a 2 years of history of recurrent pneumonia and cough. Radiology demonstrated postobstructive pneumonia findings. A bronchoscopy was done with suspicion of a malignancy. Endoscopically, a polypoid lesion measured 0.5 cm in diameter was detected within one of the segment bronchi of right lower-lobe. RESULTS Histologically, bronchoscopic biopsy displayed structures of irregular chondroid and other mesenchymal elements in small tissue fragments . The possibility of a chondroid hamartoma was reported. However a malignant tumor could not be excluded. The patient underwent a segmentectomy procedure. In the frozen section room the mass defined by bronchoscopy could not be detected macroscopically. The bronchoscopy was repeated peroperatively and the mass was seen within an abnormally branching segmental bronchus. The segment carrying the mass was resected. Histologic examination

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of the lesion confirmed the initial diagnosis with chondroid, myxoid mesenchymal and adipose tissue elements containing mass within the abnormal superior segmental bronchus. CONCLUSION PHs are benign lesions which have similar radiological and clinical features with malignant tumors. A biopsy is necessary for the differential diagnosis. Preoperative or peroperative diagnosis is important, since the definitive diagnostic procedure is segmentectomy or lobectomy. The presence of small, irregular chondroid and mesenchymal tissues can be seen in both mesenchymal and hamartomatous tumors. The pathologist and the bronchoscopist have to keep in touch to make the diagnosis. PP4-124 LYMPHOVASCULAR INVASION PREDICTS RECURRENCE IN RESECTED STAGE I NON-SMALL CELL LUNG CARCINOMA Sibel Percinel1, Serpil Dizbay Sak1, Berna Savas1, Serkan Enon2, Ayten Kayi Cangir2 1 Ankara University, Medical Faculty, Department of Pathology, Ankara, Turkey 2 Ankara University, Medical Faculty, Department of Thoracic Surgery, Ankara, Turkey Introduction: Prognosis of patients with non-small cell lung carcinoma (NSCLC) still remains a major problem. Numerous biological markers have been evaluated for prognostic significance but the results have been conflicting. In NSCLC, the prognostic value of lymphovascular invasion (LVI) has been evaluated in early stage patients. Recently, LVI has been reported to be a prognostic factor in patients with resected NSCLC with intrapulmonary metastases. Perineural invasion (PNI) has already been regarded as one of the prognostic parameters to be included for the next staging revision. Materials and Methods: Intratumoral LVI and PNI were investigated in 58 patients who had Stage I (T1N0M0) NSCLC by hematoxylin and eosin stained sections. LVI was determined by the presence of tumor cells either in lymphatic or blood vessels. Survival curves were estimated using the Kaplan-Meier method. The prognostic value of individual clinical and pathological variables was analyzed by the log-rank test. Results: LVI was detected in 50% and PNI was detected in 20.7% of NSCLC cases. The median follow-up for the entire group was 35.5 months. In univariate analysis, neither LVI nor PNI was predictor of overall survival. However, LVI was found to be significantly predictive of recurrence. The estimated relative risk of recurrence in patients with LVI was 3 compared with patients without LVI. Conclusion: LVI predicted recurrence in resected TIN0M0 NSCLC cases. PP4-307 DIAGNOSIS OF DIFFUSE PULMONARY INTERSTITIAL DISEASE BY TRANSBRONSCHIAL BIOPSY (TBB): OUR EXPERIENCE (322 CASES). Ana Puras1, Ana Echegoyen Silanes1, Alfredo López-Cousillas1, Marta Rezola Bajineta1, Eduardo Urbiola Marcilla1, Joan Boldú Mitjans2 1 S. de Anatomía Patológica Hospital Virgen del Camino Pamplona, Spain 2 S. de Neumología Hospital Virgen del Camino Pamplona, Spain Introduction: Transbronchial pulmonary biopsy (TBB) is a good recognized method in the diagnosis of the Lung Interstitial Diseases and its utility has been proven throughout the years. Current technologies (immunohistochemical, molecular and other types of studies) allow a more precise diagnosis. The conditions that are necessary to assess these samples have been simplified in the last years, as well as the technical behavior of them. Also permit to obtain multiples samples of Interstitial Diffuse Diseases in different categories of hospitals. Material and methods: From 1985 to the present moment 322 TBBs have been made in our hospital to patients with suspicion of interstitial Disease, 307 of

which were immunocompetent patients; a meticulous protocol has been followed with all of them for the complete use of the sample: initial seriation of minimal specimens (20 preparations, numbered and occasionally mounted in slides for immunohistochemical techniques) and a later staning, alternatively, of 8 of them: 4 H.E, 1 Masson, 1 PAS, 1 Perls, 1 Blue Alcián; the samples not stained initially, are stained afterwards, if they are necessary; and if not, they are filed. All the cases have been reviewed, in Clinical-Pathological Conferences, and the results of the biopsy have been assesed. They have been always valued with the bronchoalveolar lavage (BAL). Results: The diagnosed pathology has been: A) Sarcoidosis (23), Tuberculosis (4), Hypersensitivity Pneumonitis (30), Eosinophilic Chronic Pneumonia (9), “BOOP pattern” (27), Respiratory Bronchiolitis-Interstitial lung Disease (DIP pattern) (32), Constrictive Bronchiolitis (3), Lung hemorrhage (7, one of them a Goodpasture's Syndrome and other vasculitis), Usual Intersititial Pneumonia (33), Pneumoconiosis (11), Acute lesion (14) (Diffuse Alveolar Damage, Infectious Organizing Pneumonia, Iatrogenic damage,…), Infarct (2), Lymphangioleiomyomatosis. (2), Alveolar Proteinosis (2), Pneumocystis (1), Herpes virus (2), Aspergillosis (2), Bronchoalveolar Carcinoma (9), Lymphoma-Leukemia (4), Carcinomatous lymphangitis (5); B) The diagnosis was orientative in 34; C) Without alterations in 31; and D) The sample was insufficient in 34. Conclusions: 1.- The TBB was diagnostic or orientative, with a latter clinical confirmation in 80% of the cases. 2.- In 20% of the cases it the TBB was insufficient, or non valid (non representative of the process). 3.- The maximum use of the samples and the meticulous technical behavior, as well as the multidisciplinary team work, have contributed to it in a definitive way.

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Skin Pathology PP4-125 DEEP PENETRATING NEVUS, BLUE NEVUS AND SPITZ NEVUS – PROBLEMS OF DIFFERENTIAL DIAGNOSIS Geanina Micu1, Eliza Gramada1, Cristiana Popp1, Alina Stoica1, Alina Georgescu1, Sabina Zurac1, Alexandra Bastian1, Carmen Ardeleanu2, Florica Staniceanu1, Dorina Giurcaneanu3 1 Colentina University Hospital, Department of Pathology, Bucharest, Romania 2 Victor Babes Institute, Bucharest, Romania 3 Colentina University Hospital, Department of Dermatology, Bucharest, Romania Background: Deep penetrating nevus (DPN) is a variant of melanocytic nevus that can be clinically mistaken for malignant melanoma (MM), pigmented nevus (PN), blue nevus (BN) and Spitz nevus (SN). Also, the histopathological features prone to confusion with MM, SN and BN. The final diagnosis is established by histopathologic/ immunohistochemical studies. BN is a benign lesion with two recognized variants: the common blue nevus and the cellular blue nevus. It is believed to represent dermal arrest of neural crest melanocytes during embryonal migration towards epidermis. SN (benign juvenile melanoma, spindle and epithelioid cell nevus) is a benign melanocytic nevus, which shares some histopathological features with malignant melanoma. Material and method: We analyzed the nevocelular tumors diagnosed in our department during two years (2005-2006): 574 nevocelular nevi and 54 MM. There were two DPN, two BN and two SN. We studied the histological features of all tumors (cellularity, cellular polymorphism, mitotic figures and lymphocytic infiltrate), the integrality of resection and the immunohistochemical markers for melanocytic tumors (S100, HMB 45, Melan-A, NSE). Results: Most of the patients were women (5 out of 6), with medium age 39.6 years. Most lesions occurred on the face (4 out of 6), one case on the foot (plantar side), the other having lumbar localization. All the tumors were composed of clusters and fascicles of polygonal and spindle shaped cells, with abundant cytoplasm and bland nuclei, sometimes with visible nucleoli. Cellular and nuclear polymorphism was noted in all cases. Evident diminution in cell size (maturation) towards the base of the lesion was present. Mitoses were present in one DPN (mitotic index 10/100 high power fields), none of them atypical. None of the tumors had pagetoid invasion of the epidermis. The DPNs had additional large, multinucleated tumoral cells in the superficial part. Conclusion: DPN, BN and SN are benign melanocytic tumors, difficult to differentiate both one from the other and especially from a MM. The correct diagnosis has the outmost importance for the patient’s treatment and prognosis. PP4-126 POSITIVE AND DIFFERENTIAL DIAGNOSIS IN TUMORS WITH SEBACEOUS DIFFERENTIATION Florica Staniceanu, Sabina Zurac, Geanina Micu, Alexandra Bastian, Eliza Gramada, Alina Stoica, Alina Slavnea, Ionela Celea Colentina University Hospital, Department of Pathology, Bucharest, Romania Background: Benign sebaceous lesions are frequent, but malignant cutaneous tumors with sebaceous differentiation are relatively uncommon. In this group sebaceous carcinoma is a rare variant of cutaneous tumor with an aggressive biological potential showing a marked tendency for both local recurrence (9-36%) and distant metastasis (14-25%). Method: We studied 18 cases of sebaceous tumors diagnosed in our department in the last four years; 4 of them were sebaceous carcinoma SC, 6 were basal cell carcinomas with sebaceous differentiation (BCCSD), 4 were Jadassohn’s nevi (JN) and 4 were benign sebaceous proliferations (1 sebaceous adenoma (SA), 3 sebaceomas), analyzing

histological patterns, cytoplasmic and nuclear aspects, mitoses, necrosis, invasion and IHC features. Results: There was a significant predominance of male patients (61.1%); the average age differed within the five groups of lesions: 63 for SC, 49.2 for BCCSD, 38.9 for JN and 74.2 for sebaceoma. All the SC were located in the ocular area (3 in the eyelid and one on the conjunctiva); the tumors belonging to the other groups were both ocular and extra ocular (mainly on the scalp). All the SCs were diagnosed as grade II and showed lobular proliferation of atypical vacuolated cells with sebocityc differentiation, necrosis and, in one of the cases, pagetoid invasion of the epidermis; the well differentiated areas showed some similarities with benign sebaceous tumors that are analyzed. The BCCSDs are considered to be borderline tumors as they do not metastasize so that the identification of conventional BCC features associated with mature sebocytic elements were essential to separate them from SC. While the SA revealed proliferation of fully differentiated sebocytes and basaloid cells, the sebaceomas were composed of mixed immature and mature sebocytes. JN presented marked acanthosis, papillomatosis and sebaceous hyperplasia with apocrine proliferation. Histological aspects and IHC data are discussed. Conclusion: Tumors with sebaceous differentiation have various clinical presentations and histopathological aspects that make differential diagnosis both important and difficult. On one hand, well-differentiated sebocytic component of malignant tumors can be mistaken for a benign lesion especially in small bioptic specimens; on the other hand, the BCC component in a malignant tumor with sebocytic differentiation must be separated from SC. Careful examination of numerous areas of the tumor and IHC test must be performed in order to establish the correct diagnosis. PP4-127 IMMUNOHISTOCHEMICAL EXPRESSION OF HUMAN HERPES VIRUS–8 IN THE DIAGNOSIS OF KAPOSI’S SARCOMAS Banu Dogan Gun1, Burak Bahadir1, Mustafa Ozkan Gun2, Erdogan Gonulal2, Neslihan Kokten1, Nilufer Onak Kandemir1, Nimet Karadayi3, Sukru Oguz Ozdamar1 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Zonguldak Ataturk State Hospital, Department of Pathology, Zonguldak, Turkey 3 Dr. Lutfi Kirdar Kartal Research and Training Hospital, Department of Pathology Istanbul, Turkey BACKGROUND: Kaposi’s sarcoma is a low-grade vascular neoplasm that has been shown by molecular analyse to uniformly express latent nuclear antigen-1 of human herpes virus 8. In the present study we assesed the sensitivity and specifity of human herpes virus 8 latent nuclear antigen-1 in the diagnosis of Kaposi’s sarcoma and its potential usefulness for distinguishing it from various mimickers. METHOD: In total, 35 cases of Kaposi’s sarcoma from 25 patients and with 18 mimickers (two cases of spindle cell hemangioendothelioma, three cases of kapiller hemangioma, a case of cavernous hemangioma, two cases of pyogenic granuloma, a case of stasis dermatitis, three cases of fibromatosis, two cases of dermatofibroma, a case of pilar leiomyoma, a case of fibroma and two cases of blue nevus) were included in this study. Fixed, paraffin embedded tissue sections were examined immunohistochemically using human herpes virus 8 latent nuclear antigen-1. Strong diffuse nuclear staining was considered as positive. Also the following variables for Kaposi’s sarcoma cases were analysed; age, sex, localization and stage of the tumor. RESULTS: Of the 25 Kaposi sarcoma’s cases there were 16 males and 9 females, giving an overall male:female ratio of 1.7.The age of the patients ranged from 37 to 80 (mean 66.36) years. The lesions occured more commonly in the lower extremity (54.29%), upper extremity (25.71%) head and neck region (17.14%) and trunk (2.86%). Eight biopsy specimens were in the patch or plaque stage that reflects the early

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stage of Kaposi’s sarcoma and 27 were in nodular stage. In 35/35 cases of the Kaposi’s sarcoma showed positive nuclear staining for human herpes virus 8 latent nuclear antigen-1 (sensitivity, 100%), whereas 3/18 of the mimickers; a case of capillary hemangioma, a case of stasis dermatitis and a case of a dermatofibroma was positive for this antigen (specifity, 83.34%). CONCLUSION: The high sensitivity of human herpes virus 8 in our cases of Kaposi’s sarcoma (100%) were similar with other investigations using immunohistochemistry or polymerase chain reaction. However, human herpes virus 8 specifity of 83.34% detected in this study was a little lower when compared to the literature. The result indicate that its use for diagnostic purpose must be carried out with caution. PP4-128 C-KIT (CD117) EXPRESSION IN CLASSIC KAPOSI SARCOMA Nilufer Onak Kandemir1, Banu Dogan Gun1, Burak Bahadir1, Gamze Yurdakan1, Mustafa Ozkan Gun2, Neslihan Kokten1, Nagehan Ozdemir3, Nimet Karadayi3, Sukru Oguz Ozdamar1 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Zonguldak Ataturk State Hospital, Department of Pathology, Zonguldak, Turkey 3 Dr. Lutfi Kirdar Kartal Research and Training Hospital, Department of Pathology Istanbul, Turkey Background: Kaposi sarcoma (KS) is a multicentric angioproliferative neoplasm characterized histologically by the progressive proliferation of spindle-shaped tumor cells that progresses from patches to plaques, with eventual formation of nodule. Human herpesvirus 8 (HHV8) is associated with all epidemiologic forms of KS and has been shown in vitro to induce the tyrosine receptor kinase c-kit in HHV8- infected endothelial cells. The aim of this study was to evaluate c-kit expression by immunohistochemistry in different proliferative stages of classic Kaposi sarcoma. Method: In total, 30 cases of Kaposi sarcoma; at various histologic stages (4 patch, 2 plage, 24 nodular); from 18 patients were included in this study. Formalin-fixed, parafin- embedded tissue sections were stained using immunohistochemistry with antibodies to c-kit. Strong diffuse cytoplasmic staining was considered as positive. Also the following variables for Kaposi’s sarcoma cases were analysed; age, sex, localization and stage of the tumor. Result: Of the 18 Kaposi sarcoma’s cases there were 12 males and 6 females, giving an overall male:female ratio of 2/1.The age of the patients ranged from 37 to 80 (mean 66.36) years. The lesions occured more commonly in the lower extremity (54.29%), upper extremity (25.71%) and head and neck region (20%). C- kit immunoreactivity of lesional cells was demonstrated in 19 (63.33%) cases. There was no difference in the intensty of c-kit staining between age, sex, localization and histologic stages of KS. Conclusion: These findings indicate that c- kit expression in lesional cells can be detected by immunohistochemistry in different histologic stages of KS. The detection of c-kit expression in KS reveals a potential target for pharmacologic intervention that warrants further study. PP4-129 MAST CELLS AND P53 EXPRESSION IN PSORIASIS VULGARIS Figen Barut, Sibel Bektas, Banu Dogan Gun, Burak Bahadir, Gamze Yurdakan, Sacide Colak, Sukru Oguz Ozdamar Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey Background: Psoriasis vulgaris is a chronic inflammatory T-cell mediated immune dermatosis, characterized by a high epidermal cell turnover, which results in a typical epidermal hyperplasia. As a result of the studies carried on by using antibody of “mast cell tryptase and toluidin blue stain”, which are specific mast cell

determiners, mast cells are found to be increased in psoriasis lesions. In addition, it is observed that there is an increase in cell proliferation of these lesions when p53 is used. The purpose of this study is to show the relationship between p53 protein accumulation that prevents epidermal hyperproliferation and mast cell number in its pathogenesis. Methods: In this study, punch biopsies of the 56 cases with diagnosis of psoriasis vulgaris have been investigated. The presence of mast cells in which the granules stain metachromatic with 1% toluidin blue solution has been investigated in cross-sections of formalin-fixed and parafin-embedded tissue samples. In addition, mast cells that show immunoreaction to mast cell tryptase antibody have been observed by streptavidin-biotin peroxidase technique. p53 proportion (number of positive nuclear cells/1000) in epidermal keratinocytes has been determined. Results: Fifty six cases with psoriasis vulgaris consisted of 31 (55.4%) females and 25 (44.6%) males. The ages of the cases range between 6 and 68. The average age is found 41.25 ± 14.14 (mean ± SD). In psoriasis cases, mast cells number (mean: 61.44± 25.98) and p53 proportion (mean: 454.93± 221.07) are found. In control group, mast cells number (mean: 74,61 ± 25,89) and p53 proportion (mean: 176,36 ± 109,18) were determined. A significant difference between psoriasis cases and control group in terms of mast cell number (p<0.008) and p53 proportion (p<0.001) was detected. No correlation has been found between mast cell number and p53 proportion. However, a moderate degree correlation (r:0,30, p<0.05) has been found between mast cell number and p53 proportion in the control group. Conclusion: Since the obtained correlation between mast cell number and p53 proportion of healthy skin is not valid for psoriasis cases, it is thought that these variables act differently in pathogenesis of psoriasis vulgaris. PP4-130 EXPRESSIONS OF HEREGULIN IN MELANOCYTIC NEVUS, DISPLASTIC NEVUS, AND MELANOMA Aslı Altaykan Hapa1, Gul Erkin1, Ozay Ozkaya2 1 Hacettepe University, Department of Dermatology, Ankara, Turkey 2 Hacettepe University, Department of Pathology, Ankara, Turkey BACKGROUND. Heregulins are natural ligands for the growth factor family receptors called c-erbB-3 and c-erbB-4. Current studies suggest that thay can regulate a variety of responses including proliferation, differentiation, and survival. In addition, overexpressions were shown to be associated with stimulation of carcinogenesis. In this report, we study the expressions of heregulin in melanocytic, displastic nevi, melanoma and investigate their role in neoplastic transformation of melanocytes . METHOD. Using specific antibodies of heregulin, formalin-fixed paraffin embedded specimens of 13 melanoma, 8 displastic nevi, and 12 melanocytic nevi were examined for the expressions of heregulin. RESULTS. Nine of 13 (69%) melanoma, 7 of 8 (87%) displastic nevi, and 8 of 12 (66%) melanocytic nevi tissues showed moderate to strong immunoreactivity. Predominantly diffuse cytoplasmic staining was observed. Junctional melanocytic nests had stronger reactivity than dermal nests. Seventeen out of 33 (51 %) samples expressed heregulin in > 50 % of tumor cells. Additionally, keratinocytes at the upper layer of the epidermis also showed moderate to strong immunoreactivity in all samples. CONCLUSION. Although, heregulin expression were found in the majority of the tissues, we have demonstrated variable expression of heregulin in nevi and melanomas which do not differantiate between nevi and melanomas, but suggest an idea to investigate the heregulin as a new melanocyte marker. As the number of the tissues investigated in our study were taken into consideration, analysis of heregulin expression in larger series will enable us to identify as a useful marker.

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PP4-131 MRNA VEGF-A EXPRESSION AND VEGF-A PROTEIN IN SKIN WOUND HEALING Lovorka Batelja, Bruno Vuleti , Luka Br i , Ivana Ili , Sanja Kapitanovi , Predrag Sikiri , Sven Seiwerth Department of Pathology, Medical Faculty, University of Zagreb, Croatia Introduction: Healing is a complex process of initial inflammation, granulation tissue formation and tissue remodeling. Angiogenesis is a necessary part of wound healing, and VEGF-A is a dominant mediator of physiological and pathological angiogenesis. VEGF-A is a mitogen for endothelial cells. Syntetic pentadecapeptide BPC 157 has so far demonstrated influence on healing process in morphological studies. To our knowledge most of healing studies involving VEGF were performed in vitro. Purpose of the study: to analyze in vivo wound healing by second intention through expression of mRNA VEGF- A and protein VEGF-A and attempt of compare them in control group and group treated with BPC 157. Methods: Full-thickness excisional wounds were made on the back of adults Wistar rats (3 animals / group). Animals were treated with BPC 157; 10 g / kg, i.p. or eqivolume of saline.. Wounds were left uncovered and harvested 4, 6, 12, 24, 48, 72, 120, 168 hrs after injury. Half of the full thickness skin wound area was taken for histology/immunohistochemistry and molecular analysis respectively. For immunohistochemistry we used VEGF 147 antibody (Santa Cruz Biotechnology) following manufacturers protocol. For analysis mRNA VEGF expression analysis RT-PCR method was used. Results: In our study the expression of mRNA VEGF-A in control group followed the published data in wound healing by second intention (VEGF levels peaked at 6 and 48 hrs post-injury) and in group treated with BPC 157 mRNA VEGF showed a sinusoidal curve with three positive and two negative peaks tending toward basal level. Expression of VEGF-A protein was found in epidermis, dermis, hypodermis in both groups of animals, in vessels and in perivascular area. VEGF protein expression was significantly elevated 12 hrs post-injury in vessels of hypodermis in treated group and 48 hrs in perivascular area of hypodermis control group of animals. VEGF protein expression was significantly elevated 168 hrs post-injury in epidermis, and 12 hrs in vessels of dermis in control group. Conclusion: Our data suggest that BPC 157 has influence on mRNA VEGF and protein VEGF-A expression. Treatment with BPC 157 results with better controlled and prolonged expression of VEGF. Control group shows that in vivo wound healing by second intention confirms results of in vitro studies. PP4-132 INFANTILE SYSTEMIC HYALINOSIS, A CASE REPORT Ozge Gunduz1, Sibel Ersoy Evans1, Koray Boduroglu2, Yasemin Alanay2, Ozay Ozkaya3 1 Hacettepe University, Department of Dermatology, Ankara, Turkey 2 Hacettepe University, Ihsan Dogramaci Children 's Hospital, Department of Pediatrics, Clinical Genetics Section, Ankara, Turkey 3 Hacettepe University, Department of Pathology, Ankara, Turkey BACKGROUND. Infantile systemic hyalinosis (ISH) is an autosomal recessive, rare and fatal disease. Hyaline deposition occurs in multiple organ systems, including skin. It is characterised by painful joint contractures, small pearly papules on the head, flesh nodules in the perianal region, gingival hypertrophy, generalized osteopenia, diarrhea, recurrent infections and usually thickened skin. The onset occurs within the first few weeks of life and death occurs by two years of age as a result of recurrent pulmonary infections and diarrhea. Unfortunately, treatment is primarily palliative as there is no cure currently available. METHOD AND RESULTS. Herein, we report an 8-month-old boy that was referred for a rash that

appeared on his neck approximately 4 months earlier. He was born after a full-term pregnancy by caesarean section due to breech presentation. He was the sixth child of non-consanguineous parents who were from the same village. There was no family history of ISH. At 2 months of age, his mother noticed decreased upper and lower limb movements. His physical examination revealed gingival hypertrophy, extensive pearly erythematous papules all around his neck, a 4x4 cm purple plaque located over the sacral area, multiple flesh nodules around perianal region and purple patches located over his right 3th and 4th proximal interphalangeal joints and on both ankles. There was also extension contracture and stiffness in his cervical region, flexion deformity of his limbs with frog-leg position. Radiological evaluation showed generalized osteopenia and metaphysial enlargement. Histopathological examination of one of the papules showed compact amorphous eosinophilic hyaline deposits in the dermis. With the characteristic clinical presentation and histopathological findings, a diagnosis of ISH was made. Unfortunately, the patient died at 16 months of age. CONCLUSION. When encountering an infant with joint stiffness, purplish discoloration over the joints and papules around the neck, ISH should be a differentiative diagnostic consideration. PP4-133 THE VALUE OF IMMUNOPHENOTYPING AND T-CELL RECEPTOR GAMMA GENE REARRENGEMENT IN THE DIAGNOSIS OF MYCOSIS FUNGOIDES Arbil Acıkalin1, lhan Tuncer1, Melek Ergin1, Yasargul Denli2, Gulsah Seydaoglu3 1 Cukurova University, Medical School, Department of Pathology, Adana, Turkey 2 Cukurova University, Medical School, Department of Dermatology, Adana, Turkey 3 Cukurova University, Medical School, Department of Biostatistics, Adana, Turkey Background: Mycosis fungoides is characterized with neoplastic infiltration of skin by dominant CD4 positive T lymphocytes. However, the diagnosis may be difficult for both dermatologist and pathologist in early lesions, hence clinical and histomorphological features may be nonspecific. Our aim was to evaluate the value of immunophenotyping and T cell receptor gene rearrengement studies as an adjunct to the histomorphological diagnosis of mycosis fungoides. Method: Histomorphological, immunohistochemical and polymerase chain reaction analysis were performed on formaline fixed, parafine embedded tissue sections of 73 cases (39 mycosis fungoides with classical histomorphology, 16 with suspicious histology for mycosis fungoides, 18 benign inflammatory dermatoses as a control group). Histomorphologically, three major (epidermotropism, Pautrier microabscess, atypical lymphocyte) and two minor (spongiosis, collagen increase) criterias were evaluated on H&E slides. Antibodies to CD3, CD4, CD8 by using immunohistochemistry and T cell receptor gene rearrengement analysis with polymerase chain reaction were performed. Results: Significant differences were found in epidermotropism (p=0,000), presence of Pautrier microabscess (p=0,004) and atypical lymphocyte (p=0,000), CD4 percentage (p=0,006), CD4/CD8 ratio (p=0,010), and clonality (p=0,000). Besides, CD8 percentage was not statistically different between three groups. In our study, we found out diagnostic cut-off values for CD4 percentage and CD4/CD8 ratio. That was 26% (sensitivity 82%, specifity 69%) for CD4 percentage and 0,65 (sensitivity 74%, specifity 69%) for CD4/CD8 ratio. Conclusion: Histomorphological features are still gold standart for diagnosis of mycosis fungoides. However, in early mycosis fungoides and the suspicious cases, immunophenotypical evaluation and T cell receptor gene rearrengement with polymerase chain reaction analysis -not certainly- but may help to the diagnosis of mycosis fungoides.

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PP4-134 MELANOCYTIC LESIONS ARISING WITHIN SEBORRHEIC KERATOSES Aikaterini Zioga1, Nafsika Simou1, Sevasti Kamina1, Christina Fotika2, Anna Batistatou1, Niki J Agnantis1 1 Department of Pathology, University Hospital of Ioannina, Ioannina, Greece 2 Department of Dermatology, University Hospital of Ioannina, Ioannina, Greece Background: Seborrheic keratoses are common benign epidermal neoplasms with considerable variety of histologic features. They are rarely associated with other skin lesions, such as basal and squamous cell carcinomas and solar keratoses. The precence of co-existing melanocytic lesions is extremely rare. The aim of the present study is to examine the frequency of the association between seborrheic keratosis and melanocytic lesions and the possible origin of the latter. Method: A retrospective analysis of 500 consecutive cases of histologically diagnosed seborrheic keratoses from the Department of Pathology, University Hospital of Ioannina, Greece was carried out. In all cases immunohistochemical staining for Mart-1 (Melan-A, DAKO) was performed using the EnVision system (DAKO) and DAB as chromogen. Results: In only one of the 500 cases (0.2%) an associated, arising within, malignant melanoma was identified ( superficial spreading, radial and vertical growth phase), while no benign melanocytic lesions were noted. Non-neoplastic melanocytes were immunohistochemically identified within all the seborrheic keratoses, dispersed in variable numbers. However, their number never exceeded that of the adjacent normal epidermis. Conclusion: The frequency of melanocytic lesions arising within a seborrheic keratosis is extremely low, at least in our region. The reported case of malignant melanoma could be interpreted as a collision tumor, with the melanoma spreading in the seborrheic keratosis, or as a de novo melanoma development within the pre-existing seborrheic keratosis. Evidence supporting the latter is the intriguing presence of melanocytes amongst the squamous and basaloid cells of the seborrheic keratosis, as shown. PP4-135 SQUAMOUS CELL CARCINOMA COMPLICATING NEVUS SEBACEUS OF JADASSOHN IN A CHILD Houda Mahmoudi-Brahim, Nejib Ben Yahia, Abdelmajid Dhouibi, Leila Njim, Adnene Moussa, Nada Touil, Abdelfattah Zakhama Department of Pathology, Univesitary Hospital Center, Monastir, Tunisia Background: Sebaceus nevi are uncommon congenital skin lesions with a well recognized potential for neoplastic changes. Such change, however, is rare before puberty. Method- Results: We present a case of squamous cell carcinoma arising in a naevus sebaceous of Jadassohn (NSJ) in a 12-year-old girl. Both these events are rare. We review the literature and discuss the association of malignant tumors on a NSJ, and the need of prophylactic surgical excision of this lesion during childhood. Conclusion: this case is unusual for two reasons: first, because squamous cell carcinoma is an uncommon malignancy in NSJ; and, second, because of the patient's unusually young age. PP4-136 PROLIFERATING TRICHILEMMAL TUMOR OF THE VULVA Sami Limem, AdnèNe Moussa, Leila Njim, Nada Touil, Rym Hadhri, Nejib Ben Yahia, Abdelfatteh Zakhama Department of Pathology Fattouma Bourguiba Hospital 5000 Monastir, Tunisia BACKGROUND: Proliferating trichilemmal tumors (PTTs) are rare cutaneous neoplasms that show features of typical pilar cysts

but also show extensive epithelial proliferation, variable cytologic atypia and mitotic activity. PTT are benign lesions; however, the malignant potential of PTT is controversial, as only a small number of clinically or histologically malignant PTTs have been reported. We report an unusal case of PTT occuring on the vulva of a young woman. METHODS AND RESULTS: A 33-year-old woman presented with a gradually enlarging, painless nodule of the left labium majus. The excised tumor was well demarcated, measured 2.5 cm in greatest diameter and was covered by unremarkable skin. Based on the histopathologic findings, this tumor was diagnosed as PTT, despite the presence of moderate cytologic atypia and a florid mitotic activity. A squamous cell carcinoma was ruled out in the absence of infiltrative growth and tumoral necrosis. The patient is disease-free at last follow-up. CONCLUSION: To our knowledge, this is the third reported case of such a tumor occurring in this location. The range of the biological behaviour of this tumor is discussed. Complete surgical excision and accurate follow-up in all cases of PTT are recommended as local relapses and metastatic spread are possible and the biological behaviour of this neoplasm seems to be difficult to predict. PP4-137 EXTRAMAMMARY PAGET’S DISEASE Radulescu Doinita1, Stolnicu Simona2, Dumitriu Sorin3 1 Department of Morphopathology, University of Medicine and Pharmacy ‘Gr. T. Popa ‘ Iassy, Romania 2 Department of Morphopathology ,University of Medicine and Pharmacy Tg. Mures, Romania 3 ‘St. Mary’ Children Hospital, Iassy, Romania Extramammary Paget disease (EMP) is a scaly erythematous eruption affecting apocrine gland bearing areas of the skin. It usually occurs in the genital, axillary or perianal regions. The majority of cases represent an apocrine adenocarcinoma in situ that has a high recurrence rate and may invade the dermis and then possesses metastatic potential. Primary EMP is an apocrine adenocarcinoma in situ that most likely arises from intraepidermal cells of apocrine gland ducts. We present 4 cases of EMP that occurred in the axillary, perianal and vulvar regions. The clinical findings were suggestive of eczema or Bowen’s disease. The treatment of EMP is essentially by surgical excision with clear margins. PP4-138 BASAL AND SQUAMOUS CELL CARCINOMAS: DIFFERENCES IN THE VASCULATURE, VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF), VEGF RECEPTOR-1 (VEGFR-1) AND VEGF RECEPTOR-2 (VEGFR-2) EXPRESSION Amalia Tsiatoura, Anna Batistatou, Aikaterini Zioga, Niki J Agnantis, Dimitrios Stefanou Department of Pathology, University of Ioannina, Medical School, Ioannina, Greece BACKGROUND: Basal (BCC) and squamous cell (SCC) carcinomas are the most common skin carcinomas. It is well known that there is a significant difference in the behavior of these tumor types, with SCCs but not BCCs undergoing metastatic spread. Vascular endothelial growth factor (VEGF) is a homodimeric disulfide-linked glycoprotein involved in angiogenesis. Tumor angiogenesis is essential for tumor growth and appears to play an important role in tumor progression and development of metastases. The aim of the present study was to investigate whether the different biological behavior of BCCs and SCCs is associated with differences in their microvasculature and the molecules involved in neoangiogenesis. METHOD: We performed a retrospective study in 107 patients, 51 with BCCs (27 male, 24 female, average age 69.5 years, average maximal diameter 1.13cm) and 56 with SCCs (37 male, 19 female, average age 74.3 years, average maximal diameter 1.25cm). We

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performed immunostaining on formalin-fixed, paraffin-embedded tissue sections using En Vision System (DAKO) and the monoclonal antibodies CD31 (Menarini Hellas), VEGF (VEGF Ab-3 JH121, Neomarkers, USA), VEGFR-1 (RP 077, DBS, USA) and VEGFR-2 (RP 076, DBS, USA). DAB was used as a chromogen. The percentage of the tumor cells that exhibited a positive immunoreactivity was determined using a ×40 objective lens. At least 1000 neoplastic cells were counted in each case and a semiquantitative method was used. RESULTS: Microvessel counts and VEGF and VEGFR-2 (but not VEGFR-1) expression differed in a statistically significant manner in the two tumor types, being more prominent in the SCCs. Furthermore, there were cases of BCCs with essentially no intratumoral blood vessels and no VEGF or VEGFR-2 expression. In SCCs, increased microvessel counts and VEGF, VEGFR-2 expression was significantly associated with low differentiation. CONCLUSION: The differences in the vasculature, VEGF and VEGFR-2 expression between SCCs and BCCs may, at least in part, account for their different biological behavior. Furthermore, SCC may represent a suitable neoplasm for antiangiogenic treatment in combination with conventional therapeutic approaches. PP4-139 A PATIENT WITH SEBACEOUS CARCINOMA, SEBACEOUS EPITHELIOMA, KERATOACANTHOMA, BASAL CELL CARCINOMA, SEBORRHEIC KERATOSIS AND MULTIPLE ACTINIC KERATOSIS WITHOUT INTERNAL MALIGNANCY (MUIR-TORRE SYNDROME): A CASE REPORT Nevra Dursun, Zuhal Gucin, Fadime Bahadir, H. Esra Pasaoglu, Kemal Behzatoglu, Nuri Osman Huten, Can Isler, Ummuhan Kiremitci The Ministry of Health stanbul Training and Research Hospital, Istanbul, Turkey Muir-Torre Sydrome represents the association of multiple sebaceous tumors, or keratoacanthoma with visceral malignancies. Cutaneous neoplasms can precede or follow a diagnosis of visceral malignancy. A 75 year old man was admitted the dermatology clinic for multiple skin lesions on his face. 14 lesions were excised.We reported one sebaceous carcinoma, one sebaceous epithelioma, one keratoacanthoma, one basal cell carcinoma, one seborrheic keratosis and nine actinic keratosis. We thought this case as a Muir-Torre syndrome and searched for internal malignancies. Colonoscopy, whole body computed tomography and prostate biopsy had made. But internal malignancy could not find. This man was worked in municipal worker for 37 years, so exposed sun light for a long time. 15 years ago and 10 years two lesions were excised, 5 years ago one more lesion was excised again. This year all of 14 lesions all appeared within 1 month. Crusted papulonodular new skin lesions are devoloped after the last exicisions. The patients with multiple sebaceous tumors and keratoacanthoma must be evaluated for the presence of visceral malingnancies. Recognition of multiple sebaseous neoplasms by pathologists and communication to clinicians of its strong association with Muir-Torre syndrome is of diagnostic importance. Our patient is followed up for skin lesions and for internal malingnancies. PP4-140 PURELY CUTANEOUS ROSAI-DORFMAN DISEASE Kenani Nesrine1, Mebazaa Amel2, Yacoubi Tahar1, Denguezli Mohamed2, Belajouza Colanda2, Nouira Rafia2 1 Department of Pathology Farhat Hached Hospital Sousse,Tunisia 2 Department of Dermatology Farhat Hached Hospital Sousse 4000, Tunisia Background: Rosai-Dorfman disease (RDD) is a rare, benign, histiocytic disorder of unknown etiology. It’s usually characterized by extensive painless lymphadenopathy often

associated with many systemic manifestations. Extranodal involvement is seen in 25 to 43% of cases. Although the skin is the most common site of extranodal involvement, purely cutaneous RDD is very rare. We report a case of a 12-year-old girl who presented a particular purely cutaneous RDD. Methods: Among a rare case of purely cutaneous RDD, we discuss the epidemiological features, clinicopathological manifestations and treatment of this disease. Results: A12-year-old girl with a 3-month history of a widespread non pruriginous papulo-nodular eruption consulted in our department of dermatology. Physical examination revealed a profusion of firm erythematous papules and nodules, distributed mainly on cheeks, perioral region, the upper trunk, thighs and legs, mimicking a molluscum contagiosum eruption. No fever or lymphadenopathy was noted. Complete blood count was normal. Erythrocyte sedimentation rate was elevated to 60 mm/hr. Serum protein electrophoresis showed polyclonal hypergammaglobulinemia. Skin biopsy showed a reticular dermal nodular infiltration by lymphocytes, neutrophils and large vacuolated histiocytes with emperipolesis. Histiocytes were positive for PS-100 and CD68, and negative for CD1a. No systemic involvement was detected within a systemic work-up. These features were consistent with the diagnosis of purely cutaneous RDD. Because of the profusion of skin lesions, the patient was treated 20mg daily with acitretine for 4 months and surgical excision of large lesions. Skin lesions cleared progressively and continued to clear spontaneously after treatment interruption. Conclusion: Cutaneous variant of RDD (CRDD) has been rarely reported. About 3% of patients have disease detectable only in the skin. CRDD affects habitually older people with a preference for white females. Clinical features of this disease are nonspecific and there are no cutaneous sites of predilection. Typical histologic features are usually present in CRDD. Immunohistochemical studies are of great interest for the diagnosis. PP4-141 HISTOPATHOLOGIC CLASSIFICATION IN BASAL CELL CARCINOMA AND COMPARISON OF P53 AND BCL-2 STAINING PATTERNS BETWEEN HISTOLOGIC GROUPS Seda Yamak, Muzeyyen Astarci, Haluk Pulat, Mehmet Gonultas, Elif Ozer, Muzaffer Caydere, Huseyin Ustun Ankara Education and Research Hospital, Ankara, Turkey Basal cell carcinoma (BCC) is a skin tumor originating from pluripotential primordial germ cells in epidermis and skin appendage. It is usually local invasive and rarely makes metastasis. However, many studies were carried out with respect to this tumor due to its characteristics of considerable morbidity, risk of developing a secondary BCC in individuals having this malignity and rare but potential risk of causing metastasis. In several studies, it was presented that aggressive BCC’s expressed more p53 and stained lesser with bcl-2. In our study, 75 patients, who were diagnosed as BCC between 2001-2005 at Pathology Department of Ankara Education and Research Hospital of the Ministry of Health were re-examined. Patterns were evaluated in respect of age, gender, localization, histological subtypes, staining patterns with p53 and bcl-2, ulcer, inflammation and presence of pigment. Although our findings do not indicate statistically satisfactory results, higher rate of staining by using bcl-2 and lower rate of staining by using p53 was noticed in non-aggressive histologic groups. However, there was no considerable difference between two markers in aggressive tumor sub-types. Our study pointed out that expression of bcl-2 and p53 may not be used as identifiers indicating aggressive or non-aggressive progress in BCC.

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PP4-142 EPIDEMIC AND CLINICO-PATHOLOGIC PROFILES OF MELANOMA IN CENTRAL TUNISIA Kenani Nesrine1, Said Sana2, Ghariani Najet2, Kebir Fatima Zahra1, Sriha Badreddine1, Belajouza Colandane2, Nouira Rafia2 1 Department of Pathology Farhat Hached Hospital Sousse,Tunisia 2 Department of Dermatology Farhat Hached Hospital Sousse 4000, Tunisia Background: Melanoma is a malignant cutaneous tumor characterised by an important metastatic potential. The aim of the present investigation was to report the epidemic and clinico-pathologic profiles of melanoma in Tunisia. Methods: A retrospective study was made recording all patients diagnosed as melanoma at the dermatology and the pathology departments of Farhat Hached hospital in Sousse, from January 1985 to December 2006. Results: Fifty nine patients were assessed with a mean age of 60,7 years (range: 5-92 years). Women were more commonly affected, the sex ratio being 1,36 (34F/25M). Consultation average was 16 months (range: 15 days- 8 years). The melanoma occurred in 66% (n=39) on a normal skin. It was localised on: extremities in 51,7 % (n=30), legs in19,1% (n=11), trunk in13,8% (n=8), face and hair in 13,8% (n=8). One case of nail’s melanoma was noted. The most common clinico-pathologic variant was the acro-lentiginous melanoma (ALM) diagnosed in 30 cases (51,7%). The other variants were: nodular melanoma (NM) in 16 cases (28%), superficial spreading melanoma (SSM) in 9 cases (16%) and lentiginous malignant melanoma (LMM) in one case (1,8%). Histo-pronostic criteria showed Clark level higher than IV in 75,6% (n=34) of patients. The mean Breslow was 7,15 mm (range: 0,6-15 mm). Conclusion: The melanoma is considered as the most malignant cutaneous tumor. InTunisia, the most frequent clinico-pathologic variant is represented by the acro-lentiginous melanoma (ALM) estimated to 51,7 % in our study, whereas the SSM (superficial spreading melanoma) is the most frequent clinical variant (70%) reported in other countries. Unfortunately, this disease is frequently diagnosed late, that’s why prognostic indicators are pejorative. Educational programs are important to be established in order to improve the early detection of this tumor. PP4-143 FOLLICULAR MUCINOSIS AND HODGKIN DISEASE Luis Alfaro1, Maria Jose Roca2, Silvia Tena1 1 Hospital Rey Don Jaime Castellon, Spain 2 Hospital Lluys Alcanys Xativa, Spain Follicular mucinosis is a peculiar cutaneous disorder, usually appearing as an incidental finding, but uncommonly associated with mycosis fungoides. Follicular mucinosis has been discovered besides in patients with malignant diseases, such as Kaposi sarcoma, chronic lymphocytic leukaemia, and Hodgkin’s disease. We had this association developed in a 38-year-old male, who presented with lymph node enlargement in cervical, axillar, and inguinal regions, and simultaneously with cutaneous lesions in face and neck. A 2 cm axillar lymph node was excised, and also a cutaneous biopsy of the lesions was taken. The skin showed a normal epidermal layer, and a superficial and medium dermal inflammatory infiltrate. Lymphocytes and some eosinophils adopted and interstitial pattern, with focal extension to follicular epithelium. There were mucin deposits inside the follicles and dissecting epithelial cells. They stained with colloidal iron. Neither neoplastic lymphocytes nor atypical cells were seen. The lymph node showed on the histological study an architectural lack of structuration, with large nodules of small and medium round cells, and only scattered original follicles, with small germinal centres, entrapped among the rest of cellularity. Some cells had big nuclei, folded and multilobated, with pale chromatin, and small nucleoli, with the typical form of LH cells of lymphocyte predominance Hodgkin’s lymphoma. Immunohistochemical study revealed B-cell markers expression

in both small and LH cells, with minimum presence of reactive T CD3-positive cells. CD30 and EMA were negative, and bcl2 had a faint expression without nodular enfacement. The diagnosis was follicular mucinosis and nodular lymphocyte predominance Hodgkin’s disease. Subsequent studies were done, and a bone marrow biopsy showed lymphomatous infiltration with nodular pattern similar to the one in lymph node. Although the association described is an uncommon event, there may have special significance based in the fact that several studies suggested a possible poor prognosis in patients with this two conditions. Our case seems to be in the same line, because the wide lymphomatous dissemination at the time of diagnosis, is not the usual presentation for lymphocyte predominance Hodgkin’s disease. PP4-144 SUPERFICIAL GRANULOMATOUS PYODERMA GANGRENOSUM M. Nieves Saracibar, Blanca Caton, Julia De Diego, Nagore Arbide Hospital Santaigo Apostol, Spain Pyoderma gangrenosum (pg) is an idiopathic neutrophilic dermatosis described by brunsting et al, in 1930. Since its description, the pathogenesis of pg has remained obscure even as an ever-widening array of systemic diseases has been described in association with it. Classic pg can occur on any skin surface, but is most commonly seen on the legs, with one or more ulcers with raised edges. genitalia may be involved. Superficial granulomatous pyoderma is a variant of pg, important to recognize and differentiate from classic, with better prognosis and not associated with underlying diseases. there are approximately 55 cases in the literature. The most important histopahological characteristic is the granulomatous pattern and his diferential diagnosis with others granulomatous dermatosis. clinical history: A 78 year old male patient with type 2 diabetes mellitus developped a nodular lesion in his penis.clinical suspicion of carcinoma. after non-conclusive biopsies, a partial penectomy was performed. later he developed more lesions in scrotum. Histologically shows granulomatous formation with sinus tracts and numerous imflamatory cells and plasma cell infiltration. The differential diagnosis included fungal or mycobarcterial infection, bromoderma, ulcerative sarcoidosis and other types of cutaneous granulomatous disorders. We have to think in pgg in cases of granulomatous pattern even in atypical localizations. PP4-145 IMMUNOHISTOCHEMISTRY HELPS DIFFERENTIATING HALO NEVUS FROM MELANOMAS WITH BRISK TUMOR INFILTRATING LYMPHOCYTES Ozlem Erdem1, Victor G Prieto2 1 Gazi University Faculty of Medicine Department of Pathology, Ankara, Turkey 2 University of Texas M. D. Anderson Cancer Centre Department of Pathology, USA Background: Lymphocytic infiltration that accompanies melanocytic lesions causes difficulty in distinction between malignant and benign lesions on conventional histopathology. Although labeling of HMB-45 and expression of Ki-67 in benign and malignant melanocytic lesions has been investigated in many studies, little is known about the expression of these markers in melanocytic lesion with dense lymphocytic infiltrate. In this study; we aimed to investigate whether there is a possible differential expression of HMB-45 and Ki-67 in benign and malignant melanocytic lesions with or without intense lymphocytic infiltration. Methods: Immunohistochemistry was performed using formalin-fixed, paraffin-embedded tissue blocks from 68 melanocytic lesions; 13 halo nevi, 7 dysplastic nevi with

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halo phenomenon, 17 dysplastic nevi without halo phenomenon, 15 invasive malignant melanoma with brisk lymphocytic infiltrate and 14 invasive malignant melanoma with nonbrisk lymphocytic infiltrate. Results: Both halo nevus and dysplastic nevus cases showed the same HMB-45 labeling pattern; strong labeling in the junctional component but only focal or weak in superficial dermal component. In melanomas, however, HMB-45 labeling was also evident in deep dermis as well superficial dermis. The number of Ki-67 positive cells in the melanoma cases with non-brisk infiltrate was significantly higher than that observed in any of the melanocytic lesions studied (p<0.001). Kaplan-Meier survival analysis revealed that neither Ki-67 expression nor HMB-45 labeling correlated with overall survival in melanomas. Conclusions: An immunohistochemical panel, including HMB-45 and Ki-67 might be useful in evaluating melanocytic lesions characterized by intense lymhpocytic infiltration. PP4-146 TWO CASES WITH T PSEUDOLYMPHOMA Ebru Zemheri, Tulay Zenginkinet, Ilkin Zindanci, Haydar Yalman, Mukaddes Kavala Istanbul Goztepe Training and Research Hospital, Istanbul, Turkey Background: Cutaneous pseudolymphomas encompass a heterogeneous group of benign reactive lymphoproliferative diseases of T- and B-cell which is clinically and histopathologically resembling malignant lymphomas. The clinical appearance of pseudolymphoma is variable and usually characterized by single or multiple, and small or large red-purple doughy or firm bumps that are usually located on the head and neck, but it can involve any area of the body. Histopathologic apperance of T-cell pseudolymphomas are characterized by a superficial bandlike infiltrate or, rarely, by a nodular pattern located in the upper dermis. Cases: 18 year-old patient with erythematous, indurated nodules on the right appearance of chin and 35 year-old patient with hyperpigmented dome shaped nodul on the nose were taken biopsy. Histologically and immunologically, we detected T-cell pseudolymphoma in both cases. Conclusion: Classification of cutaneous lymphoproliferative disorders can only be achieved by a combination of clinical, histopathologic, immunophenotypic investigation. The purpose of this article is to summarize the criteria in the differential diagnosis of benign or malignant T-cell lymphoid infiltrates of the skin. We reported two patients who had been histopathologically diagnosed as T-cell pseudolymphoma. Key words: T-cell, pseudolymphoma, skin PP4-147 EPIDERMAL LANGERHANS CELLS AND DERMAL MAST CELLS IN BENIGN AND MALIGNANT NEOPLASTIC SKIN LESIONS Canten Tataroglu1, Nil Culhaci1, Emel Dikicioglu Cetin1, Zehra Kesen2, Hicran Turhan1 1 Adnan Menderes University Medical Faculty, Aydın, Turkey 2 Denizli State Hospital, Denizli, Turkey Immune cells of the epidermis and dermis participate in the tumor progression and development. Langerhans cells are epidermal antigen presenting dendritic cells. Mast cells represent a critical role in the immune system in the dermis. In the present study, we aimed to evaluate mast cells and Langerhans cells in some skin lesions including seborrheic keratosis, keratoacanthoma, actinic keratosis and squamous cell carcinoma. This study included 12 patients with seborrheic keratosis, 9 patients with actinic keratosis , 15 patients with keratoacanthoma and 35 patients with squamous cell carcinoma. Dermal mast cells and epidermal Langerhans cells were labeled with mast cell tryptase and S100 antibodies. In the data analysis, mast cells and Langerhans cells were counted and averaged in three

representative area at x200 magnification. The number of the mast cells was significantly increased in the patients with squamous cell carcinoma in comparison with seborrheic keratosis (p:0.001). However, this parameter did not show significant difference between patients with seborrheic keratosis and actinic keratosis and between patients with seborrheic keratosis and keratoacanthoma (p=0.733, p=0.417). The number of the mast cells obtained from patients with squamous cell carcinoma did not show significant difference among patients with actinic keratosis and keratoacanthoma (p=0.986, p=0.079). The number of the Langerhans cells obtained from the patients with squamous cell carcinoma was significantly increased in comparison with seborrheic keratosis (p=0.023). Any difference was not observed between the patients with actinic keratosis and keratoacanthoma (p=0.731, p=0.601). Nevertheless, the number of the Langerhans cells was increased in the patients with squamous cell carcinoma in comparison with actinic keratosis (p=0.002). The numbers of the Langerhans cells and mast cells showed a lineer correlation only in the patients with actinic keratosis (r:0.8, p:0.008). Our data suggested that both epidermal Langerhans cells and dermal mast cells which have some contributions in the immune system, may have role in progression, development or regression of some skin tumors. PP4-148 THE STAINING METHODS OF CHITOSAN AS A NATURAL BIOPOLYMER Emine Salva1, Naziye Ozkan2, Fulya Cakalagaoglu2, Berna Karakoyun3, Julide Akbuga4 1 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department, Haydarpasa, stanbul-Turkey; Marmara University, Department of

Pharmaceutical Biotechnology, Faculty of Pharmacy, Haydarpasa, stanbul, Turkey 2 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department, Haydarpasa, stanbul, Turkey

3 Marmara University, Nursing School, Haydarpasa, stanbul-Turkey 4 Marmara University, Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Haydarpasa, stanbul, Turkey. Background: Chitosan is a natural cationic linear polymer that has recently emerged as an alternative nonviral gene delivery system.It has been advantageous biological properties in the application as wound dressing, namely biocompatibiltiy, biodegradability, anti-infectional activity and a property to accelarate wound healing.In experimental animal models, chitosan was shown to influence all stages of wound repair.The hemostatic activity of chitosan can be seen in the inflammatory phase. It also interacts with and regulates the migration of neutrophils and macrophages acting on repairing process. During the inflammatory stage, chitosan accelerates the infiltration of inflammatory cells.At the new tissue formation period, formation of granulation tissue takes place within the wound space. We have researched the different methods for staining chitosan in the experimental skin wound model.Histological analyses that performed for assessment in wound healing of chitosan products are important.Therefore, it is necessity a simple and effective method for proper histological identification of chitosan materials. Method: For skin topical application, Wistar rats were first anesthetized, their backs shaved and full thickness skin wound was made at two individual sites on the dorsum of the back of each animal.Thereafter, 100μg LMW chitosan per wound sites were topically given.Skin samples were harvested 24,48,72hours (acute term) and 1,2weeks (chronic term) after application.Specimens were paraffin embedded, sectioned and able to observe specific staining of the chitosan particles with Cibacron-Brillant Red-3BA and Iron Hematoxylin and also Safranin-O/fast Green and Iron Hematoxylin staining methods.

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Results: CBR-3BA iron hematoxylin was used for identifying the presence of chitosan incorporated with chitosan-given to rats.Chitosan was observed as red particles, iron hematoxylin was resulted blue coloration of nuclei.Cytoplasm and collagen fibrils were appeared pink color.The second method for specific staining of chitosan used the Safranin-O/Fast Green.Chitosan particles was appeared green color and with iron hematoxylin yielded stained nuclei. Conclusion. Chitosan that is an attractive material in biotechnology, used widespread fields as tissue engineering and drug delivery.Therefore, simple and specific staining methods are critical.This staining methods were highly successful in locating small quantities of chitosan in a dermal wound model.These methods may key a role progressing in the histological study of chitosan-based biotechnological products. PP4-149 BCL-2 OVEREXPRESSION AT LYMPHOCYTES IN SKIN LESION AND PERIPHERAL BLOOD IN PATIENT WITH PSORIASIS VULGARIS Nebil Bal1, lhan Tuncer2, Mete Baba3, Filiz Bolat2 1 Baskent University Medical Faculty Department of Pathology Ankara, Turkey 2 Baskent University Medical Faculty Department of Pathology Ankara, Turkey 3 Baskent University Medical Faculty Department of Dermatology Ankara, Turkey Psoriasis is a chronic inflammatory skin disease which is characterized with healing and relapsing processes. Cellular immunity mediated by T cells is accused in pathogenesis. Bcl-2 is an anti-apoptotic gene related to the bcl-2 gene family, and its overexpression in lymphocytes has been shown in some chronic inflammatory and neoplastic diseases. 39 patient with psoriasis vulgaris were taken for study group and bcl-2 expression was investigated in skin biopsies by immunohistochemically which was taken from the active skin lesion and in peripheral blood by flowcytometric analysis. It was planned that to be done flowcytometric analysis before and after the topical steroid treatment. 10 pilonidal sinus cases were selected as negative control group for immunohistochemistry and 10 patient with active Systemic Lupus erythematosus (SLE) as positive control group for flowcytometric analysis. Bcl-2 overexpression was determined in dermal lymphocytes in different intensities in all the psoriatic skin lesions except one but not found in control groups. Flowcytometric analyses could be performed in 18 patients with psoriasis vulgaris before the treatment and in 10 of 18 patients after the treatment. Mean bcl-2 expression value was found 86.82 % at the beginig and 86.02 % after the treatment and 89,3 % in groups of active SLE patient. Significant correlation was found between psoriasis vulgaris and active SLE groups. The presence high levels of bcl-2 overexpression in dermal lymphocytes and in peripheral blood which was taken before the treatment thought that the bcl-2 overexpression may have an important role in the pathogenesis of psoriasis vulgaris. Patient with active lesion of psoriasis vulgaris exhibited levels of bcl-2 expression similar to those in active SLE, suggesting that raised bcl-2 levels may be characteristics of the inflammatory diseases. Although the skin lesions have improved with topical steroid therapy, the continuity of high levels of bcl-2 expression in peripheral blood indicate the insufficiency of the topical treatment. In conclusion, we speculated that systemic steroid therapy may be useful to reduce the bcl-2 expressing T lymphocytes levels in peripheral blood and that alternative theapy may be more effective in the treatment or in prevention of recurrences and systemic complications of psoriasis vulgaris. Further studies about the effects of systemic treatment on bcl-2 expression will be helpful.

PP4-150 EXTRAOCULAR SEBACEOUS CARCINOMA ARISING FROM THE SCALP Chaabouni Najla1, Sassi Samia1, Abbes Imen1, Dhouib Rym1, Driss Maha1, Mrad Karima1, Labben Naceur2, Ben Romdhane Khaled1 1 Department of Pathology, Salah Azaeiz Institute, Bab Saadoun, Tunis, Tunisia 2 Department of Pathology,Habib Thameur Hospital,Tunis,Tunisia Sebaceous carcinomas of the skin are rare tumors. Most of them often occur on the skin of the head and the neck. They are slow-growing, locally aggressive and capable of metastatic spread. They exhibit a variety of histologic growth patterns which may cause differential diagnosis hardship. We describe a case of grade II sebaceous carcinoma in a 67-year-old Tunisian woman localized on the scalp. It was associated with a metaplastic squamous component that has led initially to a diagnosis of degenerated trichilemmal cyst or squamous cell carcinoma. But, histochemical (ORO +: oil red O stain for lipid) and immunohistochemical (EMA+, ACE -, CD15-) studies using formalin-fixed and paraffin-embedded tissue specimens confirmed the the diagnosis of sebaceous carcinoma. We discuss clinical, histological, histochemical and immunohistochemical profiles of this aggressive neoplasm. PP4-151 HAEMORRHOIDS AND GRANULOMA INGUINALE Stephanos Milias1, Epaminondas Molyvas1, Constantinos Milias2, Kyparrisis Batsios3 1 Department of Pathology 424 General Military Hospital, Thessaloniki, Greece 2 2nd Surgical Department 424 General Military Hospital, Thessaloniki, Greece 3 Department of Dermatology, 424 General Military Hospital, Thessaloniki, Greece Background: To present an unusal case of granuloma inguinale presented on the surface of haemorrhoids. Method: The patient was male, 46 years old, presented to the surgical department of our hospital, due to enlarging of haemorroids. The physical examination confirmed that there was cirsoid ectasia of the haemorroid veins. There was also an extended skin rash of the perigenital and perianal area. The patient underwent haemorrhoidectomy. Results: Four reddish nodules, 2.2-3.4 cm in greatest diameter were received. On the surface small, whitish, elevated lesions were observed. Microscopic examination revealed the usual findings of the veins of the submucosal. The mucosa was infiltrated with many lymphocytes, plasma cells and neutrophills, sometimes in small aggregates. There were also many histiocytes. The squamous epithelium showed pseudoepitheliomatous hyperplasia and focal small ulcerations. The Giemsa stain revealed that the histiocytes contained small intracytoplasmatic inclusion bodies which considered to be Donovan bodies. The patient was then examined by a Dermatologist who agreed with the histological diagnosis and added Doxycyclin in the treatment. Conclusion: Donovanosis is a tropical endemic disease, rare in the West countries. It is a sexually transmitted disease caused by Calymmatobacterium granulomatis. It usually affects the perianal and perigenital area. To our knowledge is not usual on the surface of haemorrhoids.

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PP4-152 A CASE OF MONOCYTIC LEUKAEMIA CUTIS IN A PATIENT WITH MYELODYPLASIC SYNDROME TRANSFORMING TO ACUTE MYELOID LEUKAEMIA Zribi Jihène1, Ayadi Lobna2, Makni Saloua2, Bellaaj Hatem3, Kallel Rim4, Khabir Abdelmajid4, Gouiaa Nourez4, Sellami-Boudawara Tahya4 1 Pathology Department Habib Bourguiba Hospital, Tunisia 2 Pathlogy Department Habib Bourguiba Hospital, Tunisia 3 Hematology Department Hedy Chaker Hospital, Tunisia 4 Pathology Department Habib Bourguiba Hospital, Tunisia Background: Leukaemias are a group of disorders characterized by the presence of white blood neoplasic cells in the blood stream and in bone marrow. Rarely infiltration of skin may be observed. Occasionally, skin lesions may precede haematological manifestations or may be concomitant with the diagnosis of systemic leukaemia; we report a new case of monocytic leukaeia cutis in a patient with myelodysplasic syndrome. Case report: A 66-year-old- women with a 3-year- history of myelodysplasic syndrome with abnormal bone marrow cytogenetics. Over the previous few months she had developed cutaneous lesions on the buttok and thigh; the skin biopsy showed a diffuse infiltrate of cells with bean-shaped nuclei in the upper dermis extending to the subcutis. Immunohistochemistry showed positivity for CD15, CD68 and CD 34. Immunostainingg for CD3 and CD20 was negative. These results were consistent with leukaemia cutis. Conclusion: Leukaemia cutis as the initial clinical presentation of a transforming myelodysplastic syndrome into acute myeloid leukaemia has been reported only very rarely. Our case underlines the need to look meticulously for skin changes and perform early skin biopsies in haematological patients. Skin lesions can represent the first clinical signs of an otherwise not evident bone marrow disorder. PP4-153 BETA-CATENIN AND CYCLIN D1 EXPRESSION IN THE DETERMINATION OF THE BIOLOGICAL BEHAVIOUR OF KERATOACANTHOMA Nese Calli-Demirkan1, Aysegul Aksoy-Altinboga1, Metin Akbulut1, Ferda Bir1, Nilay Sen-Turk1, Nagihan Yalcin1, Inci Gokalan-Kara2 1 Department of Pathology, School of Medicine, University of Pamukkale, Denizli, Turkey 2 Department of Plastic and Reconstructive Surgery, School of Medicine, University of Pamukkale, Denizli, Turkey Background: Keratoacanthoma (KA) is a benign cutaneous tumor that has rapid growth phase similar to the malignant tumors. KA and squamous cell carcinoma (SCC) are sometimes difficult to distinguish by histopathological examination. The purpose of this study is to determine some histopathological criteria to make a distinction between SCC and KA and the pattern of -catenin and cyclin D1 immunostaining to evaluate its potential value in difficult cases Material and methods: Immunoperoxidase staining of -catenin, and cyclin D1 were performed at paraffin-embedded sections of 14 KAs taken from archival material. On reviewing the histology, seven of the 14 KAs were characterized as KA, and the rest as KA resembling SCC. Additionally, 16 well differentiated superficial spreading SCCs were examined. We evaluated macroscopical tumor characteristics (duration of the lesion, diameter, ulceration, symmetricity, central keratin crater) and microscopical findings at each group. Results: Ulceration was found more frequent in SCC(p=0,03) and the symetricity in KAs (p=0,005). Central keratin crater, duration of the lesion or intraepithelial polymorphonuclear abscesses were not significant parameters in the distinction between KA and SCC. Immunohistochemical analysis showed that; in KA group, 6/7 (%85,7) , in KA resembling SCC 1/6 SCCs (%16,7) and in SCC group 5/16 (%31,3) were membranous -catenin positive in the deeper parts of the lesion . So; when the lesion became invasive,

the deeper part of the lesion lost membranous localization of -catenin(p=0,02). Cyclin-D1 expression was not significant between any groups. Conclusion: These findings suggest that macroscopical findings such as ulceration and symetricity of the lesion are more important than the central keratin crater, intraepithelial polymorphonuclear abscesses or duration of the lesion so as to distinguish KA from SCC. Additionally the loss of membraneous -catenin expression at the deeper part of lesion may be helpful at the differential diagnosis between KA and KA resembling SCC. PP4-154 CORRELATION OF EXPRESSION OF GROWTH HORMONE RECEPTOR AND GROWTH HORMONE ON DYSPLASTIC NAEVUS AND SKIN MALIGNANT MELANOMA CELLS Danko Mueller1, Cedna Tomasovic-Loncaric1, Bozo Kruslin2 1 Department of Pathology, University Hospital Dubrava, Zagreb, Croatia 2 Institute of Pathology “Ljudevit Jurak”, University Hospital “Sisters of Mercy”, Zagreb, Croatia AIM OF THE STUDY: To investigate possible differences in expression of growth hormone and its receptor among normal skin, dysplastic naevus and malignant melanoma group. MATERIAL AND METHODS: Twenty cases, ten males and ten females, from each of above mentioned groups were examined to analyse expression of growth hormone and its receptor. The results of immunohistochemistry were shown in semiquantitative manner and were analysed with help of statistical methods, including x2 – test and Kruskal-Wallis ANOVA test, with the level of importance <0,05. RESULTS AND CONCLUSION: The results showed that there was a higher expression of growth hormone receptor in malignant melanoma material, compared with dysplastic naevus and normal skin. Less impressive were results with growth hormone, although very similar pattern of immunohistochemical expression has been detected among mentioned groups. This was confirmed with both x2 – test and Kruskal-Wallis ANOVA test. Therefore we conclude that there might be a possibility to intervene the course of malignant melanoma and dysplastic naevus via growth hormone receptor antagonists. We strongly suggest studies on larger number of patients to be made. PP4-156 RESEARCH OF THE ANTI-SCAR ACTIVITY OF THE PREPARATION ISONIDEZUM AS A REMEDY, INHIBITING THE CONNECTIVE TISSUE FORMATION Vladimir Remish1, Elena Raevskaya2, Lina Remish3, Vasile Anestiade1 1 Scientific Centre of Pathobiology and Pathology of the Academy of Sciences of Republic Moldova, Moldova 2 Medical State University of Republic Moldova, Moldova 3 Academy of Sciences of Republic Moldova, Moldova The Preparation Isonidezum as a remedy for the preventive maintenance of the postoperative intraperetoneal adhesions is produced by the pharmaceutical enterprise “Farmako” (Moldova). We used Isonidezum in cosmetology surgery for aesthetic correction of the postoperative scars in comparison with the ointment Dermaxinab, containing 0,05% Clobetanzoli propionatum. The purpose of research - To use the preparation Isonidezum with a new purpose- for a primary preventive maintenance of postoperative scars which were considered as an non- aesthetic complication after removal the nevi. Material and methods of research. We carried out the clinical test for two groups of patients, each of which consisted of 50 individuals requiring the removal of skin benign formations. Their age was of 15-60 years and 11-55 years, in two groups respectively. For confirmation of the clinical diagnosis all the skin formations have been subjected to the morphological analysis in the respective

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groups of research: - nevocellular nevi - 34 cases (68%); seborrhea keratosis - 8 cases (16%); papillomas - 3 cases (6%); angiomas - 2 cases (4%); keloids - 2 cases (4%); dermatofibroma - 1 case (2%). In group of comparison: - nevocellular nevi - 45 cases (90%); seborrhea keratosis - 4 cases (8%); keratoacanthoma - 1 case (2%). Results of research. In the patients of the experimental group surgical treatment was accompanied by the subsequent processing of a zone of surgical intervention by the preparation Isonidezum. The borders of the wound were sutured by in layers, with the subsequent application on the wound of a sterile tampon impregnated by solution of Isonidezum. The keeping of the tampon on the wound under light pressure within 2 hours after removal of the formations was recommended. Patients of control group underwent to a usual cosmetic operation without subsequent application Isonidezum. For definition of clinical aspects postoperative scar (color, consistence, smoothness of the surface) patients of both groups were surveyed in 3 and 6 months after surgical intervention. As a result of the researches the occurrence keloids: in research group was one case, in the control group was 6 cases (p<0,05). Hence, the application of Isonidezum with these purposes showed the maximum preventive effect, no cases of the repeated referrals of the patients to doctors and cosmeticians. The remote consequences of the application of the preparation Isonidezum in these pathological conditions were not observed. PP4-157 EXPRESSION OF p14, p16 AND p53 IN RELATION TO HPV IN (PRE)MALIGNANT SQUAMOUS SKIN TUMORS Kusters-Vandevelde Heidi1, Melchers Willem1, De Koning Maurits2, Quint Wim2, De Wilde Peter1, De Jong Elke1, Aalders Sabine1, Van De Kerkhof Peter1, Blokx Willeke1 1 Radboud University Nijmegen Medical Center, The Netherlands 2 DDL Diagnostic Laboratories, Voorburg, The Netherlands Background: The tumor suppressor p14 is thought to play a key role in cell cycle control, since p14 provides a crosstalk between the two main pathways governing cell growth, namely the p14-MDM2-p53 and p16-CDK4/6-RB pathway. The expression of p14 in keratinocytic intraepidermal neoplasia (KIN) lesions has not been studied yet. Immunohistochemical studies in cervical dysplasia have reported over-expression of both p14 and p16 in high risk HPV-associated cases. Relations between HPV (human papilloma virus) and the tumor suppressors p14/p16 and p53 in skin (pre)cancer have not been clarified yet. Aims: To study 1) p14 expression, in relation with p16 and p53 expression, in (pre)malignant squamous skin tumors, and possible relations with risk factors for cutaneous carcinogenesis (sun exposure and immune status), and 2) a possible role of HPV in (pre)malignant squamous skin tumors. Materials and methods: Immunostaining for p14, p16 and p53 was performed on paraffin embedded sections of 22 low grade KIN (LKIN) lesions, 49 high grade KIN (HKIN) lesions, and 34 CSCCs from 52 renal transplant recipients (RTRs) and 53 immunocompetent individuals (ICIs). HPV detection was performed using a short PCR fragment (SPF-LiPA) assay, allowing for the detection and genotyping of 25 mucosal HPV genotypes. Beta papillomavirus detection and genotyping was carried out with the PM-PCR RHA method for identification of the 25 established beta-PV types. Results: P14 expression proved independent of the expression of both p53 and p16, irrespective of immune status and sun exposure. We found a significant association between the presence of beta-PV and sun exposure in the RTRs (p=0.002). There was no significant association between the overall presence of beta-PV and protein expression, in both the RTRs and ICIs. Conclusions: Relations between p14/p16/p53 expression and HPV presence in skin neoplasia, is different from findings in cervical neoplasia. Main causes are probably that in skin cancer, besides HPV, also sun exposure is implicated, and that in skin lesions, HPV integration in the genome is rare, in contrast to cervical cancer. Our data

support the hypothesis that HPV and ultraviolet radiation play a synergetic role in promoting carcinogenesis, probably enhanced by immunosuppression, since we found a significant association between the presence of HPV and sun exposure in the RTRs. PP4-158 PAPULAR MUCINOSIS: REPORT OF TWO CASES AND REVIEW OF THE LITERATURE Ozgur Mete1, Nesimi Buyukbabani1, Goncagul Babuna2, Can Baykal2 1 Istanbul University, Istanbul Medical Faculty, Department of Pathology, Istanbul, Turkey 2 Istanbul University, Istanbul Medical Faculty, Department of Dermatology, Istanbul, Turkey BACKGROUND: Papular mucinosis (PM) is a rare disease characterized by a symmetric distribution of erythematous to yellowish papules and/or plaques, most commonly involving the face, neck, upper trunk and arms. Some cases are associated with systemic diseases such as paraproteinaemia, dermatomyositis, scleroderma, systemic lupus erythematosus, non-Hodgkin’s lymphoma and hypothyroidism. We report herein two cases of PM. The first case was an example of PM in conjunction with Hashimoto’s thyroiditis which regressed following thyroid hormone supplementation. The second case was an example to the idiopathic form, which was not associated with any other pathology. REPORT OF THE CASES: Case 1: A 48-year-old woman had erythematous to yellowish, symmetric, 2-5 mm papules and nodules on her forearms since two months. Clinically disseminated granuloma annulare or sarcoidosis were suspected and a punch biopsy was done. Histopathological examination revealed mucin deposition in the dermis. Clinical findings were reassessed and the patient was diagnosed as a dermal mucinosis compatible with PM. Routine blood tests were unremarkable except the level of serum-free T4 10.9 pmol/L (normal range: 11-25), TSH 16.8 mIU/L (normal range: 0.4-4) and anti-thyroglobulin antibodies 379 IU/ml (normal range: 0-60 U/ml). Laboratory findings revealed hypothyroidism associated with Hashimoto’s thyroiditis. Lesions regressed frankly after nine months of thyroid hormone supplementation. Case 2: A 13-year-old girl had multiple yellowish and white, symmetric, pruritic, papular lesions on the dorsal part of fingers and hands, forearms, neck, face, and suprapubic region. Clinical diagnosis was PM. Two punch biopsies, from the dorsum of the hand and from the neck were taken. Histopathological examination revealed mucin deposition in the dermis. In the biopsy taken from the hand a fibroblastic proliferation in deep dermis accompanied mucin deposition, which was reminiscent of the scleromyxedema variant. Routine blood tests were unremarkable and paraproteinaemia was not found. Lesions regressed frankly following four months of methylprednisolone (16 mg/day) and isoretinoin (40 mg/day) therapy. CONCLUSION: It is not clear why mucin production increases in pathological conditions. Mucin is secreted in small amounts by fibroblasts in the dermis. The cause is probably multifactorial: cytokines, immunoglobulins and serum factors induce fibroblast proliferation and mucin production. It is important to remember that the diagnosis of mucinoses depends upon clinical features and biochemical investigations. PP4-159 THE EXPRESSION OF P63 AND P53 IN INTRAEPIDERMAL AND INVASIVE NEOPLASMS OF THE SKIN Damlanur Sakiz1, Tugba Taskin Turkmenoglu2, Fevziye Kabukcuoglu1 1 Department of Pathology Sisli Etfal Education and Research Hospital, Istanbul, Turkey 2 Department of Pathology Mardin State Hospital, Mardin,Turkey

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Aim: p53 is a well-known tumor supressor gene and its mutation is a common event in intraepidermal and invasive neoplasms of the skin. p63 is a homologue of the tumor supressor gene p53, which is expressed in human basal squamous epithelium. p63 plays a role in the development of squamous epithelium and, despite its homology to p53, it is considered to act as an oncogene. We evaluated p63 expression in usual skin cancers and intraepidermal neoplasms including Bowen's Disease, actinic keratosis (AK), in situ malignant melanoma (MM) an Paget's Disease to clarify the putative role of p63 expression in the development and differantial diagnosis of these lesions. Material Metod: The pathology database of the Sisli Etfal Education and Research Hospital was retrospectively reviewed to identify cases between 1999-2005. There were 17 squamous cell carcinomas (SCC), 23 basal cell carcinomas (BCC), 16 keratoacanthomas, 26 AKs, 22 Bowen's Diseases, 7 in situ MMs and 3 Paget's Diseases. In addiation, nonsunexposed normal skin tissues resected due to benign lesions acted as controls. Immunohistochemistry using antibodies against p63 and p53 was performed.Statistical analysis was done using the Kruskal Wallis, Mann Whitney and Wilcoxan Rank tests. The Spearmen test used for correlation analysis and p<.05 was considered significant. Results: There was a significant p63 staining in Bowen's Disease, AK, BCC, SCC and keratoachantomas respectively. In contrast, none of the in situ MM and Paget's Disease was positive for p63. Conclusion: Based on our findings, analysis of p63 expression may help in the differential diagnosis of Bowen's Disease and AK vs in situ MM and Paget's Disease, especially in small biopsies. PP4-160 DIAGNOSIS AND FOLLOW UP OF KERATOACANTHOMA-LIKE LESIONS: A CLINICAL-HISTOLOGIC STUDY OF 43 CASES Rf Magalhães1, Gt Cruvinel2, Gf Cintra2, Maria L. Cintra2, Appb Ismael1, Am Moraes1 1 Discipline of Dermatology, Department of Medical Clinic and School of Medical Sciences, UNICAMP, São Paulo, Brazil 2 Discipline of Dermatology, Department of Pathology, School of Medical Sciences, UNICAMP, São Paulo, Brazil Background: Keratoacanthoma (KA) is easily confused with Squamous Cell Carcinoma (SCC) either, on clinical- or histopathologic bases. However, KA undergoes spontaneous regression while SCC does not. In this way, to improve the histopathologic features that better discriminate the two tumors, clinical regression could be used as gold standard for KA. Our objective was to study the histopathologic features associated to clinical regression in KA-like lesions in which clinical course was assessed. Method: 43 biopsies of KA-like lesions were taken at patient admission. One month later, surgical excision was undergone in 18 growing lesions. Involuting lesions were left untreated even when histopathological reports of SCC were rendered. Classical histopathologic features and diagnosis were blindly recorded in both, biopsies- and surgical specimens. Results: Most lesions occured in elderly. Their settings were sun-exposed skin of head/neck and limbs. All tumors were described as having acute onset and rapid growth. No patient had recurrence following natural regression or excision. On clinical and histological basis, 32 lesions were assessed as KA and 11 as SCC. Features that indicated malignancy were observed in both groups (medium 1.63 in KAs and 6.18 in SCCs). Three out 32 KAs were assessed as KA-like undoubted SCC on blind histologic examination. No KA presented deep tumor extension beyond the level of the sweat glands. One KA showed perineural extension. Regressing stage KAs displayed less histologic signs indicative of malignancy. Conclusion: SCCs and KAs have more pathological similarities than differences. The differentiation of a KA in the proliferative stage from SCC may be impossible. The combination of the most useful features did not allow the nosologic diagnosis in difficult cases, but helped.

PP4-161 PHOTODAMAGE IN THE CAT SKIN: CLINICAL AND HISTOMORPHOMETRIC ANALYSIS Ellen M P Almeida1, Rosa A Caraca2, Geórgia F Cintra2, Randall L Adam5, Elemir M Souza3, Konradin Metze2, Maria L. Cintra2 1 Veterinary Division, Medical Sciences School, State University of Campinas (UNICAMP), São Paulo, Brazil. 2 Departments of Pathology, Medical Sciences School, State University of Campinas (UNICAMP), São Paulo, Brazil. 3 Departments of Dermatology, Medical Sciences School, State University of Campinas (UNICAMP), São Paulo, Brazil Background: Photodamage (PD) is a term that describes the array of clinical and histologic changes caused by long-term exposure to solar radiation. In human beings, although the visible manifestations take years before become clinically evident, the underlying damage occurs from the earliest exposures. The dermatologic importance of PD lies in the considerable cosmetic and psychosocial distress it causes in the older persons and in its strong presumably etiologic relationship to skin cancer. Heavy ultraviolet (UV) exposure is also harmful to animals. On animal models, most of our knowledge in the effects of UV rays, is based on mice. Cats share the same environment as their owners, and may serve as sentinels for cancer development seen in human beings. The relatively high incidence of spontaneously occurring feline solar keratoses (SK) and the significant association between SK development and white coat make the cat a good model for photodamage research in natural populations. The purpose of the present study was to investigate clinical and histomorphometric features of cat skin under long term solar exposition. Methods: Ear skin of 34 domestic shorthair cats that were chronically exposed to sun were classified, on clinical basis, as follows: group 0: normal (n= 13); group 1: initial stage of photodamage (PD) (n= 10); group 2: advanced stage of PD (n= 11). Sex and age of the cat, as well as the color of the ear that had been submitted to biopsy were recorded. Histological sections were stained with haematoxylin-eosin and microscopic features were blindly and separately recorded by 2 examiners in 10 sequential linear high power fields (HPF). Dermal cellularity was studied through a cycloid morphometric grid positioned in ten sequential HPF. An objective assessment of the epidermal thickness and relative adnexal unit area was obtained in digitalized images, with the aid of an Axiophot photomicroscope (Carl Zeiss - KS300 system). Results: Through statistical methods, older age, white-colour ear, dermal edema and sclerosis, telangiectasis, reduced adnexal area and squamotized appearance of epidermal basal cells were significantly associated to advanced stage of photodamage. There was not dermal elastosis. The animals developed thickening of the epidermis since the initial phase. Conclusion: Our results indicate a high level of skin hypersensibility to sun rays in cats. The findings may be useful for clinical tests and in general veterinary pathology and dermatology.

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Head and Neck Pathology PP4-162 DOWNREGULATION OF CD9 PROTEIN, BUT NOT OTHER TETRASPANINS, IN ORAL SQUAMOUS CELL CARCINOMA: CORRELATION WITH POOR PROGNOSIS AND DECREASED DISEASE-FREE SURVIVAL Marcilei Eliza Cavicchioli Buim1, Silvia Vanessa Lourenco2, Claudia Malheiros Coutinho-Camillo1, André Lopes Carvalho3, Fernando Augusto Soares4 1 Department of Pathology, Hospital A. C. Camargo, São Paulo, Brazil 2 Department of General Pathology, Dental School, University of São Paulo; Laboratory of Immunopathology, Institute of Tropical Medicine; Department of Dermatology, Medical School, University of São Paulo, Brazil 3 Departament of Head and Neck Surgery, Hospital A. C. Camargo, São Paulo, Brazil 4 Department of General Pathology, Dental School, University of São Paulo; Department of Pathology, Hospital A. C. Camargo, São Paulo, Brazil Background: Squamous cell carcinoma of the oral cavity (OSCC) is a commom malignancy, characterized by a high degree of agression and metastasis to cervical lymph nodes. During the process of OSCC carcinogenesis, impaired function of metastase supressor genes appear to be crucial in the progression of cancer cells to a metastatic phenotype. Tetraspanins are proteins with functional roles in a wide array of cellular processes and have been reported to be associated with the biological behavior of solid tumors, especially with their metastatic potential. Other functions include cell adhesion, motility, differentiation and proliferation. Method: The present study investigated the expression of the tetraspanins CD9, CD37, CD63, CD81 and CD82 in OCSS using immunohistochemistry. Tissue Microarray (TMA) of 127 cases of OSCC and 10 normal tissue of oral cavity were evaluated immunomorphologically and semi-quantitatively, and results were compared to the clinical-pathological features. Multivariate Cox regression, Kaplan-Meier method and qui-square test were used for statistical analysis. Results: Expression of CD63, CD37 and CD81 was not detected in the cases studied (tumors and controls). Expression of CD9 and CD82 was observed on the membrane of basal layer cells in the controls of normal oral mucosa. CD9 expression was downregulated/ negative in 42/127 (33%) OSCC cases. Loss of CD9 expression in OSCC correlated with incidence of cervical lymph node metastasis (p=0.028). A bordeline significant relationship between CD9 downregulation and advanced disease (clinical stage T3/T4) (37.0%) (p=0.055) was verified. Significance between CD9 expression and other clinicopathological features was not statistically established. Disease-free survival rate of patients with CD9 down-regulated/negative (48.6%) was significantly lower than that in patients with CD9 positive expression (71.6%) (p=0.010). The 5-year overall survival rate of patients with CD9 downregulated/negative was 44.0% against 64.0% of CD9-positive cases (p=0.071). CD82 was downregulated/negative in 100/127 (79.0%) specimens, but no correlation was observed between CD82 expression, clinicopathological parameters, disease-free survival and 5-year overall survival (p= 0.705, p= 0.549 respectively). Conclusion: Our results suggest that downregulation of CD9 might be an indicator of poor prognosis in OSCC patients.

PP4-163 MALIGNANT PERIPHERAL NERVE SHEATH TUMOUR (MPNST) OF THE NASAL CAVITY. REPORT OF A CASE AND REVIEW OF THE LITERATURE Vassilis Samaras1, Anna Tanoglidi1, Stelios Triantos2, Maria Kefala1, Georgios Papazoglou2, Calypso Barbatis1 1 Department of Pathology, Red Cross Hospital, Athens, Greece 2 Department of Otorhinolaryngology, Red Cross Hospital, Athens, Greece BACKGROUND: Peripheral nerve sheath tumours (PNST) are soft tissue neoplasms, arising from the neural sheath of autonomic, cranial, or peripheral nerves, which are rarely encountered in the nasal cavity and paranasal sinuses with 20 well-documented cases published in the English-language literature. We report a rare case of a high-grade MPNST of the nasal cavity in a woman. METHOD: A previous healthy 39-year-old woman was admitted to our hospital with left nasal obstruction without bleeding. Magnetic resonance imaging studies showed a obstructive mass of the left nasal cavity extending to the left paranasal sinuses, approaching the ipsilateral orbit with destructive malignant features. The patient underwent a total surgical removal of the mass and multiple tissue specimens were examined with hematoxylin/eosin as well as immunohistochemically for: Vimentin, Bcl-2, a-SMA, GFAP, S-100, 1-AT, C-Kit, CD34, CD31, CD57, HHV-8, Ki67, P53. RESULTS: Nasal mucosa covered by respiratory epithelium and infiltrated by a spindle cell, vascular neoplasm growing in a fascicular pattern, mimicking kaposi’s sarcoma. The tumour was unencapsulated with indistinct borders and without necrosis. The cells had oval or elongated nuclei, with rare mitotic activity and the cytoplasm was clear or fibrillary eosinophilic. There were areas of mild cellularity and a chronic inflammatory infiltrate intra and extratumoral. Immunohistochemical staining of tumour cells showed diffuse intense positivity for Vimentin, Bcl-2, a-SMA, focal reaction for GFAP, S-100, 1-AT, C-Kit and all the other markers were negative. The Ki67 index was 5% while p53 protein was overexpressed (40%). Based on the immunophenotype a diagnosis of a high grade malignant peripheral nerve sheath tumour of the nasal cavity was made. The patient received adjuvant radiation with further surgical therapy. CONCLUSION: Approximately 45% of benign PNST occur in the head and neck region. These tumours undergo malignant transformation but MPNST can form de novo. High-grade MPNST require complete surgical resection, with wide tumour free margins, combined with adjuvant irradiation and chemotherapy. However, MPNSTs are thought to be incurable with current treatment modalities because of their metastatic potential. Our case required a differential diagnosis from a wide range of spindle cell neoplasms and particularly from leiomyosarcoma. PP4-164 MALIGNANT TRANSFORMATION OF ORAL LICHEN PLANUS Spomenka Manojlovic1, Marinka Mravak Stipetic2, Ivica Luksic3, Darko Macan3, Zoran Manojlovic4 1 Department of Pathology, University Hospital Dubrava, Zagreb, Croatia 2 Department of Oral Medicine School of Dental Medicine University of Zagreb, Croatia 3 Department of Oral and Maxillofacial Surgery, University Hospital Dubrava, Zagreb, Croatia 4 Polyclinic for Clinical Pharmacology and Toxicology Bonifarm, Zagreb, Croatia INTRODUCTION: Oral lichen planus (OLP) is a chronic inflammatory disease with low, but proved risk of transformation to oral squamous cell carcinoma (OSCC). The reported rate varies from 0,4 up to 5% of cases. Oral cancer in patients with OLP has been identified as arising on the erythematous, atrophic

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and erosive lichen-planus-affected oral mucosa. Immunohistochemical and more recently genetic studies have provided evidence of cellular and molecular events in OLP lesions that could preceede malignant transformation, suggessting that molecular assessment may become a valuable clinical tool for patient's follow-up. MATERIAL AND METHODS: We present five patients with squamous cell carcinoma of the buccal mucosa arised on the pre-existing oral lichen planus. Immunohistochemical analysis of the precursor lesions was performed by using p53 and Ki-67 antigens, and compared with the unaffected mucosa of the same patient, as well as with the matched control patient's mucosa. RESULTS AND DISCUSSION: Significantly stronger expression of Ki-67 and moderatly stronger expression of p53 protein staining was seen in the overlying epithelium in the OLP lesions preceded to SCC, in comparison to the unaffected and control mucosa. Presented cases document and alert on the propensity of oral lichen planus to undergo malignant transformation and stress the importance of regular follow-up of these patients. CONCLUSION: Clinical diagnosis of OLP has to be confirmed by histopathological analysis. As a premalignant lesion, OLP need to be assessed by use of immunohistochemical and genetic markers as well. Further research is essential for better understanding of the mechanisms and propensity of some OLP lesions to undergo malignant transformation. PP4-165 CALCIFYING EPITHELIAL ODONTOGENIC TUMOR OF THE MAXILLA - CASE REPORT Spomenka Manojlovic1, Danko Mueller1, Gorana Aralica1, Jaksa Grgurevic2 1 Department of Pathology, University Hospital Dubrava, School of Medicine, University of Zagreb, Croatia 2 Department of Oral and Maxillofacial Surgery, University Hospital Dubrava, School of Dental Medicine, University of Zagreb, Croatia INTRODUCTION: Calcifying epithelial odontogenic tumor also known as Pindborg tumor is very rare, locally invasive neoplasm, characterized by amyloid-like material that may become calcified. It accounts for less than 1% of all odontogenic tumors, without gender predilection and usually clinically presents as slow-growing, painless mass, most often envolving premolar and molar region. The mandible is affected twice as often as maxilla. Patients between 2nd and 6th decade, with a mean age of 40 years are usually affected. The vast majority of cases are intraosseous, with approximately 6% arising in extraosseous locations. CLINICAL DATA: A 36-year-old Caucasian male was admitted to our Hospital because of painless swelling, which lasted over 3 months in the region of hard palate and buccal aspect of maxilla on the right side. All radiographic findings showed a tumorous mass of the right maxillary sinus with disrupted structure of the ipsilateral upper jaw and intact orbital space. The last two loose molar teeth were extracted and the biopsy was performed which confirmed the diagnosis of calcifying epithelial odontogenic tumor. PATHOHISTOLOGY: The tumor volume was about 15 cm3 and histologically it was composed of small islands and sheets of polygonal cells with abundant eosinophilic cytoplasm, clear cell borders and noticeable intercellular bridges. Nuclei were pleomorphic, without mitotic figures. Eosinophilic, homogeneous hyaline material, within and around tumor cells, was proven to be amyloid-like, because when stained with Congo red, demonstrated the classic bright green birefringence viewed with polarized light under microscope. This amyloid-like material was often centrally calcified in the form of Liesegang concentric rings. DISCUSSION: There are around 200 cases described in the literature to the present. Long-term follow up is essential because there is a recurrence risk if the tumor was incompletely resected and in particular with the clear cell variant.

PP4-166 DEVELOPMENT OF A NEW FAST CHEMOSENSITIVITY TEST IN LARYNX CARCINOMA Daniela Fanni1, Luca Pilloni1, Pier Paolo Coni1, Giancarlo Senes1, Alberto Ravarino1, Zanino Pusceddu2, Ernesto Proto2, Gavino Faa1 1 Dipartimento di Citomorfologia, Sezione Anatomia Patologica University of Cagliari, Italy 2 Dipartimento di Scienze Chirurgiche e Trapianti d’Organo, Sezione di Otorinolaringoiatria, University of Cagliari , Italy BACKGROUND: The aim of our preliminary study was to develop an accurate chemosensitivity test for larinx carcinomas (LKs). The final goal of our study was a test which could predict the effects of cisplatin on tumour cells prior the onset of treatment in order to individualize the pharmacological strategy. METHODS: We directly exposed for a short period (30 minutes) fresh samples from 3 LKs using cisplatin (Teva L01XA01) assessing 3 different cisplatin concentrations: low (6,25 μg/ml), medium (25 μg/ml) and high (75 μg/ml). We also utilized 2 control specimens, one to be immediately formalin-fixed and one to be incubated in the medium, but lacking cisplatin. Cases interpretation was performed by histological examination of H-E stained sections. The main objectives of this study were to characterize the morphological changes induced in tumour cells and to determine effects on molecular markers involved in proliferation and/or apoptosis. RESULTS: The preliminary observation of these 3 LKs allowed us to detect the elementary lesions that could have a role in the identification of the reactivity of tumour cells toward cisplatin: extracellular and/or intracellular oedema, apoptosis and at intratumoural necrosis. Although all 3 tumours showed an identical histotype and the same site of origin, case 1 and case 3 showed a striking morphological reactivity to cisplatin, on the contrary, case 2 did not show any significant morphological features due to cisplatin exposure. What’s more cases 1 and 3 were characterized by decreasing nuclear immunoreactivity for cycline D1, p53 and PCNA. Therefore, at the molecular level, case 2 did not show any significant change in immunoreactivity for cycline D1, p53 and PCNA. CONCLUSION: Our data evidence that samples from 3 LKs may evidence different patterns to cisplatin exposure. Extracellular and/or intracellular oedema, apoptosis and necrosis were the most frequent findings observed in cisplatin-treated specimens. These observations induced us to focus our attention toward the follow up of these patients in order to compare our in “in vitro” data with the clinical response of patients, although, our findings need further validation on a larger number of neoplasias. PP4-167 NGF-BETA/PRONGF AND THEIR RECEPTORS IN NORMAL HUMAN ORAL MUCOSA Katsuhiko Hayashi1, Trond Storesund2, Olav Schreurs2, Cuong Khuu2, Camilla Husvik2, Andreas Karatsaidis2, Masashi Sugisaki1, Karl Schenck2 1 Department of Dentistry, Jikei University School of Medicine, Tokyo, Japan 2 Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway BACKGROUND: Nerve growth factor (NGF- ) and its precursor proNGF are important for differentiation and survival of neurons and dermal keratinocytes. NGF- is secreted in the saliva of mouse and man. The aim of this study was to determine whether NGF can play a role in the differentiation and wound healing of oral mucosa by investigating its effects on oral keratinocytes. METHODS: mRNA production and protein expression of NGF/proNGF and their receptors TrkA and p75 NTR were analyzed in cultured human oral mucosal keratinocytes (OMK) by Western blot analysis and reverse transcribed PCR respectively. Proliferation of OMK upon NGF-

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stimulation was measured by BrdU incorporation. The effect of NGF- on the migration of OMK was studied using a scratch assay. Expression of NGF- and its receptors in biopsies from normal oral mucosa (NOM) and oral mucosal equivalents were examined using immunological staining. RESULTS: OMK expressed mRNA for NGF- /proNGF and their receptors TrkA and p75NTR. Lysates from OMK did not contain mature NGF- but several NGF-proforms with molecular weights between 32 and 114 kDa. Culture medium from OMK contained 75 kDa proNGF. Addition of NGF- significantly enhanced proliferation of OMK cultures and in vitro scratch closure. Immunostaining of biopsies from normal oral mucosa showed the presence of proNGF in all epithelial layers. NGF staining was observed in the granular and upper spinous cell layers. TrkA immunoreactivity was detected in basal and parabasal cells, with weak to moderate staining in spinous and granular cell layers. p75NTR staining was seen in basal cell layers. CONCLUSION: NGF- /proNGF (from saliva or from the keratinocytes themselves) has mitogenic and motogenic effects on OMK and may therefore aid in healing of oral wounds. Differential expression of NGF and NGF-receptors throughout the epithelium suggests a role in epithelial differentiation. PP4-168 KARYOMETRIC ANALYSIS OF POSTIRRADIATION SIALOADENITIS Dragan Mihailovic1, Zaklina Mijovic1, Milos Kostov2, Cedo Kutlesic1, Vesna Mihailovic1 1 Institute of Pathology, University of Nis, Serbia 2 Department of Pathoanatomy, Military Hospital, Nis , Serbia Aim: The aim of this study was to estimate karyometric variables in postirradiation sialoadenitis. Material and Methods. At Institute of Pathology, University of Nis, four patients with postirradiation sialoadenitis, and twelve patients with normal parotid gland (control group) were analyzed. Nuclear area, mean and mode optical density (OD), standard deviation of OD, minimal and maximal OD, intergrated OD (IOD), nuclear perimeter and circulariry were analyzed by ImageJ software at objective x63 on routinely stained histopathological sections. Results: Histologically, atrophy of parenchyma and fibrosis were found in patients with postirradiation sialoadenitis. In postirradiation sialoadenitis, the nuclei were more irregular in contour (circularity=0.84±0.03), compared to control group (0.88±0.02)(p<0.01). Mean and maximal ODs were significantly lower in patients with postirradiation sialoadenitis (0.39±0.04, and 0.51±0.03 a.u.), compared to control group (0.44±0.04, and 0.56±0.06 a.u., respectively)(p<0.05). Conclusion: Our results suggest that radiation therapy causes irregularities of nuclear shape and loss of chromatin density. PP4-169 CRANIAL MEASUREMENTS USED IN SEX AND AGE ESTABLISHMENT Fotios Chatzinikolaou¹, Alexandra Enache², Natalia Vladica¹ 1 Department of Pathology, Theageneio, Cancer Hospital, Thessaloniki, Greece 2 Department of Forensic Medicine, University of Timisoara, Romania Aim: The purpose of the study was to interpret and apply the most efficient parameters in establishing the sex and age of cranial fragments. Materials and Methods: Two skulls had been found in two different places, isolated, without other bones and unburied. After removing the leaves and the earth, the skulls were cleaned and measured. Results: The first skull was missing a mandible and had teeth 18, 23 ,24 and 27 with 11 and 12 lost post mortem and the other teeth lost ante mortem judging by the aspect of the alveolar margin. The second skull had the mandible, but with a fractured gonion and a coronoid process. The edentation was complete, with a partial fracture of the maximale

alveolar process. The left zigomatic region presented a cominnutive fracture with the absence of some fragments of the anterior wall of the maxilar sinus. Cranial measurements highlighted some elements specifying the sex (the size of the foramen magna, of the left orbit), but also some elements unable to surely specify the sex (cranial parameter, cranial diameters, the mastoid and the aspect of the external face of the gonion). The teeth present in the first case presented decayed processes which lowered the possibility of appreciating the abrasion and the age. In the second case, for age establishment the aspects of the cranial sutures (the obliteration degree) were used. Conclusion: The sizes of certain cranial elements can be used for establishing the sex. In the absence of the teeth, age can be presumed by the degree of obliteration of the cranial sutures and the goniac angle. PP4-170 CLEAR CELLS IN ODONTOGENIC TUMORS Sabina Zurac1, Peter De Wilde2, Pieter Slootweg2 1 Department of Pathology, Colentina University Hospital, Carol Davila University of Medicine and Pharmacy, Bucharest, Romania 2 Department of Pathology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands Background: Analysis of the English medical literature reveals a different comprehension of the clear cell (CC) component in odontogenic tumors, especially when tumors bearing both ameloblastomatous features and CC component are discussed. Four types of these tumors are described in the literature: three malignant (1. tumors entirely composed of CCs, 2. admixture of islands of CCs and cords of smaller cells, 3. admixture of islands of CCs and ameloblastomatous areas) and one benign (follicular ameloblastoma with extensive replacement of the stellate reticulum by CCs). Method: We performed an analysis of the microscopic characteristics of the CC component in 14 cases of odontogenic carcinomas (6 clear cell odontogenic carcinomas (CCOCs), 6 ameloblastic carcinomas (ACs) and 2 primary intraosseous squamous cell carcinomas (PISCCs)) versus the CC component in 30 otherwise ordinary ameloblastomas. We noticed the similar appearance of the CC in the different types of odontogenic carcinomas and we compared them with that of the CC in ameloblastomas. All statistical tests were performed with SPSS version 12.0.1 for Windows (Fisher’s exact test and non-parametric Mann-Whitney-U test) with a level of statistical significance P<0.05. Results: There is definite similarity of the CCs in malignant odontogenic tumors without ectomesenchymal contribution despite the heterogeneity of the tumoral types. The cellular characteristics are statistically different in malignant versus benign odontogenic tumors. CCs with a nuclear/cellular ratio <0.5, elongated spindle-shaped nuclei, fine granular chromatin and small nucleoli are seen in benign lesions; CCs in malignant tumors are smaller with a higher nuclear/ cellular ratio, coarse granular chromatin and prominent nucleoli. (P < 0.001, except P nuclear shape = 0.03). These findings suggest that CCs represent a phenotypic cellular variant and not the hallmark of the lesion. We propose a reassignation of different types of CCOC: - CCOC, ameloblastomatous pattern (+/- squamous differentiation) - AC, CC variant; - CCOC, biphasic/monophasic pattern + squamous differentiation - PISCC, CC variant; - CCOC, biphasic/monophasic pattern, no squamous differentiation - odontogenic carcinoma not otherwise specified. Conclusion: CCOC in its current acceptation is an exceedingly rare tumor that might be reclassified as variants of different other types of odontogenic carcinoma. However, no matter if the CCOC name is retained or discarded, one should remember that benign odontogenic tumors may display CC component as well as the malignant ones.

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PP4-171 EXPRESSION OF PROGNOSTIC AND PROLIFERATION MARKERS IN SALIVARY GLAND TUMORS Rosa Maria Garcia-Martin, Ana Belen Enguita, Cristina Murillo, Esther Conde, Fernando Lopez-Rios, Claudio Ballestin ”12 de Octubre” University Hospital, Spain Aims: The aim of the present study was to investigate the relationship between different prognostic and proliferation markers in salivary gland tumors. Methods: This study was performed to examine the expression of the antibodies, cathepsin D, methalloproteinases 1 and 2 (MMP-1, MMP-2), p53, MIB-1, PCNA, and Replication protein A (RP-A) in 100 salivary gland tumors. The expression of these antibodies was examined by immunohistochemistry in 9 normal parotid tissue, and various tumors of the salivary glands (14 pleomorphic adenomas (AP), 11 polymorphous low-grade adenocarcinoma, 16 mucoepidermoid carcinomas, 14 acinic cell carcinomas, 12 adenoid cystic carcinomas, and 8 not-otherwise-specified adenocarcinomas). Immunohistochemical studies were done on paraffin embedded tissue sections. Results: Cathepsina D antibody gave strong reactions in all neoplasms, P53 and Mib-1 were negative in normal parotid glands and AP but were highly expressed in all malignant tumors. MMP-1 was negative in normal parotid glands, and we observed low expression in AP, and high expression in carcinomas. MMP-2 was negative in normal salivary glands, whereas the expression was higher in both pleomorphic adenomas and carcinomas. Between pleomorphic adenomas, the myxoid type expressed MMP-1 and MMP-2 at higher levels than the cellular ones, although this latter type were showed significant expression of RP-A. The expression of MMP-2 in myxoid pleomorphic adenomas was similar to that of carcinomas. The expression of MMP-2, P53, MIB-1 and RP-A in high grade mucoepidermoid carcinomas was higher than in the low grade ones, although in this latter type, the protein expression of MMP-1 was high. RP-A expression was low in carcinomas. This protein plays an important role in DNA replication, recombination and repair. Conclusions: 1. - Pleomorphic adenomas of the myxoid type have an expression of MMPs similar to that of carcinomas. 2. - MMP-1 expression was low or negative in normal salivary glands and in pleomorphic adenomas, but was high in carcinomas. 3. - MMP-2 expression was negative in normal salivary glands, but was higher in pleomorphic adenomas than in carcinomas. 4. - Low grade mucoepidermoid carcinomas exhibited high MMP-1 expression whereas MMP-2 expression was high in high grade mucoepidermoid carcinomas. 5,-RP- A expression was low in carcinomas. PP4-172 PROGNOSTIC SIGNIFICANCE OF FAS AND p53 IN THE SQUAMOUS CELL CARCINOMA OF THE LARYNX M. Hakan Karabulut1, Medine Murtazaoglu2, Dilek Yavuzer1, Aylin Ege Gul1, Nimet Karadayi1 1 Dr. Lutfi Kirdar Kartal Education and Research Hospital, Pathology Department, Istanbul, Turkey 2 Hakkari State Hospital, Pathology Department, Hakkari, Turkey Background; Fas receptor expression makes tumor cells susceptible to the host immune system. In some cases of the squamous cell carcinoma of the head and neck, Fas has reported to be down-regulated, but about its relationship with clinicopathologic parameters and prognostic predictive value, controversy still remains. p53 is the most commonly mutated gene in human cancer, its prognostic and diagnostic value has been studied and contradictory results reported especially in the head and neck cancers. Method; Paraffin embedded tissue blocks of 53 primary squamous cell carcinoma of the larynx were investigated for expression of Fas and p53 by immunohistochemistry. Relationship of these two proteins with the clinicopathologic data has been studied. Results; The degree of Fas expression revealed a positive relationship with the

histologic differentiation and there was a positive relationship with the expression of p53 and tumor size. We were unable to show statistically significant relationship with other prognostic parameters and these two proteins. Conclusion; Increased expression of Fas protein seems to have a positive effect on histologic differentiation and expression of p53 may have a role on the tumor progression with its possible effect on tumor size. But because of the lack of significant relationship with the majority of the prognostic parameters for both of these two markers, their prognostic value seems to be limited. PP4-173 SYNCHRONIC TUMORS OF THE LARYNX: LEIOMYOSARCOMA AND SQUAMOUS CELL CARCINOMA. A CASE REPORT Aysun Uguz1, Naciye Ozeren1, Canan Ersoz1, Fikret Cetik2 1 Cukurova University, Medical Faculty, Dept. of Pathology, Adana, Turkey 2 Cukurova University, Medical Faculty, Dept. of Otorhinolaryngology, Adana, Turkey Background: The laryngeal smooth muscle tumors are rarely seen. In the literature one case of hypopharyngeal leiomyosarcoma, and two cases of laryngeal smooth muscle tumors including one angioleiomyoma, one leiomyosarcoma and one carcinoleiomyosarcoma were reported. On the other hand, squamous cell carcinoma is the most frequent tumor of the larynx. Coexistence of mesenchymal tumor and carcinoma is extremely rare in larynx. Case Report: A case who is 51 years old man was presented. He admitted to the hospital with disphagy complaint related with his smoking history. He had been smoked two pockets of cigarettes in a day for 20 years. In the laryngoscopy a mass on the left vocal cord was observed and performed biopsy procedure. In microscopic examination, leiomyosarcoma plus squamous cell carcinoma was detected. These two kinds of tumor were been seperately and side by side but they were not observed as intermingled areas of leiomyosarcoma and squamous cell carcinoma. Immunohistochemically smooth muscle actin was positive in leiomyosarcomatous area and pansitokeratin was positive in squamous cell carcinoma area and vice versa were negative. Discussion: These tumours may present diagnostic difficulties both histopathologically and clinically. The patients usually present no characteristic symptoms in early stage and the clinical symptoms are similar to squamous cell carcinoma however immunohistochemical investigation is helpful in distinguishing smooth muscle tumours from other connective tissue neoplasms and spindle cell squamous cell carcinoma. This report describes synchronic tumors of laryngeal leiomyosarcoma and squamous cell carcinoma along with its clinical and histopathologic features. PP4-174 CORRELATION OF PCNA, Ki67 AND P53 EXPRESSIONS WITH HISTOPATHOLOGICAL GRADE IN MUCOEPIDERMOID CARCINOMA Fulya Koybasioglu1, Seyhan Ozakkoyunlu1, Evrim Ozturk2, Binnur U. Onal2, Nuray Guneri3 1 Ankara Dıskapı Training & Research Hospital, Ankara, Turkey 2 Ankara Dıskapı Training & Research Hospital, Turkey 3 Gazi University Faculty of Commerce and Tourism Education Department of Computer, Ankara, Turkey Background: Mucoepidermoid carcinoma constitutes almost 10% of all salivary gland tumours and the most common site is parotid gland. Even though the prognosis of mucoepidermoid carcinoma correlates with the histological grade, the occasional precence of metastasis in low grade tumours demonstrate that low grade histological features are not always indicative of low- grade biological behavior. In this study, PCNA, Ki67 and p53 expressions were corralated with histopathological grade of

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mucoepidermoid carcinoma. Method: We reviwed a total of 31 mucoepidermoid carcinoma cases reported in our department. The distribution of the tumours according to the glands were as follows: 19 parotid, 3 submandibular, 9 minor salivary gland. In microscopic examination, AFIP grading system yield 11 (35.5%) low grade, 11 (35.5%) intermediate and 9 (29%) high grade tumours. For statistical analysis Kruscal Wallis and Mann-Whitney U tests were used. Results: Mean Ki67 indices of histopathological grades were calculated as 1.8% for low, 2.5% for intermediate and 7% for high grade tumours. PCNA indices, on the other hand, were 36%, 43%, 66% respectively. In statistical analysis, a significant difference was obtained between the Ki67 and PCNA indices with the increasing grade of the tumor (p<0.05). P53 expressions did not demostrate any meanignful correlation with the histopathological grade. Conclusion: In conclusion, Ki67 and PCNA demonstrates a positive correlation with increasing histopathological grade in mucoepidermoid carcinoma. PP4-175 NUCLEAR PROFILES AND EXPRESSION PERCENTAGES OF P27 STAINED AND UNSTAINED TUMOR CELLS IN LARYNX CARCINOMAS: COULD IT BE USED AS A PROGNOSTIC TOOL? Guzide Ayse Gokhan1, Yildirim Karslioglu2, Caner Ozbey1, Gulay Ozbilim1, Kenan Guney3, Murat Turhan3, Kemal Hakan Gulkesen4 1 Department of Pathology, Akdeniz University School of Medicine, Antalya, Turkey 2 Department of Pathology, Gulhane Military Medical Academy, Ankara, Turkey 3 Department of Ear, Nose, Throat, Head and Neck Surgery, Akdeniz University School of Medicine, Antalya, Turkey 4 Department of Biostatistics and Medical Informatics, Akdeniz University School of Medicine, Antalya, Turkey Background: This study was carried out on a set of twenty-eight laryngeal carcinomas to explore the P27 expression levels, as well as some nuclear planimetric variables of tumor cells. Our aim was to investigate the correlation of these data with conventional histopathological and clinical prognostic factors, if any. Method: Twenty-eight (28) laryngeal squamous cell carcinomas were studied retrospectively. We applied monoclonal antibody against P27 protein using streptovidin-biotin method. In addition to the expression level of P27 in each case, some nuclear planimetric variables of immunopositive and immunonegative tumor cells were measured using a computer assisted image analysis workstation. The expression level was considered as low when the tumor had 10% of P27 immunoreactivity. The potential relationships of these data to the tumor grade, stage, survival, and lymph node metastasis were assesed statistically. Results: Seventeen cases were Grade 1 (60%), remaining 11 (40%) were Grade 2 tumors. Among them, 14 cases had lymph node metastasis. In 19 cases (61%) P27 immunoexpressions were higher than 10%. In immunopositive cells, some nuclear variables suggested lower values than those in the immunonegative ones. These were: mean area (mean: 44.18), CV (mean: 43.39), perimeter (mean:25.8), f-max (mean:9.41) and f-min (mean:6) in immunopositive cells whereas mean area (mean:53.70), cv (39.36), perimeter (28.63), f-max (mean:1.39), f-min (mean:6.65) in immunonegative cells. All of these differences were statistically significant (p<0.05). Grade 1 tumor cells’ nuclei which were labelled immunohistochemically showed significantly higher values of mean area, sd area and f-max than those in the Grade 2 ones. Differences between them are also significant (p< 0.05) Conclusion: Downregulation of P27 is known to be associated with poor prognosis in a number of malignancies. Although similar results were reported in larynx carcinomas, they are limited in number. Nuclear morphometric measurements have been reported as useful prognostic predictors in various cancers including larynx carcinomas. In this study, we

concluded that P27 immunolabeling associated with the nuclear morphometry could be useful to predict the prognosis of larynx carcinoma. Further studies in larger series are needed to make more concrete deductions. PP4-176 EXTRANODAL NK/T-CELL LYMPHOMA, NASAL TYPE Eleftheria Delliou1, Virginia Papamichail2, Thivi Vasilakaki3, Zafiroula Doudoulakaki4, Diamanto Zizi3 1 Laboratory of Pathology – Xanthi, Greece 2 Laboratory of Pathology – Piraeus, Greece 3 Department of Pathology of Tzaneio General Hospital of Piraeus, Greece 4 Department of Otolaryngology of Sismanogleio General Hospital of Komotini, Greece Objective: A case of an extranodal NK/T-cell lymphoma, nasal type is described. Methods & patients: A 75 years old female patient presented with malaise, weight loss and symptoms of nasal obstruction.. Clinical examination revealed an intranasal mass and an ulcerated lesion on hard palate with extensive mid-facial destructive lesions. Biopsy of the intranasal mass was undertaken. Results: The microscopic examination of the specimen using immunochemistry and the EBV + phenotype of the patient, suggested the presence of a T-cell lymphoproliferative lesion, characterized as NK/T-cell lymphoma, nasal type. Discussion-Conclusion: The nasal type NK/T-cell lymphoma is a rear type of predominantly extranodal non-Hodgkin lymphoma, previously known as lethal midline granuloma, which is associated with Epstein Barr virus. This lymphoma occurs most often in adults and is more common in males than females. It causes destruction of the mid-face, palatal and orbital walls. It can also involve skin, soft tissue, gastrointestinal and upper respiratory track. Histologic examination shows a diffuse lymphomatous infiltrate with angiocentric growth pattern and areas of necrosis. In most cases the lymphoma is composed of medium sized cells or a mixture of small and large cells. It must be mentioned that the cytological spectrum is very broad. Mitotic figures are easily found. Sometimes there might be a heavy admixture of inflammatory cells. The typical immunophenotype is: CD2+, CD56+ and cytoplasmic CD3+. Occasionally CD7 or CD30 are positive. CD43 and CD45RO are commonly expressed. CD4, CD5, CD57 are negative. It must be emphasized that the EBV positivity is essential for such a diagnosis. The prognosis is variable. Early stage disease may respond to radiotherapy alone, however late stage disease does not respond well to any available therapies. PP4-177 A STUDY OF THE EFFECT OF PHARMACEUTICAL SUBSTANCES ON THE NASAL MUCOSA Andreas Lazaris1, Evangelos Nikolaidis2, George Agrogiannis1, Evangelia Ferekidou3, Vasilios Papanikolaou3, Apostolos Papalois4, Sofia Tseleni1, Eleftherios Ferekidis3, Efstratios Patsouris1 1 National and Kapodistrian University of Athens, School of Medicine, 1st Dept of Pathology, Greece 2 General Hospital of Athenes “Hellenic Red Cross”, Greece 3 1st ENT Department, University of Athens, Greece 4 Experimental – Research Unit, ELPEN Pharma,Athenes,Greece Background The study of the histological changes of the nasal mucosa, caused by local administration of vasoconstrictive agents (xylometazoline), with or without nasal rinsing with enriched sea-water solutions. Method Ten PMR-Landraze pigs (half of the total study population) were used. Therapeutic doses of the studied medication were administered over the period of one month and nasal mucosa biopsies were taken in pre-set times. Hematoxylin and eosin slices were reviewed under optic microscope. Each specimen was evaluated concerning the degree of: 1.Inflammation (quota of inflammatory cell infiltration)

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2.Fibrosis (quota of slice surface covered by fibrous tissue) 3.Epithelial metaplasia (quota of squamous epithelial metaplasia) 4.Epithelial atypia (quota of atypic epithelial cells) 5. Necrosis Results Under the effect of the local acting vasocostrictive medication (xylometazoline) all of the studied parameters worsened. Similar results were observed when the administration of the agent was combined with nasal rinsing. However, inflammation worsened with a statistically significant slower rate, after ten days of combination of xylometazoline and nasal rinsing. Conclusion Preliminary results show the benefactory effect of nasal rinsing with enriched sea-water solutions, concerning the course of inflammation of nasal mucosa caused by the administration of local acting xylometazoline. PP4-178 EVALUATION OF PROLIFERATION AND APOPTOSIS RATE OF OSTEOSARCOMA OF JAW S. Elif Gultekin1, Burcu Senguven1, Omer Uluoglu2 1 Gazi University Dental Faculty Department of Oral Pathology, Turkey 2 Gazi University Medical Faculty Department of Pathology, Turkey Background: Osteosarcoma is the primary malignant bone tumour affecting more frequently long bones of young people. Approximately % 6 of all ostesorcoma occurs in the jaws. Jaw Osteosarcoma (JOS) is generally accepted to be a pathologically distinct entity due to older age at presentation, longer median survival, local reccurences and low incidence of metastasis.It has been suggested that better prognosis of JOS than long bones (LOS), may related to the type and the lower grade of the tumor. The aim of this study is to investigate the proliferation and apoptotoic rate of tumor cells in JOS by comparing LOS and discuss the possible corelation with grading, cellularity and histologic type. Methods: The study was conducted on the paraffin embedded samples of a total of 10 ostesarcoma ( 5 JOS and 5 LOS) cases. Immunohistochemistry was performed using antibody against K i67,while in situ hybridization (Tunel method) was done for detection of apoptotic cells. The number of positive cells was determined by software and histological grading was done . Results: The mean age of the patients of JOS and LOS were 26.5 and 19 years, respectively. Most of the JOS cases were from maxilla ( 80%), while femur was the most common site in LOS. 60% of JOS and 80% of LOS cases were grade 3. Proliferation rate is higher in LOS than JOS cases. Ki 67 positivity was seen 40% of JOS and 60% of LOS cases which were high grade tumors (grade 3). There was not any positivite staining for apoptosis in both tumor groups. Conclusion: Ki 67 Expression may related to the grade of JOS and have more value than apoptotic index. Studies with large series is needed to confirm these findings. PP4-179 PREVALANCE OF EBV, HPV 16 AND HSV-1 IN ORAL LICHEN PLANUS Benay Tokman, Burcu Senguven, Cem Demir Gazi University, Faculty of Dentistry, Dept. of Oral Pathology, Turkey Background: Oral lichen plnus (OLP) is a chronic inflammatory mucosal disease of unknown etiology. OLP occurs more frequently than the cutaneous form and tends to be more persistent and more resistant to treatment. Although the etiopathogenesis of lichen planus is unknown, it is generally considered to be an immunologically mediated process that histologically resembles hypersensitivity reaction. Lichen planus is probably of multifactorial origin, sometimes induced by drugs or dental materials, psyhological factors, infective agents, often idiopathic. Viral infections have recently been linked with OLP. HSV-1, CMV, HHV-6, EBV HPV, HCV are viruses that have been studied in the etiopathogenesis of OLP. The aim of the

present study was to explore a possible association of HPV16, HSV-1 and EBV with OLP. Method: The study was conducted on formalin fixed paraffin embedded tissue specimens of 65 OLP cases. Five-micrometer thick sections of formalin fixed and paraffin embedded biopsy samples were processed by the avidin-biotin-peroxidase complex (ABC) method for Herpes Simplex type 1 , Epstein Barr virus and Human Papilloma virus type 16 primary antibodies. The red-brown nuclear staining was accepted as positive staining for each antibody. To confirm HPV 16 positivity, nested PCR technique was also performed. 15 normal oral mucosal tissue specimens were used as control group. Statistical analysis were performed by Fısher’s exact test and Pearson Chi square methods. Results: The number of OLP cases that were positive for EBV was 23 ( 35%), while HPV 16 was positive in 14 (21%) cases and HSV-1 was positive in 6 (9%) cases. One case was positive for all 3 viruses. 3 cases were positve for both EBV and HSV-1, 5 cases were positive for both EBV and HPV 16, and 1 case was positive for both HSV-1 and HPV 16. The control group was totally negative for all 3 viruses. EBV and HPV 16 positivity in OLP were statistically significant. Conclusion: Although the high prevalance of EBV and HPV 16 does not imply a direct causative relation, it suggested that these viruses might be involved in the pathogenesis of OLP, while HSV positivity might be a secondary infection as a result of possible local immunosupression. EBV and HPV 16 positivity in OLP which are both oncogenic viruses should be considered seriously. PP4-180 EVALUATION OF HOST IMMUNITY BY THE DETECTION OF NATURAL KILLER CELLS AND THE EXPRESSION OF TUMOR NECROSIS FACTOR ALPHA AND ITS CORRELATION WITH PROGNOSIS IN ORAL SQUAMOUS CELL CARCINOMAS Mahiye Reyhan Turkseven, Tulin Oygur, Benay Tokman Gazi University Faculty of Dentistry, Dept. of Oral Pathology, Turkey Background: Natural Killer (NK)cells are able to show early and spontaneous cytotoxic reaction against tumor cells. NK cells have responsibility in cytokine secretion that have role in antitumor immunity. These cytokines are able to regulate NK cell activity by the otokrine effect. In addition to its well known biological effects, TNF- , also produced by NK cells, shows cytotoxic effect on some tumor cell lines. This research aimed to give a new insight to the interaction between host local immune response and the biological behaviour of the tumor by evaluation of intratumoral NK cells and TNF- expression in oral squamous cell carcinoma Method: In 46 cases of OSCC, CD57(NK) and TNF- antibodies were immunohistochemically detected. Levels of CD57 and TNF- expressions were microscopically evaluated in 2 histological groups that were categorized in accordance with their grading scores of invasive margins. Furthermore, the clinical stages of 15 cases were correlated with CD57 and TNF- expression levels. Results: While the number of CD57(+) cells was statistically lower in the group with worse prognosis, the level of TNF alpha was found statistically higher. Conclusion: Increased level of TNF alpha might have direct role on the decrease in NK cells for the high invasive group. TNF alpha that causes connective tissue destruction might support the invasion of the tumor cells. PP4-181 GIANT CELLS LESIONS - RETROSPECTIVE ANALYSIS Slobodanka Vukelic-Markovic, Zoran Mirkovic, Ruzica Kozomara, Nebojsa Jovic, Srboljub Stosic Clinic of Maxillofacial Surgery, Military Medical Academy, Belgrade, Serbia Giant cell lesions mostly originate from the soft tissues, but rarely also from the bone. Giant cell lesions of gums are peripheral

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giant cell granulomas (PGCG), but there are a number of lesions that occur in the jaws that contain giant cells within them: central giant cell granuloma (CGCG), giant cell tumor (GCT), brown tumor (BT) and cherubism (Ch). Histologically all of the giant cell lesions appear similar, if not identical, and they usually can be distinguished by clinical history, immunohistochemistry or genetic markers. During ten years period at the Clinic for Maxillofacial Surgery of the Military Medical Academy there were 29 patients with lesions containing giant cells: 22 PGCG, one CGCG, three GCT, two BT and one cherubism. All patients with PGCG were successfully treated by single wide surgical excision of exophitic gum tumor and ostectomy of underlying bone. CGCG appeared in a male mandible. It has been removed together with adjacent bone, with no relapse. Two of three cases of the GCT of the mandible appeared in women of the fifth decade and after one local relapse they died of lung metastases within three years time; the third case is a 14 years old boy with GCT of maxilla: during the last three years he went through the parcial, and ten moths later the subtotal maxillectomy and he is in a stable remission for almost two years. Both young men with terminal renal failure and BT of maxilla were successfully operated. The 16-year old boy with cherubism was only diagnosed by bone biopsy. Conclusion: Although these lesions are histologically similar, their precise distinction is important because of their different biologic behavior and treatment. PP4-182 OUR EXPERIENCE WITH LOCALIZED CASTLEMAN'S DISEASE AT THE NECK Zoran Mirkovic, Slobodanka Vukelic-Markovic, Ruzica Kozomara, Zoran Damnjanovic, Nebojsa Jovic Clinic of Maxillofacial Surgery, Military Medical Academy, Serbia Background: Castleman's disease (angiofollicular lymph node hyperplasia or giant lymph node hyperplasia) appears as localized or ? rare - multicentric benign lymph node mass of the mediastinum, but it has been reported in abdominal anad retroperitoneal cavities, lungs, axillary and cervical region. Case report: during last 16 years there were four cases of Castleman's disease localized at the neck: 54 years old lady and three males: 20, 32 and 37 years old. All of them had asymptomatic, slowly growing tumor mass; diameter 9, 7, 8 and 9 cm respectively; without data about previous inflammatory episodes; under the upper part of the sternocleidomastoid muscle, with clinical presentation resembling lateral branchial cyst and clear ultrasonographic description of a solitary, clearly limited lymph node with abundant vascularization. Intraoperatively they were soft, fatty, bloody and easy removable. After pathohistologic verification of hyaline-vascular type of Castleman's disease in all three cases, patients are followed up during a period of 16, 5, 2 and one year with repeated ultrasonographic examination of the neck, axillas and abdomen and CT examination of the chest, but none of them developed any sign of local or multicentric relapse. Conclusion: Castleman's disease is the rarest benign lymph node hyperplasia in our surgical praxis. These four cases were successfully operated and carefully followed up for years. As we expected, none of them turned into multicentric form. PP4-183 ANALYSIS OF THE POSTOPERATIVE AND POSTIRRADIATION RELAPSE RATE OF INTRAORAL PLANOCELLULAR CARCINOMA Ruzica Kozomara, Slobodanka Vukelic-Markovic, Nebojsa Jovic, Srboljub Stosic Clinic of Maxillofacial Surgery, Military Medical Academy, Belgrade, Serbia Frequency and importance of etiologic factors in onset of oral squamocellular carcinoma (OPCC) at tongue and the floor of the mouth, together with their pathohistologic and clinical features,

point to high incidence of its relapse. According to available prognostic parameters, including tumor parameters (pathohistologic and TNM classification), it is not possible for certain to explain the disease relapse in patients with the same histologic type, stage and size of tumors which were treated in the same way (surgery and postoperative irradiation) and which were all pathohistologicly proved to be completely excised. During three years long clinical follow up of 84% male and 16% female patients, 43-80 years old, with OPCC of tongue and the floor of the mouth, we analyzed 12% patients in the second and 88% patients in the third stage of the disease, with 10% of T-1, 76% of T-2 and 14% of T-3 tumor size. The lesion was mostly localized at the middle third of tongue margin. 54% of patients showed the infiltrative tumor growth. All of the patients underwent tumor excision as well as regional lymphadenectomy. In 88% patients the dissection of the neck was performed: in 80% of patients it was suprahyoid ? 63% unilateral and 17% bilateral ? dissection, while 14% of the patients underwent radical and 7% supraomohyoid dissection of the neck. Pathohistologic examination confirmed positive lymh nodes in 71% cases of suprahyoid and all cases of radical and supraomohyoid dissection. Clinical follow up demonstrated relapse of the disease in 46% of the patients: in 65% of them it appeared early (during the first twelwe months after treatment), while in 35% of them in the period 12-36 months after treatment. Locoregional relapse appeared in 13% of the patients, but only as a late lesion. Simultaneous appearance of both local and locoregional relapse appeared early in 21% and lately in 8% of the patients. PP4-184 EXPERIENCE WITH LARGE DENTIGEROUS CYSTS OF THE MANDIBLE Slobodanka Vukelic-Markovic, Ruzica Kozomara, Zoran Mirkovic, Nebojsa Jovic, Srboljub Stosic Clinic of Maxillofacial Surgery, Military Medical Academy, Belgrade, Serbia Mandibular odontogenic cysts are common, but could be challenging when exceed more than 5 cm of mandibular lenght. During last ten years there was 100 large mandibular cysts: 40 developmental (25 dentigerous, 12 keratocysts, 1 eruption, 2 calcifying odontogenic cyst) and 60 inflammatory (59 radicular and 1 residual cyst). There was 67 males and 33 females; 14-62 years old (mean age 36 years). Most of cysts involve angular region (37), mandibular body (31), ascendent ramus (3), symphysis (7) and two or more of these regions (22). Two patients had two different cysts, and two patients simultaneously had some other intraoral lesion: fibroma and hemangioma. More than half caused external visible assimetry, pain and drainage. All of them were enucleated and residual defect has been reconstructed with aloplastic material (Osteovit), rarely marsupialized (8) or mandibula enhanced with metallic plate over remaining buccolabial cortex. In three cases reoperation was needed after two-seven years. In two cases of keratocyst nevoid basal cell carcinoma syndrome (Gorlin syndrome) has been present: 59 years old woman (keratocyst found after several excisions of facial basalioma) and 35 years old man (keratocyst operated 12 years before occurence of multiple facial basaliomas); both with epidermal cysts, enlarged head circumference, rib anomalies and mild hypertelorism, as well as pectus excavatum at male. PP4-185 PATHOHYSTOLOGIC ANALYSIS OF EFFECT OF CONCENTRATED PLATELETS ON OSTEOINTEGRATION OF DENTAL IMPLANTS Zoran Mirkovic, Zoran Lazic, Slobodanka Vukelic-Markovic, Smiljana Matic, Marija Bubalo Clinic of Maxillofacial Surgery, Military Medical Academy, Belgrade, Serbia

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INTRODUCTION: Biointegrity of an implant is based on expectation of their integration after placement within bony tissue by processes of remodelation and osteogenesis, so they could overtake functional load of the suprastructure. Aiming to create better conditions that influence to bone healing of dental implant, last few years growth factors are used in implantology. They are mediators of direct or indirect growth regulation of cells and tissues. Growth factors within CT-PRP important for bone regeneration are PDGF, TGF?, VEGF, EGF and other. AIM of this paper is to establish whether the application of CT-PRP method provides better osteointegration of dental implants. METHODS: The study was performed on six experimental dogs in which 24 BCT dental implants were placed: each got four implants ? two on the left side with CT-PRP and two on the right side without CT-PRP application. Osteotomized segments of the mandible were taken after 42, 70 and 98 days, and after preparation all were pathohistologically analyzed. Contact region of the implant with adjacent tissue has been analyzed pathohistologically by following of characteristic features of tissue reaction: connective tissue, osteoblasts and necrosis presence. RESULTS: Results obtained by pathohistological analysis demonstrated increase of connective tissue, outstanding osteoblastic activity and absence of necrosis in group where CT-PRP was applied, and increase of connective tissue, poor osteoblastic reaction and presence of necrosis during the first six weeks in group of implants placed in classic manner. CONCLUSION: According to results obtained from pathohistologic analysis the CT-PRP method has an advantage thanks to better osteointegration due to distinct osteoblastic reaction. PP4-186 PRIMARY EXTRANODAL NON-HODGKIN MALIGNANT LYMPHOMAS OF THE HEAD AND NECK REGION ACCORDING TO THE WHO CLASSIFICATION Yordanka Brachkova Department of Pathology, Medical University Pleven, Bulgaria BACKGROUND: The Working formulation commonly used to classify non-Hodgkin’s lymphoma (NHL) in Bulgaria, has been recognized as imperfect for primary extranodal lymphoma in head and neck regions. PURPOSE: To study the clinicopathological and immunohistochemical features of extranodal NHL according WHO Classification. METHODS: Routine histology was performed and stained with H&E and immunohistochemistry, and clinical characteristics were recorded. RESULTS: Between 1997 and 2006, 22 patients with primary extranodal NHL of head and neck region were treated surgically.The patients included 12 male and 10 female; age 24-80 years; tumor localization: tonsil ( n=10, 46%) sinunasal areas( n=4, 1.8%) nasopharynx ( n=2, 0.9%), salivary glands 2, floor of mouth/gingiva 2, hypopharynx 2. Immunohistochemically, 20 tumors were of B- cell phenotype and 2 tumors were of T-cell phenotype. According to the WHO classification , 12 cases were diffuse large B-cell lymphomas ( 8 in tonsil,2 sinonasal ), 3 were extramedulary plasmocytoma (paranasal sinuses-2, gigiva-1),1 was precursor B-lymphoblatic lymphoma (tonsila), and 1 was marginal zone B-cell lymphoma of mucosa associated lymphoid tissue ( MALT), 1 was mantle cell lymphoma,1 was Burkitt lymphoma. Among the T-cell lymphomas, 1 was peripheral T-cell lymphoma (tongue) and 1 extranodal NK/T-cell lymphoma (epipharynx). CONCLUSION: Our date correspond with series from Western population, but there is a significant difference from Japan, Hong Kong and Korea

PP4-187 HER-2 OVEREXPRESSION AND GENE AMPLIFICATION IN SALIVARY DUCT CARCINOMA Aleksandra Kolaric, Mats G Karlsson Dept of Pathology, Örebro University Hospital, Sweden Background Her-2 receptor overexpression in salivary duct carcinomas (SDC) was described in 1994. In the management of the look-alike tumour in the breast, Her-2 targeted therapy has been introduced and the evaluation of Her-2 receptor and gene status has been standardized and correlated to drug effects. In SDC, reports have confirmed Her-2 overexpression and gene amplification has been described. However, a variety of procedures has been used in these studies. Method 31 SDC were identified from the institutional archive, 28 of these were available for further analysis, including 8 of the 9 cases in the report of Hellquist. One block was stained and scored according to the Herceptest protocol (Dako, DK). Fluorescence in situ hybridization (FISH) including a Her-2 probe and a centromere 17 probe was performed and evaluated in 20 cells (Vysis, USA). A ratio (Her-2:centromere) of 2.0 was considered amplified. Cases with a high, not countable, signal were considered as clusters. Results Herceptest was negative (score 0) in twelve cases, 2+ in three and 3+ in eleven. All negative tumours had a normal genotype with a ratio 1.4. Of the three cases with score 2, one showed a normal FISH pattern, one low level amplification with ratio 2.2, whilst the third showed Her-2 cluster. All eleven cases with Herceptest 3+ showed gene amplification as clusters. In two cases a more than two-fold increase in centrome signals, one as clusters, indicated a more complex genotype. Two cases with intratumour heterogeneity were found, score 0/2+ and 0/3+. FISH was negative in Herceptest negative areas. 2+ area showed increased number of Her-2 as well as centromer signals indicating aberrations at chromosomal level, 3+ area showed clusters. Conclusion We conclude that Her-2 overexpression occurs more frequently in SDC than in ductal carcinoma of the breast. In breast carcinomas Her-2 gene amplification is considered the dominating mechanism for Her-2 overexpression and a prerequisite for targeted therapy. In SDC, a high frequency of overexpression in spite of a normal gene copy has been reported. In our series, no case showed overexpression at the 3+ level with a normal pattern for Her-2 gene signals. Gene amplification is the dominating mechanism for Her-2 overexpression also in SDC whilst a few cases with more complex genomic aberrations occur. The data imply that SDC is a potential target for anti-Her2 therapy and that Her-2 status should be evaluated with the same algorithm as in breast carcinomas. PP4-188 PRIMARY LEIOMYOSARCOMA OF THE THYROID GLAND Cigdem Celikel1, Murat Sari2, Burcu Tuzuner1, Mine Yazici2, Naziye Ozkan1 1 Marmara University School of Medicine, Department of Pathology, Turkey 2 Marmara University School of Medicine, Department of ENT, Turkey Primary leiomyosarcoma of the thyroid gland is extremely rare and clinico-pathological information on this tumor is scant. We herein report a case of epithelioid leiomyosarcoma of the thyroid gland in which anaplastic carcinoma was suspected based on clinical and cytological features. The patient was a 79-year-old male who presented with a painful mass in the left neck region that had increased in size over a number of months. Ultrasonography revealed a mass lesion in the left lobe of the thyroid gland. Fine needle aspiration was performed. The cytological diagnosis was a malignant tumor compatible with carcinoma. In the surgical specimen of the thyroid left lobe and isthmus, there was a mass lesion measuring 6 cm in its greatest

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dimension that was fleshy, necrotic and uncapsulated. Histologically, there was a solid, lobular pattern of growth comprised of epithelioid cells with increased cellularity, pleomorphism, a high mitotic rate, necrosis, and hemorrhage. Immunohistochemistry of the tumor cells clearly showed smooth muscle differentiation; the cells were positive for desmin, muscle-specific actin and vimentin and negative for cytokeratin, epithelial membrane antigen, carcinoembryonic antigen, thyroglobulin and calcitonin. The tumor cells also showed positive reactivity to c-kit proto-oncogene product. There was no evidence of metastasis in the radiological and clinical examination. No local recurrence and metastasis developed within 4 months after the initial operation. Leiomyosarcomas of the thyroid gland are distinctive tumors and can be distinguished from anaplastic carcinoma with the aid of immunohistochemistry. PP4-189 HPV-RELATED SQUAMOUS PAPILLOMA OF THE OROLARYNGEAL TRACT. REPORT OF TWO CASES FROM THE TONSILS AND THE LARYNX Maria Papaevangelou1, Efthymios Koniaris1, Nikolaos Zirganos1, Vasileios Negris1, Nikiforos Kapranos2, Nikiforos Apostolikas1 1 Pathology Dpt., Anticancer Oncological Hospital of Athens ‘St. Savvas’, Athens, Greece 2 Molecular Pathology Dpt., ‘Mitera’ Maternity and Surgical Center, Athens, Greece Background: A variety of HPV-related lesions have been reported in the oral cavity and in the larynx. A wide spectrum of architectural patterns ranging from simple Epithelial Hyperplasia (EH), to Verruca Vulgaris (VV) and from Condylloma Acuminatum (CA) to Squamous Papilloma (SP), characterizes these lesions. Here we report two cases of SP, one from the larynx and one from the tonsil. Method: The first case was traced in a 45-year old sailor who referred to our hospital for a sore throat and endoscopic removal of a small laryngeal, warty lesion took place. We received 6 tissue specimens measuring 0.3-0.5cm. The clinical history of the patients wife, revealed an HPV infection of the cervix. The second case involved a 26-year old female with a small papilloma measuring 0.5cm on her right tonsil. Results: Histologically, both lesions were warty-papillary with branching projections consisted of squamous epithelium overlying thin fibrovascular cores. The basal, parabasal cell hyperplasia, the mild anisonucleosis and the non-uniform perinuclear halos of both lesions, motivated us to screen the positivity with the different subtypes of the papilloma viruses. Immunohistochemistry revealed strong staining for HPV types 6 and 11 in both cases. Conclusion: The SPs are the most common benign laryngeal tumors (84%) caused by HPV. The adult-onset oral and laryngeal SP is a frequently relapsing disease. HPV types 6 and 11 which do not carry a high risk of malignant transformation have been detected with a variable frequency (8-60%). Virus transmission appears to be mostly horizontal or by autoinoculation. These patients have a history of a genital wart in 16% and their female sex partners 12.5%. Primary HPV infection may remain latent, requiring cofactors to develop into the clinical disease. A lack of orogenital contact does not protect the patients from adult-onset SP since they seem to be prone to HPV infections. Differential diagnosis involves kerstinized SP, CA and verrucous carcinoma. The diagnosis plays an important role for the appropriate treatment since the relapse is very common. Therapy in laryngeal papillomatosis involves 6 sessions of laser ablation of the papillomas and intralesional injections with cidofovir.

PP4-190 ATYPICAL LYMPHOCYTIC INFILTRATION (LYMPHOCYTOMA) OF THE INFERIOR ALVEOLAR NERVE: A CASE REPORT Maria Siponen1, Veli Matti Vartiainen2, Tuula Salo1, Jukka Rosberg1, Meeri Apaja-Sarkkinen3 1 Institute of Dentistry, University of Oulu, Finland 2 Department of Diagnostics and Oral Medicine, Oulu University Hospital, Finland 3 Department of Pathology, Oulu University Hospital, Finland Lymphocytomas are non-neoplastic processes occurring mainly in the skin that are characterised histopathologically by polyclonal lymphocytic infiltrate. They may be associated with Lyme’s borreliosis but in many cases etiology remains unclear. Differential diagnosis includes also small cell malignant lymphomas. 33-year-old male was referred to the Department of Oral and Maxillofacial Surgery at Oulu University Hospital for gradually developed sensory loss of the left lower lip. Radiological examinations (panoramic x-ray, CT-scan and MRI) revealed enlarged left mandibular canal, with an expansive, but well-defined soft tissue lesion. In the bone scan a local accumulation was seen in the left body of the mandible. A biopsy was taken from the inferior alveolar nerve. Histopathological findings suggested a lymphocytic infiltration of the nerve tissue. Immunohistochemistry showed both B- and T-lymphocytes in the infiltrate. Some of the B-lymphocytes were Bcl-2 and CD23 positive, which was considered to be related to residual mantle zones. Later the whole enlarged section of the inferior alveolar nerve was resected, and showed similar histopathological findings. Further medical examinations have not revealed any systemic disease and the patient has been well in the 16-month follow-up period. PP4-191 THE IMPORTANCE OF HYALURONIC ACID IN VOCAL FOLD LESIONS IN DIFFERENTIAL DIAGNOSIS Nil Comunoglu1, A. Isin Dogan Ekici1, Ferda Ozkan1, Ismail Kocak2, Bulent Eren3, Selcuk Bilgi1 1 Yeditepe University, School of Medicine, Department of Pathology, Turkey 2 Yeditepe University, School of Medicine, Department of ENT, Turkey 3 Uludag University, School of Medicine, Department of Forensic Medicine, Turkey Background: We examined the influence of hyaluronic acid and basement membrane thickness alterations on biochemical properties of vocal fold lesions including vocal fold polyps, sulcus vocalis lesions, vocal fold cysts, and Reinke’s edema. Method: Fifty-six laryngoscopic vocal fold biopsies from 49 patients, reported between 1.7.2005 and 27.2.2007 in Yeditepe University, School of Medicine, Department of Pathology, were evaluated retrospectively. Twenty-seven of these cases were vocal fold polyps, 7 were sulcus vocalis, 13 were vocal fold cysts, and 9 were Reinke’s edema. Ten normal vocal fold mucosa samples from autopsy cases were included in the study as control group. In order to evaluate the hyaluronic acid in extracellular matrix and basement membrane alterations; hematoxylin-eosin, alcian blue pH 2.5, PAS, and colloidal iron stains were performed histochemically. We evaluated the staining pattern for each lesion and compared with that of control group. Data analysis was performed using SPSS Version 8.0 (Chicago, IL) statistical package. Frequency data were evaluated by the Kruskall Wallis test. A P value 0.01 was considered to be statistically significant. Results: We detected that sulphated hyaluronic acid in extracellular matrix was increased with neomatrix formation in vocal fold polyps, sulcus vocalis lesions, vocal fold cysts, and Reinke’s edema when compared with control group. The difference was statistically significant (p<0.01). This increase was more remarkable in vocal fold polyps than sulcus vocalis

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lesions, vocal fold cysts, and Reinke’s edema. It was detected that basement membrane thickness increased in 4 pathologic lesion groups compared with control group in statistically significant manner (p<0.01). This increase in basement membrane thickness was most remarkable in sulcus vocalis lesions. Conclusion: The results suggested that hyaluronic acid plays an important role in determining biochemical properties of vocal fold lesions including vocal fold polyps, sulcus vocalis lesions, vocal fold cysts, and Reinke’s edema. Furthermore we think that hyaluronic acid and basement membrane thickness alterations can be useful in differential diagnosis in vocal fold lesions. PP4-192 THE RELATIONSHIP BETWEEN p63 EXPRESSIONS IN BASALOID AND CONVENTIONAL SQUAMOUS CARCINOMAS OF THE LARYNX Semsi Altaner, Omer Yalcin, Fulya Oz Puyan, Ufuk Usta Trakya University, Medical Faculty, Department of Pathology, Edirne, Turkey INTRODUCTION: p63 is a p53 homologue that seems to play distinctive roles in the physiology of the squamous epithelia. p63 is similar to p53 but it has different activities. Our aim was to obtain new insights into the role of p63 in basaloid squamous cell carcinoma (BSCC) and conventional squamous cell carcinoma (CSCC). MATERIALS and METHODS: We examined the presence of p63 in 20 BSCC and in 20 CSCC cases retrospectively at immunohistochemical level. The expression of p63 was analyzed in paraffin-embedded specimens of 20 patients with BSCC and 20 patients with CSCC by using immunohistochemical staining methods. RESULTS: All cases with BSCC and CSCC stained positively with p63. Diffuse staining rates with p63 in the BSCC was significantly different from the ones in the CSCC group (p<0.05). 11 cases in the basaloid group and 4 cases in the conventional group showed diffuse staining with p63. Diffuse staining with p63 was evident on the sections from the specimens with poor prognostic factors in the basaloid group. CONCLUSION: In the present study, we determined p63 positivity in the all BSCC and CSCC cases of the larynx. In addition, diffuse staining pattern with p63 in the BSCC cases may have a value in distinguishing between the CSCC and BSCC. We suggest that p63 diffuse positivity has a relationship with the prognostic parameters and it seems to be a poor prognostic factor in BSCC of the larynx. PP4-193 HR-CGH IN BASAL CELL ADENOMAS AND ADENOCARCINOMAS Fabricio Passador-Santos1, Sara Martorelli2, Suzana Sousa3, Vera Araujo4, Fernando Soares5, Luiz Kowalski6, Silvia Rogatto2 1 University of São Paulo -USP - School of Dentistry - Department of Oral Pathology, Brazil 2 São Paulo State University - UNESP - School of Medicine - Neogene Laboratory, Brazil 3 University of São Paulo - USP - School of Dentistry - Department of Oral Pathology, Brazil 4 São Leopoldo Mandic - School of Dentistry, Brazil 5 AC Camargo Hospital - Department of Pathology, Brazil 6 AC Camargo Hospital - Department of Head and Neck Surgery, Brazil Background: Salivary gland tumors (SGT) are rare neoplasms comprising from 2 to 4% of all head and neck tumors. The overall low incidence and histological diversity are major limitations for all biological studies on the salivary gland tumors. The molecular events associated with their development and clinicopathological heterogeneity remain unknown. Previous cytogenetic and molecular genetic analysis of these tumors have been limited in scope and size and did not account for their inherent morphological and biological heterogeneity. Basal cell adenomas (BCAs) and basal cell adenocarcinomas (BCACs)

represent, respectively, the benign and the malignant counterpart of salivary gland tumors characterized by the proliferation of basaloid cells. Histologically, they can be very similar, especially when a few amount of material is available for analysis, such as incisional and fine needle biopsies. Method: High resolution comparative genomic hybridization (HR-CGH) using metaphase chromosomes can detect changes in chromosome copy number with a resolution of 3-20Mb. We applied HR-CGH technology in 10 cases (from nine patients) of basal cell adenomas and 5 cases (from four patients) of basal cell adenocarcinomas in order to evaluate and to compare the gains and losses profile in both tumors. Results: Interestingly, it was detected a characteristic pattern of gains and losses in both group of tumors. Losses in 4p14-p15.1, 10q25-q26, 14q24-q31 and 19q13.2-q13 and gains in 22q13 were found in the BCACs. On the other hand, BCAs showed gains in 2q37, 3p24-p26, 4q33-q35, 5q23-q31, 7p22, 10q25-q26, 11p15, 11q25, 15q26, 18p11.3 and 20q13.2-q13.3. Two samples from the same patient (BCAC2 and BCAC5) showed different copy number alterations. In this case, one sample shared the same pattern of chromosomal alterations observed in the basal cell adenomas. Conclusion: In overall, the results support the hypotheses that a set of genes differentiate BCAs and BCACs. These results provide new information on potential genetic events of biological significance in future studies of these salivary gland tumors. PP4-194 IDENTIFICATION OF METASTATIC ADENOCARCINOMA AND REACTIVE MESOTHELIAL CELLS IN PLEURAL EFFUSIONS. VALUE OF AN IMMUNOCYTOCHEMICAL PANEL OF CD44, CALRETININ, CK7, CK20 AND TTF-1 Nur Yucel1, Zekiye Aydogdu Dinc1, Ali Kadri Cirak2 1 Izmir SS Chest Disease and Surgery Hospital, Department of Pathology, Turkey 2 Izmir SS Chest Disease and Surgery Hospital, Clinic of Chest Disease, Turkey Background: Pleural effusions are frequently first clinical manifestation of metastatic disease. Lung, breast, ovarian carcinomas and malignant mesothelioma (MM) are leading the causes of malign pleural effusions. Cytologic differentiation of metastatic carcinoma and reactive mesothelial cells in effusion is sometimes a problem that cytomorphologic criters are insufficient. Recently, extensive research has resulted in expansion of the antibody panel that is available for effusion diagnosis, thereby reducing the risk for error Methot: We have used a panel of five monoclonal antibodies; Calretinin, CD44, CK 7, CK 20 and TTF-1, so as to identify the primary tumor site of metastatic carcinoma cells and reactive mesothelial cells in pleural effusions. Results: Applying an algorithm of immunocytochemical marker constellations, we were able to correctly diagnose proliferating mesothelial cells (% 100) and primary tumor sites in 23 of 30 patients. The best result was achieved for the identification of metastatic carcinomas of the lungs (88.1%).We established an algorithm comprising five immunocytochemical markers that enabled a correct diagnosis of primary tumor sites in 75%. Conclusion: The panel studied could be useful in diagnostic routine for the identification of primary tumors of unknown origin, metastatic to the pleural membranes and reactive mesothelial cells.

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PP4-195 p16 EXPRESSION IN HUMAN PAPILLOMAVIRUS RELATED PREMALIGNANT HEAD AND NECK LESIONS S. Elif Gultekin1, J. Peter Klussmann2, Soenke Weissenborn3, Hans. P. Dienes4 1 Gazi University Dental Faculty Department of Oral Pathology, Turkey 2 University of Cologne Department of Oto-Rhyno-Laryngeology, Germany 3 University of Cologne Virology Institute, Germany 4 University of Cologne Pathology Institute, Germany

Background: Human papillomavirus (HPV) induced orophraynx carcinomas has been postulated as a distinct tumor entity in terms of biological behavior and treatment modality. p16 is a tumor supressor gene product ,which is a strong candidate of a valuable prognostic marker ,shown to be overexpressed in most oropharangeal carcinomas associated with high risk HPV. Material and Methods: The study was conducted on paraffin embedded specimens of 43 cases of tonsillar and laryngeal lesions which comprised 15 cases of dysplasia and 28 cases of papilloma. Eleven of 15 dysplasia cases were from tonsils and 4 of 15 cases were from larynx. Most of the tonsillar dysplasia specimens were from the dysplastic epithelium adjacent to tonsillar carcinoma (10 of 11 cases). Twenty four papilloma cases were from tonsils (14) and larynx (14). p16 expression was evaluated by ABC immunoperoxidase staining whereas the presence subtypes of HPV DNA was determined by polimerase chain reaction (PCR) . All dysplasia cases were graded and p 16 positivity was graded as high, medium, low degree (score1-2) on the basis of staining pattern and density. Results: Seventeen of 43 cases (39.5 %) showed the presence of HPV DNA. Of positive specimens, 7 of 17 (41.2 %) were papilloma and 10 of 17 (58.8 %) were dysplasia lesions. All HPV positive papilloma specimens were from larynx . Of positive dysplasia cases, 9 of 10 were tonsillar and 1of 10 was larynx lesions. Nine of 10 dysplasia cases adjacent to carcinoma showed HPV 16 positivity, where as 1 laryngeal dysplasia case had low risk viral load (HPV 6). All HPV positive laryngeal papilloma cases showed the presence of low risk HPV DNA load (HPV 6/11). High degree (score 3) p 16 overexpression was seen 12 of 15 (80 %) dysplasia and 1of 28 (3.6 %) papilloma cases. All HPV 16 positive dysplasia cases had high degree p 16 over expression,. Conclusion: The results showed HPV was not detected in tonsillar papillomas while HPV 11 /6 was found half of the larynx lesions. High risk HPV (HPV 16) was detected in tonsillar dysplasia lesions and showed positive correlation with p16 overexpression. PP4-196 MIDDLE EAR CHOLESTEATOMA: CORRELATION BETWEEN IMMUNOHISTOCHEMICAL FINDINGS AND DISEASE EXTENSION AND BONE DESTRUCTION Jovan Dimitrijevic1, Nada Milanovic2, Miodrag Colic3, Vera Todorovic4, Neda Drndarevic4 1 Institute of Pathology, Military Medical Academy, Belgrade, Serbia 2 Institute of Otorhinolaryngology, Military Medical Academy, Belgrade, Serbia 3 Institute for Medical Research, Military Medical Academy, Belgrade, Serbia 4 University of Belgrade - Institute for Medical Research, Department of Immunohistochemistry and Electron Microscopy, Belgrade, Serbia

Background: Cholesteatomas are accumulation of exfoliated keratin inside the middle ear, originated from keratinized squamous epithelium. These epidermoid cyst show independent and progressive growth with destruction of adjacent tissue, especially the bone tissue, with tendency to recurrence. According to some investigations, a characteristic signal of cholesteatoma is the infiltration in the perimatrix of immune

system cells, and the increment in proliferation of the cholesteatoma matrix would be the results of the inflammatory process, suggesting that the perimatrix would be the main factor of cholesteatoma development. Method: The purpose of the current study was to analyze the immunohistochemical features of cholesteatoma in correlation with extension and bone destruction grades. Specimens were obtained from 30 patients (9 female and 21 male; mean age, 46 years) requiring middle ear surgery. To define the cell density and distribution of T and B cells, CD4+ and CD8+ cells, antigen-presenting cells, macrophages, Mercels cells, Langerhans cells, mastocytes, as well as expression of E-cadherin, we used CD3, CD4, CD8, CD68, CD1a, CD117, HLA-DR, triptase, CD19, CD38 and E-cadherin antibodies and ABC immunohistochemical procedures. Results: Generally, immunostaining suggests that the T cells (CD4+>CD8+) and antigen-presenting cells (HLA-DR+) are predominant cell populations in the cholesteatoma, as well as that triptase+ mastocytes more number in comparison to CD117+mastocytes. Langerhans cells (CD1a+) are often seen intraepithelialy. There was increase density of each CD3+, CD4+, CD68+ cells in cases of cholesteatoma with extensive erosion and sequestration of the underlying bone. In addition, it is evidence of increased number of intraepithelial CD8+ cells in same cases. No statistical differences in density of each HLA-DR+, CD1a+, CD117+/triptase+, CD19+, CD38+ cells was found between cholesteatoma with and without extensive erosion of the underlying bone. Also, results show that the expression and distribution of E-cadherin in middle ear cholesteatoma is not deranged with extensive bone erosion. Conclusion: It is well known that the progression of the cholesteatoma might be induced by the release of factors from the cholesteatoma matrix or perimatrix. Findings of our study suggested that cholesteatoma CD3+, CD4+, CD8+ and CD68+cells plays and active role in the process of bone destruction. PP4-197 THE EFFECT OF CAPECITABIN OR 5-FLUOROURACIL (5-FU) ON PREVENTION OF EPISCLERAL FIBROSIS AFTER TRABECULECTOMY IN THE RABBIT – TWO WEEKS FOLLOW-UP PATOHISTOLOGICAL FINDINGS Milorad Milivojevic1, Jovan Dimitrijevic2, Miroslav Vukosavljevic1, Petar Aleksic1, Katarina Jelic2, Zoran Latkovic3 1 Clinic of Ophthalmology, Military Medical Academy, Belgrade, Serbia 2 Institute of Pathology, Military Medical Academy, Belgrade, Serbia 3 Institute of Ophthalmology, Clinical Center of Serbia, Belgrade, Serbia

Background: The fundamental role of the immune system in conjunctival wound healing after glaucoma surgery is well established. Glaucoma filtering surgery fails most frequently due to fibrosis at the episcleral-conjunctival/Tenon’s capsule interface. The success rate of glaucoma filtration surgery may be optimized through the use of a combination of pharmacological agents that modulate the inflammatory and proliferative phases of the wound healing reaction. The most commonly clinically used drugs are corticosteroids and the antimetabolites, including 5-FU. Currently, many new pharmacological agents are used to modulate the wound healing. Method: In a rabbit model of glaucoma, we were examined the effects of capecitabin, a 5-FU Prodrug, or 5-FU on prevention of episcleral fibrosis after trabeculectomy. Twenty four animals were included in the study. After trabeculectomy we placed 5% solution of 5-FU in one eye of each animals for two weeks and the fellow eye was received capecitabin. Eye specimens were investigated by histological examination. Results: Microscopic examination revealed no statistical significant change in the number of fibroblasts and granulocytes, as well as collagen fibers density in the episclera between experimental groups. However, we observed the significant decrease of number of mononuclear cells and

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plasmocytes, and reticulin fibers density in the eyes treated with capecitabin. Conclusion: Although both 5-FU and capecitabin were effective in the prevention of episcleral fibrosis after trabeculectomy in the rabbit, capecitabin has stronger effect in suppression of chronic inflammation. PP4-198 EFFECT OF TREPONEMA DENTICOLA ON HUMAN DENDRITIC CELLS PHENOTYPE AND FUNCTION Patrıcıa Cury1, Veronica Horewicz1, Joao Carmo2, Vera Araujo1, Ney Araujo2, Jose Barbuto2 1 Sao Leopoldo Mandic Dental Research Institute, Brazil 2 University of Sao Paulo, Brazil

BACKGROUND: Periodontitis is a bacterial infection characterized by chronic gingival inflammation, which leads to the loss of the tooth-supporting tissue. Dendritic cells play an important role as regulatory and effector cells in the pathogenesis of periodontitis and Treponema denticola is an important periodontopathogen. This study compares the effect of Treponema denticola on the phenotype and function of human dendritic cells obtained from peripheral blood mononuclear cells of volunteers with periodontal healthy or chronic periodontitis. METHODS: Peripheral blood was obtained from systemically healthy volunteers with periodontal healthy or chronic periodontitis. Monnonuclear cells were isolated and maturated in dendritic cells. Dendritic cells were pulsed with extract of Treponema denticola at 5 g/ml, 10 g/ml or 15 g/ml, irradiated and were then cocultured with allogeneic T cells for 7 days. Dendritic cells were marked with monoclonais antibodies anti-CD1a, anti-CD14, anti-CD80, anti-CD86, anti-CD83, anti-CD11c, anti-CCR7, anti-HLADR. RESULTS: In periodontal healthy volunteers the bacteria increased the maturation of the dendritic cells, while in the periodontitis volunteers, the maturation was decreased when HLADR and CD-11c were analyzed. Proliferation of T cells was higher in the healthy than in the periodontitis volunteers. CONCLUSION: Phenotype and function of dendritic cells from individuals with periodontal healthy and chronic periodontitis differs when dendritic cells are pulsed with Treponema denticola. This may explain at least in part the differeces in susceptibility to periondontitis. PP4-199 CYTOKERATIN EXPRESSION PATTERNS OF PRECANCEROUS LESIONS OF THE LARYNX CLASSIFIED ACCORDING TO THE LJUBLJANA AND WHO CLASSIFICATIONS Yesim Gurbuz1, Evrim Kus1, Omer Aydin2 1 Kocaeli University Medical Faculty Pathology Department, Turkey 2 Kocaeli University Medical Faculty Otorhinolaryingology Department, Turkey

INTRODUCTION: Ljubljana system for the precancerous lesions of the larynx is claimed to be more predictive for malignancy than WHO system. The main difference between these two classifications is the high percentage of involved epithelium in high grade displasia in WHO classification that corresponds to atypical hyperplasia in Ljubljana system. In normal laryngeal mucosa cytokeratin 14 (CK14) expression in basal, ciliated and non ciliated epithelium, CK8, CK18 and CK19 existence in non basal cells and CK13 expression in squamous and cuboidal cells were reported. In this study, we examined the cytokeratin expression patterns of precancerous lesions of the larynx classified according to Ljubljana and WHO classification in order to compare these two systems and altered cytokeratin expression in laryngeal carcinogenesis. MATERIAL METOD: We included 1 selected block from 42 laryngeal biopsies in our study (2 simple hyperplasia, 18 abnormal hyperplasia, 15 atypical hyperplasia, and 7 carcinoma insitu according to Ljubljana classification), (1 normal mucosa, 12 mild dysplasia, 9 moderate diysplasia, 12 severe dysplasia and 8 carcinoma insitu according to WHO

classification). CK 7, 10, 13, 14, 16, 17, 19, 20 were detected by immunohistochemical metod. The relation between cytokeratin expression and subgroups were classified according to Ljubljana and WHO classification were detected by Chi square test. RESULTS In Ljubljana classification, CK13 expression was observed in 95% of simple hyperplasia, abnormal hyperplasia and 72.7% of, atypical hyperplasia and carcinoma insitu. CK16 expression is decreased with the intensity of the lesion (p<0.053). In WHO classification, CK10 expression was observed in 76.9% of normal mucosa and mild dysplasia, 48.3% of moderate, high dysplasia and carcinoma insitu (p<0.083). CK16 expression was detected in 76.9% of normal mucosa and mild dysplasia 41.1% of moderate, high dysplasia and carcinoma insitu (p<0.033). There was CK19 expression in 38.5% of the normal mucosa and mild dysplasia 75.9 % of moderate,high dysplasia and carcinoma insitu (p<0.019). CONCLUSION: WHO classification is more related with the cytokeratin expression than Ljubljana system. The reason may be the different levels of involvement of the squamous epithelium in two classification systems. There is increase in CK19 and decrease in CK10, 13 and 16 expressions in laryngeal carcinogenesis process. PP4-200 MUC1, MUC2, MUC4, MUC5AC, MUC6 AND PS2 EXPRESSION IN SALIVARY GLAND NEOPLASIAS Yesim Gurbuz1, Deniz Filinte1, Omer Aydin2 1 Kocaeli University Medical Faculty Pathology Department, Turkey 2 Kocaeli University Medical Faculty Otorhinolarngology Department, Turkey

INTRODUCTION: Mucins are high density molecules synthesized by epithelial cells. Thirteen different type of mucin is defined. MUC 1, MUC 2, and MUC 4 were observed in the salivary gland ductal system. MUC 6 expression has been found in normal and neoplastic salivary gland tissue. TFF-1 is the molecule which makes co-expression with MUC 5 AC and was detected in the salivary glands. MUC expression in salivary gland neoplasias is variable and not studied adequate. Salivary gland tumors are a group of the neoplasias with problematic differential diagnosis especially in small biopsies. In this study, MUC and TFF1 expression of a group of salivary gland neoplasia were examined in order to find an additional criterion for their differential diagnosis. MATERIAL METOD For this purpose, 41 salivary gland tumors (10 pleomorphic adenoma, 10 Warthin tumor, 10 adenoid cystic carcinoma, 5 mucoepidermoid carcinoma, 4 myoepithelioma and 2 basal cell adenoma ) have been included in the study. MUC 1, MUC 2, MUC 4, MUC 5 AC, MUC 6, and TFF-1 expressions were detected immunohistochemically. The presence and the intensity of the expression were scored as 0 to 4. Relation between existence of MUC 1, MUC 2, MUC 4, MUC 5 AC, MUC 6, TFF-1 expressions and MUC 5 AC and TFF-1 expression patterns ( MUC 5 AC +/TFF-1 +, MUC 5 AC -/TFF-1 -, MUC 5 AC +/TFF-1 -, MUC 5 AC -/TFF-1 + ) of different tumors, malignant and benign groups were detected by Chi-square test. RESULT: MUC 1 and MUC 4 expression was detected in all mucoepidermoid carcinomas, MUC 2 expression was detected in 4 cases (80%). MUC 6 existence in the mucoepidermoid carcinoma and adenoid cystic carcinoma was 20% and 40%. MUC 5 AC+/TFF-1 + staining pattern was observed in 62.9% of the benign neoplasias, and in 6.7% of the malignant salivary gland tumors (p< 0.0001). MUC 5 AC -/ TFF-1 - staining pattern was observed in 11.5% of benign and 53.5% of malignant salivary gland tumors (p < 0.004). CONCLUSION MUC 5 AC+/TFF-1 + staining pattern is more common in benign salivary gland neoplasias compared with the malignant group. Co expression of these two molecules in benign salivary gland neoplasias is similar to their normal expression in normal tissue. MUC-5 AC and TFF-1 expression may be an additional differential diagnostic parameter in salivary gland neoplasias.

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PP4-201 CHRONIC HYPERPLASTIC CANDIDIASIS OF THE LARYNX Ugur Pabuccuoglu1, Sulen Sarioglu1, Enis Alpin Guneri2, Cenk Ecevit2 1 Dokuz Eylul University Faculty of Medicine Department of Pathology, Turkey 2 Dokuz Eylul University Faculty of Medicine Department of Otorhinolaryngology, Turkey Background: Candida species induce chronic inflammatory changes in the laryngeal epithelium comparable to chronic hyperplastic candidiasis (candidal leukoplakia) of the oral mucosa. However, hyperplastic candidiasis of the larynx is assumed to be rare. We here report a prospective series of 15 patients with chronic laryngeal candidiasis and discuss the diagnosis and management of these lesions. Methods : The histopathologic diagnosis of candidiasis was based on morphological features of causative fungi as well as associated epithelial changes. Patients were treated by type I, type II or type III resections. Excisions were followed by antireflux medications and systemic flucanozole treatment and patients were advised to stop smoking . Results : Of 15 patients with chronic laryngeal candidiasis, twelve were men (%75) and 3 (%25) were women. The ages of the patients ranged from 36 to 82 years. Hoarseness was the most common presenting symptom (13 cases). In 11 cases the history of smoking and in three cases associated alcohol consumption was recorded. Clinical impression was that of a hyperkeratotic lesion in 10 cases. Histopathological features associated with candidal organisms in biopsy specimens included; epithelial hyperplasia, keratosis and parakeratosis of varying severity, and polymorphonuclear leukocyte infiltration at the superficial layers of the laryngeal epithelium. Candidal hyphae were often scarce and located within the keratotic layer. Epithelial invasion by the hyphae was limited to uppermost epithelial layers. Periodic acid Schiff stain was very useful for the detection of fungal organisms. Mild epithelial dysplasia was noted in 1 case and moderate dysplasia was encountered in 2 additional cases. Histopathologically, in 8 cases fungal hyphae involved right vocal cords while in 5 cases they were noted in the left vocal cords. In one case bilateral vocal cords were affected. In the remaining case candidiasis was associated with squamous cell carcinoma. Conclusions: The possibility of hyperplastic candidiasis should be kept in mind when managing cases of keratosis and leukoplakia of the larynx. The pathologist must be aware of histopathological changes associated with hyperplastic laryngeal candidiasis and should apply special stains in order to detect fungi. This may influence the low incidence of laryngeal candidal leukoplakia, which in fact seems to be relatively higher in our experience. Laryngopharyngeal reflux may be an important predisposing factor in the development of laryngeal candidal leukoplakia. PP4-202 DERMATOFIBROSARCOMA: CASE REPORT Zoran Mirkovic, Slobodanka Vukelic-Markovic, Nebojsa Jovic, Sasa Jovic Clinic of Maxillofacial Surgery, Military Medical Academy, Belgrade, Serbia Dermatofibrosarcoma is rare tumor in head and neck region, it is asymptomatic and it rarely metastasizes. Here we present a case of on extremely aggressive, rapidly growing and mutilating DFS seemingly non-responsive to wide surgical excision. 42 years old men underwent surgical excision of discreet, painless left cheek soft tissue lesion. There were no palpable lymph nodes on the neck and chest radiography was normal. On histopathological examination, the lesion was thought to be dermatofibroma. Three years later, he presented with a recurrence at the site of excision. After excision the recurrent lesion was suggestive of dermatofibrosarcoma. During almost next three years the patient

has been operated six times: each time the tumor rapidly and aggressively had grown outwards, destroying the whole middle third of the face, so at the moment of death only forehead, left eye, tongue and part of mandible had left. Comparing to dermatofibrosarcoma protuberans which appeared in our three patients, slowly relapsed and never tended to be aggressive, dermatofibrosarcoma itself, on the other hand, acted completely reverse, especially in this case where we have observed visible rapid growth of huge tumor masses destroying face, never giving regional or distant metastases. PP4-203 EXTRAMEDULLARY PLASMACYTOMA OF NECK T. Jakovina1, K. Jakovina1, D. Danic2, I. Pirkl2, K. Tomic1, Lj. Fustar-Preradovic1 1 Department for Pathology, Forensic Medicine and Cytology, General Hospital Slavonski Brod, Croatia 2 Department of Othorhinolaryngology and Head and Neck Surgery, Croatia Plasma cell neoplasms and related entities are a group of lymphoid neoplasms of terminally diferentiated B cells that have in common the expression of a single clone of Ig secreting plasma cells. Multiple myeloma is the most common disorder. It is characterized by multiple tumorous mases of neoplastic plasma cells scattered throughout the sceletal system. About 3-5% of plasma cells neoplasms present as a solitary lesion of either bone or soft tissue. Aproxymately 80 % of all extramedullary plasmacytomas occur in the upper aerodigestive tract, especially the sinonasal tract, and 20 % in the other body sites. CASE: We present an anusual case of extramedullary plasmacytoma located in the neck of a 73 years old man. The patient initialy presented with tumorous mass in the right side of the neck. Ultrasonography showed tumorous mass. Ultrasound –guided fine needle aspiration of that lesion had been performed. Cytologic differential diagnosis was plasmacytoma or oncocytic tumor. In bone marrow aspirates we have found 8% plasma cells. Tumor was removed and hystological analysis revealed tumor composed of sheets mature plasma cells with round eccentric nuclei and abundant cytoplasm. Imunohystochemical analysis shoved positive reaction for CD 138 and IgA and negative for CD 20, CD 3 and IgM. Hystological analysis and immunohistochemistry confirmed diagnosis of plasmacytoma. CONCLUSION: Extramedullary plasmacytomas comprises less than 1% of all head and neck tumors. Extramedullary plasmacytoma of the neck region is extremely rare. Treatment consists of surgically or/and radiation therapy with tipically good prognosis.

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Neuropathology PP4-204 EXPRESSION OF THE PARATHYROID HORMONE-RELATED PROTEIN IN GLIAL TUMORS Nagihan Yalcin1, Nilay Sen Turk1, Hulya Tosun1, Nese Calli Demirkan1, Bahar Baltalarli2, Erdal Coskun3 1 Department of Pathology, Pamukkale University School of Medicine, Denizli, Turkey 2 Department of Radiation Oncology, Pamukkale University School of Medicine, Denizli, Turkey 3 Department of Neurosurgery, Pamukkale University School of Medicine, Denizli, Turkey BACKGROUND: It has been found that Parathyroid hormone-related protein (PTHrP) is expressed in a variety of tumors including breast, prostate, colon, lung, renal and ovarian cancers. In some recent publications, it has been noted that there is a relation between PTHrP expression and prognosis in glial tumors. The aim of this study is to evaluate the expression of PTHrP in glial tumors and to find out whether there is a relationship between this expression and grade. METHODS: We have examined the expression of PTHrP in glial tumors of 16 glioblastoma multiforme , 2 anaplastic astrocytoma, diffuse astrocytoma grade II and pleomorphic xanthoastrocytoma grade II using immunohistochemical staining. RESULTS: It has been found that PTHrP staining has been seen in 6 glioblastoma multiforme, a pleomorphic xanthoastrocytoma. The strongest and the most diffuse staining has been seen in the case of pleomorphic xanthoastrocytoma. The patient was a child. It has also been found that staining is very weak in 4 out of the 6 fixed glioblastome. Four cases showed weak staining out of six positive gliobastome. CONCLUS ON: PTHrP is expressed very low in human glial tumors in our study. Further studies of large series having more low grade astrocytomas should be done. PP4-205 RARE OCCURRENCE OF OLFACTORY COLLOID CYST Michael Doukas1, George Alexiou2, Andreas Zygouris2, Dimitrios Pahatouridis2, Spyridon Tsiouris3, Sevasti Kamina1, Andreas Fotopoulos3, Spyridon Voulgaris2, Maria Bai1, Ann Goussia1 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Neurosurgery, Medical School, University of Ioannina, Greece 3 Department of Nuclear Medicine, Medical School, University of Ioannina, Greece Background. Colloid cysts are rare benign intracerebral lesions accounting for 0.2% to 2% of all intracranial neoplasms. They are predominately located in the anterior aspect of the third ventricle; however, there are occasional reports of colloid cysts found in the fourth ventricle, cerebellum, leptomeninges, parietal region and frontal lobe. We present herein a highly unusual case of a colloid cyst residing in the olfactory lobe. Materials and Method. A 74-year-old patient underwent a brain CT scan that incidentally revealed a space-occupying lesion measuring approximately 2.5X1.8X2.5 cm, involving the olfactory lobe. Magnetic resonance (MR) imaging showed the lesion as hyperintense on T1, T2 and FLAIR sequences, without surrounding brain edema. The differential diagnosis included a mucocele, dermoid cyst, meningioma and a glioma. Brain scintitomography showed a lesion of low metabolism, a finding consistent with benignity. The patient underwent surgery and intraoperative a cystic lesion was revealed. The cyst was eroding the dura and the frontal lobe and due to the danger of CFS leak, a dura repair was performed Results. Histological examination of the resected material showed that the cyst was lined by a single and focally by a pseudostratified layer of columnar ciliated epithelial cells resting

on a basal lamina. Mucus-filled cells were not found. The content of the cyst was consisted of an amorphous, eosinophilic, colloid-like material. Immunohistochemical examination showed positive staining of the lining cells for cytokeratin, epithelial membrane antigen and focally for carcinoembryonic antigen. The histologic diagnosis was consistent of a colloid cyst. Conclusion. Colloid cysts should be included in the differential diagnosis of lesions in the anterior fossa and that although benign they may be aggressive by eroding the dura and producing a mass effect. PP4-206 SEVERE PRIMARY INTRACEREBRAL HAEMATOMA – CORRELATIONS BETWEEN MAIN CLINICAL, IMAGISTIC AND MORPHOLOGICAL ASPECTS Iancu Emil Plesea1, Stelian Danut Enache2, Corneliu Cristian Georgescu1, Dan Cioroianu1, Mihai Popescu1, Simona Bondari1, Oltin Tiberiu Pop1, Alexandru Camenita2, Cornelia Enache2 1 University of Medicine and Pharmacy Craiova, Romania 2 Emergency County Hospital Craiova, Romania Background: The study is an integrated assessment of clinical, imagistic and morphological parameters in patients with severe primary intracerebral hemorrhage (PICH) who died during hospitalisation. Method. The selected group consisted of 183 cases and was divided into 2 groups: 102 cases who lived more than 48 h and were confirmend by computed tomography (CT) and 82 cases that died within 48 h and were confirmed by authopsy. The study was retrospective. The studied material consisted of patient’s medical records (medical records, CT films, autopsy protocols and histopathology - HP records). Assessed parameters were: clinical (seasons relation, age, sex, arterial blood pressure - HT, motor deficit - MD, degree of coma - C, Glasgow score at admission) and morphological static (PICH sites, size, perilesional edema - pE, microhaemorrhages - mH,) and dynamic (mass effect - ME, ventricular effusion - VE and subarachnoid effusion - SE). Results: The severe PICH showed a predilection for winter and summer but men (M) are affected more frequently in transition seasons. Sex distribution showed a slight M predominence. The most affected life decades are the 5th and the 7th with a regressive trend for M after the 6th decade and an increasing trend for women (W) towards the 7th decade. Around 70% of the patients had IIIrd and IVth stage HT at admission, 85% had MD and almost 50% Glasgow scores lower than 6. The most common sites of PICH were cerebral hemispheres, slightly more frequent in the left one. They had huge dimensions as compared to hosting encephalic structures. In lobar sites, more than one lobe were involved, mostly parietal, temporal and frontal lobes. Both CT and HP examinations revealed in many cases multiple mH both near and distant to PICH. Other morphologic predictive factors as pE and ME were constantly present. The VE involved at least one of the lateral ventricles. C is more frequent in W, after 60 years of age, in PICH occuring in winter or summer, and non-lobar PICH. That is also the case of MD, which is more frequent in PICH occuring in autumn and winter and lobar PICH. VE is more frequent in M, after 60 years of age, in PICH occuring in autumn and lobar PICH. Although less frequent, SE is slightly more frequent in W, PICH occuring in winter and non-lobar PICH, regardless of age. Conclusions: Our data suggest that PICH associated with one or more clinical and morphological poor outcome predictors (HT, MD, C, ME, VE and SE) results in patient's death, despite any sustained therapeutical intervention.

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PP4-207 ANAPLASTIC MENINGIOMA ASSOCIATED WITH MENINGIOANGIOMATOSIS: CASE REPORT OF A RARE ENTITY Suzan Zorludemir1, Seyda Erdogan1, Naciye Ozeren1, Kenan Bicakci2, Ilhan Tuncer1 1 Cukurova University, Medical Faculty, Department of Pathology, Turkey 2 Cukurova University, Medical Faculty, Department of Radiology, Turkey Background: Meningoangiomatosis (MA) is a rare malformative lesion that is associated with seizure and/or headache. It occurs both in patients with neurofibromatosis (NF), as well as in sporadic cases. Rarely, MA has been described coexisting with meningiomas. As far as we could reach, anaplastic meningioma together with MA has not been reported yet. Material and Methods: A 16-year-old boy had 2 year history of seizure. There was no history of NF. Magnetic Resonance Imaging (MRI) at the second year revealed a contrast-enhancing lesion, with dural tail and peritumoral edema near the left tentorium.The lesion was completely excised. Results: Microscopically, the lesion composed of mitotically active spindle cells with moderately large hyperchromatic nuclei, in a coarsely fasicular pattern resembling sarcoma. Limited areas showed elongated cells in interwoven fasicles and in whorls with psammoma bodies consistent with transitional meningioma. In the cortex, there were abundant small intracortical vessels ensheathed by meningothelial cells; typical of meningoangiomatosis. Conclusion: We present a unique case of anaplastic meningioma which have arisen from meningoangiomatosis without NF-2 history and discuss the pathogenesis, radiologic, clinic and pathologic features on the base of the literature. PP4-208 ANGIOCENTRIC GLIOMA: A RARE EPILEPSY-ASSOCIATED NEOPLASM Seyda Erdogan1, Suzan Zorludemir1, Tarık Tihan2, Filiz Cevlik1, Metin Tuna3, Erol Akgul4 1 Cukurova University, Medical Faculty, Department of Pathology, Turkey 2 University of California, San Francisco (UCSF), Department of Pathology, USA 3 Cukurova University, Medical Faculty, Department of Neurosurgery, Turkey 4 Cukurova University, Medical Faculty, Department of Radiology, Turkey Background: Angiocentric glioma is a distinct epilepsy-related entity which was not included in the World Health Organization (WHO) classification of Central Nervous System (CNS) Tumors yet. There are only two reports with 18 cases in the literature. Material and Method: A 3-year-old girl was admitted to the hospital with a history of seizure. MRI showed lesion at the right hippocampus and excised. Result: Microscopically, the lesion was composed of cells that were strikingly angiocentric to both large and small vessels. Tumor cells were uniform and usually bipolar. Atypical mitosis, necrosis were absent. Proliferation index was also negative with Ki-67. Conclusion: We present a case of newly described entity which should take part in the WHO classification of CNS tumors. And discuss the clinic, radiologic, histologic and ultrastructural features of this tumor on the base of the literature.

PP4-209 HISTOLOGICAL DISCREPANCIES IN MALIGNANT GLIONEURONAL TUMORS: A REPORT OF 4 CASES Aydın sisag1, Nalan Nese1, Eren Demirtas2, Mine Tunakan3, Turkan Rezanko3, Cuneyt Temiz4, Hasan Mirzai4 1 Celal Bayar University, Faculty of Medicine, Department of Pathology, Manisa, Turkey 2 Mikro Pathology Laboratory, zmir, Turkey

3 Ataturk Training and Research Hospital, zmir, Turkey 4 Celal Bayar University, Faculty of Medicine, Department of Neurosurgery, Manisa, Turkey Background: Although not included in WHO 2000 Classification of CNS tumors, malignant glioneuronal tumor (MGNT) is widely accepted as a new entity which resemble any type of high grade (grade III or IV) malignant glioma with tumor cells expressing both neuronal and glial markers without mature ganglion-like cells. In this case series, 4 immunohistochemically proven cases of MGNT are reported in order to emphasize the histological discrepancies among seperate cases of this poorly understood tumor group. Cases: Four cases of MGNT from three different institutions are studied. Two of the patients were female and the other two were male, age 13 (Case 1), 50 (Case 2), 70 (Case 3) and 18 (Case 4) years and localizations were right parietooccipital-intraventricular, left occipital, right temporal and right occipital, respectively. All cases showed peritumoral oedema and contrast enhancement. Except for Case 1, tumors were well circumscribed and gross-totally excised followed by radio-chemotherapy. Patients Case 1 and Case 3 died because of the disease 12 months and 21 months after the initial diagnosis with recurrences while others are still alive for 16 months (Case 2) and 5 months (Case 4). For all cases, glial fibrillary acidic protein, synaptophysine, neurofilament protein and Ki-67 were applied and it was shown that first three of these markers were much or less co-expressed by the tumor cells despite the histological discrepancies among cases. Case 1 and Case 3 was composed of PNET-like areas and glioblastoma areas. Case 1 additionally contained loose and spindle-cell areas while Case 3 showed perivascular arrangement of tumor cells and some pseudopapillary structures. The bulk of the tumor in Case 3 was composed of areas of typically oligodendroglioma or anaplastic oligodendroglioma, which are found in also Case 4. Case 4 also presented psedopapillary structures, spindle-cell areas and epitheliod gemistocyte-like cell clusters intermingled with indifferentiated small cells. Necrosis, pseudopalisading of tumor cells around necrosis and brisk mitotic activity were apparent in all cases. Ki-67 proliferation indices were 55%, 72%, 20% and 43%, respectively. Conclusion: As histology of MGNTs can diverge greatly, all CNS tumors resembling malignant gliomas (especially glioblastoma) or supratentorial PNET should undergo immunohistochemical analyses with at least two neuronal markers as well as with glial fibrillary asidic protein in order to make accurate diagnosis and prognostic presumptions. PP4-210 SYMPTOMATIC SPINAL LUMBAR JUXTAFACET CYST: A CASE REPORT Evangeli Lampri1, George Alexiou2, Michael Doukas1, Eufemia Balasi1, George Fotakopoulos2, Spyridon Voulgaris2, Dimitrios Stefanou1, Ann Goussia1 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Neurosurgery, Medical School, University of Ioannina, Greece Background: Juxtafacet cysts (JFC) are rare intraspinal lesions associated with the facet joints. Although they can be seen along the spinal cord, the lumbar region is the most commonly affected area. The putative mechanisms of cysts formation include spinal instability, arthropathy and trauma. Histologically, two types of JFC are recognized: the synovial and ganglion cysts or

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pseudocysts. The two lesions have different histological features, however, studies based on detailed histological analysis revealed dual findings in the same sample, suggesting that these cysts are either parts or progression of the same disease. We report herein a case of symptomatic juxtafacet cyst presenting as L4-L5 radiculopathy. Case report: A 53-year-old man presented with a history of progressively increasing severe pain in the right lower extremity. Physical examination revealed a L4-L5 sensitive and motor deficit. Magnetic resonance imaging (MRI) revealed an intraspinal cystic mass at the segment L4-L5, with hypointense T1-weighted and hyperintense T2-weighted signal and peripheral enhancement. Surgical treatment was planned due to severe pain and neurologic deficit. Histological examination of the resected material showed the presence of a cystic formation with no evidence of epithelial or synovial lining. The wall of the cyst was fibrovascular, with myxoid areas, focal hemorrhage, rare hemosiderin deposits, collections of multinuclear giant cells and small calcifications. The immunohistochemical staning for glial fibrillary acid protein (GFAP) and neurofilament (NF) were negative, showing that the lesion was not derived from neural tissue. The diagnosis was consistent with a cystic formation of spine or juxtafacet cyst with morphology of a ganglion cyst. The postoperative course was uneventful and the patient was discharged on the third following surgery day. On follow-up examinations at 3 months and 1 year after surgery, the patient remained free of pain and symptoms. Conclusion: Juxtafacet cysts are uncommon cause of radiculopathy. We emphasize the need to be thoroughly familiar with this pathologic entity that should be considered in the differential diagnosis of spinal lesions. PP4-211 EMBRYONAL TUMOR WITH ABUNDANT NEUROPIL AND TRUE ROSETTES: A RARE PEDIATRIC EMBRYONAL NEOPLASM OF CNS. REPORT OF FOUR CASES Marco Gessi1, Marina Gardiman2, Concezio Di Rocco3, Riccardo Riccardi4, Palma Maurizi4, Libero Lauriola5, Felice Giangaspero6 1 Div. of Neuropathology, National Neurological Institute “C. Besta”, Milan, Italy 2 Dept. of Pathology, University of Padova, Padova, Italy 3 Div. of Pediatric Neurosurgery, Catholic University, Rome, Italy 4 Div. of Pediatric Oncology, Catholic University, Rome , Italy 5 Dept. of Pathology, Catholic University, Rome, Italy 6 Dept. of Experimental Medicine and Pathology, University of Rome “La Sapienza”; IRCCS INM Neuromed, Pozzilli (IS), Italy BACKGROUND Embryonal neoplasms of CNS affect early years of life and share an aggressive behavior. WHO classification includes five different well-defined entities: medulloepithelioma, ependymoblastoma, medulloblastoma, supratentorial PNET and ATRT. Recently, a new tumor combining features of ependymoblastoma and neuroblastoma named “embryonal tumor with abundant neuropil and true rosettes” have been reported. This lesion is characterized by the presence of areas of well-differentiated neuropil containing ependimoblastic rosette and undifferentiated neuroepithelial cells resembling classic PNET. We report four cases of this rare entity. METHOD The tumoral specimens were routinely processed, paraffin embedded, and H&E stained. Slides were processed for immunohistochemistry using antibodies for Vimentin, S-100, Desmin, Synaptophysin, Neurofilaments, Smooth Muscle Actin, GFAP and BAF47/INI1. The proliferation index was evaluated using anti-MIB-1 antibody. RESULTS Patient #1 was a 1.5 years-old female with a right frontal lesion; patient #2 was a 1.5 years-old male with fronto-temporal lesion; patient #3 was a 3 years–old female with bi-frontal tumor; patient #4 was a 1.5 with pontine lesion. The tumors showed a variable cellular density with fields composed by fibrillar neuropil-like matrix and by

hypercellular areas, formed by small hyperchromic cells. Rare Homer-Wright rosettes were focally observed. Mitoses and apoptosis were frequent. The distinctive feature was the presence of numerous ependimoblastic rosettes, found in the hypercellular areas and/or in the fibrillary neuropil-like matrix. The rosettes were composed by pseudo-stratified embryonal cells, arranged around a central lumen. In one case, a sarcomatous-like component of elongated pleomorphic cells was present. The undifferentiated cells were negative for GFAP and synaptophysin whereas the neuropil-like matrix stained intensely for sinaptophysin and neurofilaments. The sarcomatous areas present in one case showed diffuse immunoreactivity for smooth muscle actin and for desmin. The tumors showed a nuclear staining for BAF47/INI1. The MIB-1 ranged from 10 to 30 % in the higher cellular areas. Interestingly, the ependimoblastic rosettes in the neuropil-like areas contained the more proliferative cellular component. CONCLUSION Embryonal tumor with abundant neuropil and true rosettes although presents some features common to other embryonal tumors, seems to have specific histopathological, neuroradiological and clinical features, delineating it as a new entity among pediatric brain tumors. PP4-212 EXPRESSION OF NUCLEAR FACTOR – B IN HUMAN ASTROCYTOMAS: RELATIONSHIP WITH PI BA AND PROGNOSTIC SIGNIFICANCE. Georgia Levidou, Angelica A Saetta, Penelope Korkolopoulou, Elias El-Haber, Polychronis Demenagas, Irene Thymara, Kalliopi Diamantopoulou, Efstathios Boviatsis, Euphtemia Thomas-Tsagli, Ioannis Panagiotidis, Efstratios Patsouris National and Kapodistrian University of Athens, Department of Pathology, Athens, Greece Background: NF- B is a family of dimeric transcription factors that play a critical role in host defence by regulating the expression of immune genes. Several investigators have reported its constitutive activation in various human tumour cell lines, whereas its immunoexpression has been regarded as a marker of poor prognosis in several tumours. NF- B activation is tightly regulated by its interaction with I B inhibitory proteins. However, little information is available about the role of I B phosphorylation in relation to NF B activation, and the clinical significance of this interaction in astrocytomas. This study aimed to elucidate the role of NF B in astrocytomas, focusing on p50/ F B1 subunit, to determine its association with phosphorylated I B, and to define the clinical impact of this interaction. Method: The levels of p50/NF B1 and pI Ba proteins expression were quantified immunohistochemically in paraffin-embedded tissue from 73 patients with astrocytomas, by using a rabbit polyclonal anti-NF B p50 and a mouse monoclonal anti-pI Ba antibody. Nuclear and cytoplasmic immunoreactivity was assessed separately for both antibodies. Results: Nuclear NF B1/p50 immunoreactivity was detected in 66/72 cases (91.6%), whereas cytoplasmic immunoreactivity in 17/72 (23%). Concurrent cytoplasmic and nuclear expression was recorded in 17 cases (22%). pI Ba expression was found in 72/73 (98.6%) cases and was predominantly cytoplasmic with scattered nuclear positivity. Concurrent pI Ba and nuclear NF B1/p50 immunoexpression was detected in 66 cases. All cases with nuclear NF B1/p50 expression manifested pI Ba immunoexpression. pI Ba expression was positively correlated with nuclear NF B1/p50 expression (p=0.0054) and adversely correlated with cytoplasmic NF B1/p50 expression (p<0.0001). Nuclear NF B1/p50 and pI Ba expression increased in parallel with tumour grade (p= 0.0031 and p<0.0001). In univariate survival analysis nuclear NF B1/p50 and pI Ba expression adversely affected survival (p<0.0001 and p=0.002). Multivariate survival analysis selected NF B1/p50 nuclear expression (p=0.001) as a significant prognostic factor. Conclusion: Our study reinforces the concept that NF B1/p50 activation is largely dictated by its interaction with I Ba in human astrocytomas.

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Additionally, NF B1/p50 nuclear expression parallels tumour grade, a result that suggests its contribution to tumour agressiveness. Finally, our results suggest that nuclear NF B1/p50 expression may serve as a useful independent marker for stratifying patients with astrocytoma in terms of prognosis. PP4-213 CEREBRAL NEUROBLASTOMA IN ADULTS: REPORT OF THREE CASES Marco Gessi1, Michele Bisceglia2, Libero Lauriola3, Felice Giangaspero4 1 Div. of Neuropathology, National Neurological Institute “C. Besta”, Milan, Italy 2 Div. of Anatomic Pathology, Ospedale “Casa Sollievo della Sofferenza”, S. Giovanni Rotondo, Italy 3 Dept. of Pathology, Catholic University, Rome, Italy 4 Dept. of Experimental Medicine and Pathology, University of Rome “La Sapienza”; IRCCS INM Neuromed, Pozzilli (IS), Italy BACKGROUND: The term “cerebral neuroblastoma” identifies an embryonal neoplasm of CNS belonging to the group of the supratentorial PNET (sPNET) showing various degree of neuronal differentiation. These tumors affect preferentially infants or children and appear to be extremely rare in adult life. We present three cases of cerebral neuroblastoma in adult patients. METHOD: Tumoral specimens were routinely processed, paraffin embedded, and H&E stained. Slides were processed for immunohistochemistry with the ABC system, using antibodies for Vimentin, S-100, Desmin, Synaptophysin, Chromogranin, Neurofilaments, Smooth Muscle Actin, EMA, CK, MAP-2, GFAP, p53 and Neu-N. The proliferation index was evaluated with anti-MIB-1 antibody. RESULTS: In patient 1, a 63 years-old female, MRI revealed a 4 cm infiltrating lesion affecting the left occipital lobe, with non-homogeneous contrast enhancement and necrosis. In patient 2, a 61 years old female, MRI showed a 7 cm well-demarcated right frontal mass, with intense non-homogeneous contrast enhancement. In patient 3, a 59 years old female, CT scans revealed a left parieto-occipital lesion with edema, midline shift and homogeneous contrast enhancement. Histopathological examination showed lesions with similar architecture but with large spectrum in cell composition. Tumors were formed by lobules separated by thin fibro-vascular septa, filled by a mixture of small elements with neuroblastic features as well as larger cells with neuronal or ganglion-like morphology, merged in a variable amount of neuropil. However, cells showed also a variable range of “anaplasia” characterized by bizarre giant cells, apoptotic bodies, necrosis and atypical mitoses. Immunohistochemistry revealed a variable positivity for synaptophysin, neurofilaments, chromogranin and Neu-N in isolated cells. GFAP and mesenchymal or epithelial markers were negative. The proliferation index, assessed by Ki67/MIB1 antibody was variable but focally elevated. p53 was negative. CONCLUSION: Cerebral neuroblastomas in adults are histologically similar to those occurring in pediatric age, although they seem to show a higher degree of severe anaplasia. They lack specific clinico-radiological features. Moreover they have to be distinct from malignant neoplasms more frequent in the adult age group as mixed glio-neuronal tumors and pure gliomas such as glioblastoma and anaplastic oligodendrogliomas. PP4-214 ANCIENT SCHWANNOMA OF THE SPINAL CORD. A RARE CASE Maria Papaevangelou, Efthymios Koniaris, Eleni Psichogiou, Maria Gazalidou, Amalia Patereli, Maria Sevastiadou, Nikiforos Apostolikas Pathology Department Anticancer Oncological Hospital of Athens, ‘St. Savvas’, Athens, Greece

Background: Schwannomas of the Spinal Cord (SSC) are rare benign tumors. Most are sporadic and some can be part of hereditary syndromes. Histological malignancy of this neoplasm is rare. Method: A 51 year old patient admitted to our hospital for neurologic evaluation with symptoms of spinal compression because of a CT scan lesion on the spinal cord. The lesion was surgically removed and we received a well-circumscribed, elongated, cylindrical specimen which was measuring 8.4x2.2cm. The tumor was encapsulated and on dissection it was partly solid with whitish tint and partly multicystic. The cysts were filled with a bloody-serous liquid. Results: Histologically, multiple cysts comprised most of the tumor while the rest consisted of an inconspicuous or focally cellular component (Antoni type A) with a hint of nuclear pallisading that rendered the diagnosis difficult. Hyalinization, hemosiderin deposition and some nuclear atypia were the minor characteristics of the tumor. In Antoni B areas edema separated tumor cells. No mitotic figures were found. Immunohistochemically the tumor showed reactivity for S-100 protein, vimentin and CD68. Keratin, desmoplakin, neurofilaments and desmin are not expressed. Conclusion: In this presentation we aim to emphasize the polycystic appearance of a so called ancient SSC in a rare location. SSCs generally agreed that are neoplasms originating from Schwann cells, hence their name. Malignant transformation is an exceptionally rare event. Ancient SSC are usually large tumors of long duration. Although histologically are benign, biologically mimick malignancy. PP4-215 CONGENITAL SUPRATENTORIAL CYSTIC HEMANGIOBLASTOMA; A CASE REPORT AND REVIEW OF THE LITERATURE Hakan Karabagli1, Pınar Karabagli2 1 Konya Numune Hospital, Neurosurgery Clinic, Turkey 2 Konya Education and Research Hospital, Patology Laboratory, Turkey Congenital supratentorial hemangioblastomas are rarely encountered tumors even in pediatric population. Extensive review of the literature revealed that approximately 118 cases have been reported in the literature so far. However, only 6 of these occurred in infants, and 4 of them occurred during the first two months of life. A 5-week-old boy presented with emesis, irritability, bulging anterior fontanelle and a gradually expanding head circumference since birth. His previous medical and family history was uninformative in terms of cancer or inherited diseases. The MR scans demonstrated a large loculated cyst with a 3-cm heterogeneus mural nodule, first pushing the left frontal and parietal lobes, and then displacing in this region. The histopathologic diagnosis was reticular variant of hemangioblastoma. We reviewed the literature on congenital supratentorial hemangioblastoma and discussed the histopathologic characteristics and differential diagnosis associated with such lesions in this report PP4-216 THE EXPRESSION OF OSTEOPONTIN IN HUMAN ASTROCYTOMAS: INTERSTITIAL EXPRESSION CORRELATES WITH MALIGNANCY GRADE AND ANGIOGENESIS Koviljka Matušan1, Senija Behrem1, Nives Jonji 1, Kamelija Žarkovi 2, Ksenija Lu in1 1 Department of Pathology, Rijeka University School of Medicine, Rijeka, Croatia 2 Department of Pathology, Clinical Hospital Rebro, Zagreb, Croatia BACKGROUND: Osteopontin (OPN) is a phosphorylated protein secreted into the extracellular matrix by a variety of cell types. Numerous functions have been ascribed to osteopontin, including roles in bone remodelling, cell-adhesion, cell-mediated immunity, the ability to act as a cytokine in cell signalling,

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resulting in proliferation and/or cell survival, and angiogenesis. OPN has also been detected in a number of human tumour tissues and assessed as a potential marker of tumour progression. METHOD: This study was aimed to analyse by immunohistochemistry the expression of OPN in 76 human gliomas of various grades of malignancy: 8 pylocytic astrocytomas, 10 grade 2 astrocytomas, 8 anaplastic astrocytomas and 50 glioblastomas. The staining results were scored in a semi-quantitative manner by assessing the percentage of positive tumour cells and the intensity of staining. We also assessed the intercellular distribution of staining, which was categorized as focal or diffuse, with the staining intensity expressed either as low or high. Since OPN has been implicated in angiogenesis, we analysed the correlation between OPN expression and microvessel densitiy in glioblastomas. Microvessels were stained with -endoglin antibody (CD105) and counted in predominantly vascular areas (hot spots) at x 400 magnification. RESULTS: In normal brain tissue the expression of OPN was present in some glial and neuronal cells in the form of cytoplasmyc granular staining of low intensity, while microglial cells were strongly positive. In glial tumors intracellular OPN expression ranged from absent or low (score 0-5) to strong (score 6-10). Strong OPN expression was observed in 3 grade I (37.5%), 4 grade II (40%), 2 grade III (25%) and 21 grade IV (42%) astrocytomas. In some tumors OPN was also present interstitially, between tumor cells. This staining pattern was accentuated around tumor necrosis and, in most of the cases, also around blood vessels. Tumours with either a diffuse or strong expression were considered positive in intercellular distribution of OPN staining. Interstitial OPN expression was observed in 1 pylocytic astrocytoma (12.5%), 2 grade II astrocytomas (20%), 2 anaplastic astrocytomas (25%) and 25 glioblastomas (50%). We found the association of interstitial OPN expression and angiogenesis i.e. microvessel density was higher in tumors with high interstitial OPN expression (Mann Whitney test, p<0.03). CONCLUSION: Our results show the overexpression of OPN in human astrocytic tumors and suggest the role of OPN in astrocytoma progression and angiogenesis. PP4-217 CAFFEIC PHENETHYL ESTER (CAPE) SUCESSFULLY REDUCES BRAIN DYSFUNCTION IN SEPSIS Huseyin Fidan1, Onder Sahin2, Yuksel Ela1, Aynur Kilbas3, Olcay Eser2, Murat Cosar2, Irfan Altuntas3 1 Department of Anesthesiology, Afyon Kocatepe University School of Medicine, Afyonkarahisar, Turkey 2 Department of Pathology, Afyon Kocatepe University School of Medicine, Afyonkarahisar, Turkey 3 Department of Biochemistry, Suleyman demirel University, School of Medicine, Isparta, Turkey Background and goal of study: Sepsis and ensuing multi organ failure continue to be the major causes of morbidity and mortality in the intensive care units. Nuclear factor-Kappa beta (NFKB) activation is supposed to be one of the targets in the treatment of sepsis and ensuing mortality. We studied the effectiveness of caffeic phenethyl ester (CAPE), a known NFKB inhibitor, in cecal ligation and puncture (CLP) induced sepsis model of brain dysfunction. Materials and Methods: 40 rats are randomized to 5 groups. All rats were operated to induce sepsis with cecal ligation and puncture (CLP) except control and CAPE groups that were operated just with laparotomy. CAPE (50 μmole/kg) was administered to rats intramuscularly at the time of operation in CAPE and CA+SEP (0) groups. CAPE was administered to rats in CA+SEP (12) group 12 hours after CLP. All rats from each group were sacrificed 24 hours after CLP; half of brain coronally was taken out for histopathological, and the rest brain was taken out for oxidative stress parameters. Apoptosis was examined with Tunnel staining. Induced nitric oxide synthase (iNOS) and heat schock protein(HSP70) were examined with immunohistochemistry. Malonlydialdehyde (MDA), catalase

(CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were studied for oxidative stress evaluation. Results and Discussion: Immunohystochemical studies revealed that sepsis induced brain dysfunction in our model. Histopathologically apoptosis and iNOS and HSP70 expressions were studied in hippocampal CA1 region, prefrontal cortex and plexus choroideus. Although apoptotic measurement revealed no change between groups in hipocampal CA1 region, iNOS and HSP70 expressions were significantly increased in all other groups in comparison to control group. However, iNOS and HSP70 expressions were significantly reduced as CA+SEP (0) < PP4-219 MEDICALLY INTRACTABLE EPILEPSY: REPORT OF THREE CASES Ozgur Mete1, Bilge Bilgic1, Cicek Bayindir1, Murat Imer2 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey 2 Istanbul University, Istanbul Faculty of Medicine, Department of Neurosurgery, Istanbul, Turkey Background: There is no precise definition of intractable epilepsy. Among the considerations are seizure frequency, seizure type, severity of attacks, and impact on quality of life. Twenty percent of epilepsy is intractable and surgery is an option in the attempt to cure or reduce the severity of medically resistant cases. We report herein 3 intractable epilepsy cases diagnosed as cortical dysplasia, Rasmussen encephalitis and Sturge-Weber syndrome for discussing their histopathological features. Case 1: A 2-year-old male has been suffering from generalized tonic and clonic seizures since 15 months. The histopathological examination revealed cortical dislamination, neuronal heterotopia in the white matter. Neuroimaging and histopathological findings were found compatible with cortical dysplasia. Case 2: A 14-year-old male has been suffering from progressive, focal and generalized seizures and also left hemiparesy on the forearm and leg since 3 years. Cranial MR showed right frontal and temporal atrophy. Functional right hemispherectomy was performed. The histopathological findings revealed neuronal loss, neuronophagia, microglial nodules and perivascular lymphocytic infiltration. The clinical, radiological and histological findings let us to diagnose the case as Rasmussen encephalitis. Case 3: A 2-year-old male has been suffering from progressive, generalized and focal seizures since 20 months. A pink macular lesion in the dermatome of N.trigeminus was remarked. MRI showed cortical atrophy and also increase in the thickness of leptomeninges. Being accepted medically intractable epilepsy, surgical procedure was performed. Leptomeningeal angiomatosis and cortical atrophy let us to diagnose Sturge-Weber syndrome with the presence of port-wine stains. Conclusion: Surgery in epilepsy is the resection of epileptogenic focus The pathological lesions are classified recently as: (a) hippocampal sclerosis (b) malformative lesions, (c) vascular lesions (d) tumors, (e) chronic inflammatory and infectious lesions, (f) ulegyric lesions (g) gliosis-astrocytic proliferation, (h) concomitant calcified lesions and (i) unspecified lesions. It is impotant to remember that Rasmussen encephalitis is a rare, chronic inflammatory disease that usually affects only one hemisphere. Furthermore, special attention has been given to the coexistence of focal cortical dysplasia and tumors, which include mixed neuroglial tumors. Finally, our cases and literature data stress that clinical, neuroradiological and histopathological correlation is important for the diagnosis of these lesions. PP4-220 PROGNOSTIC FACTORS IN MENINGIOMAS Deniz Ozcan1, Aziz Hatiboglu2, Celal plikcioglu2 1 Okmeydani Training Hospital, Pathology Department, stanbul, Turkey

2 Okmeydani Training Hospital, Neurosurgery Department, stanbul, Turkey

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ABSTRACT: Meningiomas are mostly benign tumors cured by surgical resection. But the behaviour of meningiomas are sometimes diffucult to predict. Some of them tend to recur, therefore longterm management in subtotally resected tumors remain controversial. The microscopic brain invasion, mitotic rate higher than 4/ 10 HPF, the presence sf sheeting, hipercellularity, prominent nucleoli, nuclear pleomorfism, small cells, decreased recurrence free survival (RFS). In this study MIB-1 LI , PR, ER, P 53 and EGFR expressions are compared with histopathological changes. METHOD: 148 meningioma cases operated in our Neurosurgical Department of Okmeydanı Hospital 20 cases were chosen for study.( 4 angiomatous meningioma, 2 of which recurred, 5 meningioma cases, 5 atypical grade II meningioma cases, 6 malignant meningioma cases) are examined immunohistochemically using MIB-1 monoclonal antibody estrogen-progesteron receptor P53 and EGFR. RESULTS: There were a close association between MIB-1 LI indices and mitotic index and tumor grade. According to Spearman’s rho ( Spearman’s Rho= 0,493 ) ( p=0,027 ) P53 and EGFR immunohistochemistry revealed no useful prognostic information. ER, PR immunohistochemistry were variable, but usually ER were not expressed. PP4-221 EXPRESSION OF HORMONE RECEPTORS PR & ER, ANGIOGENIC FACTORS VEGF – A & VEGF-R3, P53, MDM2, CERBB2 AND CD117 IN MENINGIOMAS Kalliopi Diamantopoulou1, Euphemia Thoma-Tsagli1, Penelopi Korkolopoulou2, Vassilia Leodara1, Ioannis Antoniadis3, Konstantinos Kouzelis3 1 Pathology Department Gen Hospital Asklepieio Voula, Greece 2 Pathology Department University of Athens, Greece 3 Neurosurgery Department. Gen. Hospital Asklepieio Voula, Greece Background: Meningiomas are mostly frequent and benign central nervous system tumors. Independently from the extent of tumor resection, reccurence and tumor progression often determine clinical outcome. In an attempt to understand meningioma tumor biology, we evaluated biomarkers possibly involved in tumor formation, reccurence and malignant transformation, such as progesterone and estrogen receptors, angiogenic factors VEGF – A and VEGF-R3, p53, MDM2, cerbB2 (Her-2/neu) and CD117 (c-kit) and their possible implication in therapeutic approaches. Methods: We examined 34 meningiomas operated in our hospital the last 4 years, 30 of which benign, 2 reccurent (WHO grade I) and 2 atypical (WHO grade II) immunohistochemically for Progesterone Receptors (PR), Estrogen Receptors (ER), Vascular Endothelial Growth Factor (VEGF-A) and its Receptor (VEGF-R3), p53, MDM2, cerbB2 and CD117. Results: PR were expressed in 27/32 (84%) WHO grade I meningiomas in a mean of 44,15%. ER were demonstrated in 12/32 (37,5%) in a mean of 5,5%. Neither PR nor ER were expressed in atypical meningiomas. VEGF-A was intensly demonstrated in most WHO grade I and II meningiomas mainly with cytoplasmic granular or dot-like distribution. In 2 reccurent cases it was faintly or negatively expressed. VEGF-R3 was variably detected with cytoplasmic and nuclear distribution in all meningiomas. p53 nuclear staining was shown in 18/32 (56,25%) benign meningiomas in a mean of 13,05% with negative staining in 2 atypical meningiomas. Nuclear MDM-2 overexpression was variably demonstrated in all benign, recurrent and atypical meningiomas in a mean of 60,90%. cerbB2 was scored 3+ positive staining in 1 benign meningioma (2%), 2+ in 6 meningiomas (17,6%), 1 of which atypical and 1+ and less in the rest (79,4%). CD117 was expressed focally and faintly in 4/34 (11,7%) benign mengiomas. Conclusion: Loss of progesterone receptors’ expression is shown to be associated with increased reccurence, implying a better prognosis for positive expression, as were most of our benign and 2 atypical cases. Instead, presence of ERs in meningiomas may correlate with karyotype

abnormalities. Hormone receptor status, especially in females, seems to be a possible anti-hormonal target of therapy. Angiogenic factors like VEGF-A and its Receptor VEGF-R3 may be involved in meningioma vasculature and likelihood of reccurence, so antiangiogenic agents should be encountered in therapy. Inactivation of p53 tumor suppressor and overexpression of MDM2 protein may promote meningioma progression and future reccurence, revealing a possible candidate to an aggressive treatment. In our cases the amount of expression of MDM-2 seems not to be correlated to histological grade and reccurence. cerbB2 (Her-2/Neu) and CD117 (c-kit), both members of the tyrosine kinase receptors’ family, have not yet been detected in meningiomas but their possible overexpression, especially cerbB2, as in our limited number of cases, must be studied further. PP4-222 TYPICAL AND UNUSUAL PATHOLOGICAL CHARACTERISTICS AND RADIOLOGICAL FINDINGS IN PRIMARY NEOPLASMS OF THE SPINAL CORD: UCSF EXPERIENCE Agne Naujokas, Ashley Aiken, Philip Weinstein, Cynthia Chin, Tarik Tihan UCSF, USA Background: Limited biopsy samples and overlapping clinical/radiological features can complicate the diagnosis of primary neoplasms of the spinal cord. Typical features of each entity are well known, but deviations from these features are not well recognized. Our study is aimed to determine typical as well as unusual radiological and pathological characteristics of the primary spinal cord tumors. Material and Methods: We reviewed the clinical, radiological and pathological features of all primary neoplasms of the spinal cord diagnosed and treated at our institution between 1996 and 2005. Radiological and pathological materials were analyzed using standard evaluation forms, and the results were analyzed using a statistical software package (SPSS 11.1 for Windows). Appropriate permissions were obtained from the institutional Committee on Human Research (H41995-25437). Results: The entire cohort of 136 patients with primary spinal cord neoplasms consisted of 79 males and 57 females with a mean age of 43 years (range 1-89 years). Only 13 (10%) of these neoplasms occurred in the pediatric age group (<15years). There were 57 ependymomas, 44 schwannomas, 15 neurofibromas, 10 astrocytomas of various grades, 8 hemangioblastomas, 2 gangliocytomas, and 1 subependymoma. Forty four percent of schwannomas demonstrated cystic change radiologically as well as pathologically, 32% had marked inflammatory infiltrates, and 48% had significant degenerative atypia. Radiological evidence of hemosiderin was most prevalent in ependymomas. A significant number of non-pilocytic tumors demonstrated Rosenthal fibers in the neuropil surrounding the tumor. Radiologically some ependymomas could not be distinguished from schwannomas or pilocytic astrocytomas. Clinical features, radiological presence of cyst and T1-T2 characteristics were not helpful discriminators. Pathological material was insufficient in three cases, in which radiological features were helpful. Conclusion: In our series, a significant number of primary spinal cord tumors did not show typical features radiologically, and to a lesser extent histologically. Radiologically, presence of hemosiderin correlated best with an ependymoma, but otherwise pilocytic astrocytomas could not be distinguished from ependymal tumors. The main challenge in the pathological diagnosis was the sample size. Recognition of prevalence of atypical features is critical in correct diagnosis primary spinal cord tumors.

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PP4-223 CEREBRAL MEDULLOEPITHELIOMA Faten Limaiem1, Amina Mekni1, Salma Bellil1, Ines Chelly1, Raoudha Doghri1, Slim Haouet1, Nidhameddine Kchir1, Mohamed Moncef Zitouna1, Mohamed Zemmal2, Khedija Bellil1 1 Department of Pathology, La Rabta Hospital, Tunisia 2 Department of Neurosurgery, La Rabta Hospital, Tunisia Introduction: Intracranial medulloepithelioma (ME) is an uncommon, highly malignant primitive neuroectodermal tumour of the central nervous system (CNS) which pathologically recapitulates the neural tube. It usually develops in early childhood and may occur anywhere within the CNS, the most common site being periventricular. Aim of study: The aim of our study was to describe clinicopathological features and immunohistochemical profile of ME and to discuss differential diagnosis. Patients & Methods: Between January 1991 and March 2007, 4 cases of cerebral medulloepithelioma were diagnosed at the pathology department of La Rabta Hospital. Medical record and microscopic slides were available in all cases and were retrospectively reviewed. Results: Our study included four male children aged between 4 and 13 years (mean age = 10,75 years). All patients presented with symptoms of increased intracranial pressure including headache vomiting and lethargy. CT scan showed an isodense or hypodense non-enhancing well-circumscribed tumour involving the temporoparietal lobe in two cases, the frontoparietal lobe in one case and the temporoparietooccipital lobe in one case. Gross-total resection of the tumour was performed in 3 cases and partial resection in one case. Pathological examination of the surgical specimen showed a tumoral proliferation made neural tube-like structures composed of primitive appearing cells that formed a pseudostratified epithelium arranged in tubular, papillary or trabecular configuration. The neural tube-like structures were outlined by a continuous, PAS-positive basement membrane. Mitotic figures were abundant and tended to be located near the luminal surface. Immunohistochemically, tumour cells were in all cases strongly positive for vimentin but were negative for PS100, NSE and GFAP. Postoperatively, 3 patients underwent adjuvant radiotherapy in the tumour bed. During the follow-up period which ranged between 1 and 3 years, one patient died. PP4-224 RT-PCR: DIAGNOSIS VALUE IN DOGS WITH SPONTANEOUS ACUTE-, SUBACUTE-, AND CHRONIC-DEMYELINATING DISTEMPER ENCEPHALITIS Edson M. Scarpelli1, Karime C. Scarpelli1, Alexandre M. Amude2, Geórgia F. Cintra1, Amauri A. Alfieri2, Maria L. Cintra1 1 Department of Pathology - UNICAMP (State University of Campinas) SP, Brazil 2 Laboratory of Animal Virology - UEL (State University of Londrina) PR, Brazil Background: Distemper is an endemic disease, that is not restricted to carnivorous or to a single specie. Encephalitis due to canine distemper virus (CDV) has been associated with multiple sclerosis (MS) because of extensive demyelization in both and the postulated viral origin in MS. A domestic dog barely spends its life without contact with distemper virus (DV), mainly in developing countries; also, contact may occur through vaccination programs. Methods: Within three years, 45 dogs with signs of canine distemper (CD) were attended in Zoonosis Control Center of Taubaté, São Paulo, Brazil. Clinical data were recorded according to a protocol. The animals were euthanized, necropsied, photos were taken, and a brain sample was randomly selected for RT-PCR to CDV. In 7 healthy dogs (control group), submitted to euthanasia for different reasons, the same procedure was adopted. This study followed guidelines prescribed by the Brazilian Medical Research Centre and received approval from the Research Ethics Committee of the State University of Campinas. Results: in 31 out 45 dogs, a positive RT-PCR

reaction was found in brain samples (group A); in the 14 remained dogs, negative results were obtained (group B); in all the 7 healthy dogs samples, positive PCR results were found (group C). On histological view, chronic-demyelinating encephalitis, typical of CD, was identified in group A animals and in 11/14 dogs in group B; 3/14 dogs in group B displayed no histological abnormalities. Focal lymphoid infiltrate was found in 5/7 animals and no abnormalities in 2/7 dogs in group C. Conclusion: For the endemic nature of CD and vaccination programs, positive results of RT-PCR do not allow the distinction of CDV from other diseases that share with it the same clinical manifestation. By the other side, false-negative RT-PCR could be the result of inadequate brain sample selection or be due to technical reasons. In this way, our findings stress the importance of clinical evaluation for proper CD diagnosis. The results may be useful for clinical research on CDV and sanitary programs of control of the disease. PP4-225 PRIMARY CENTRAL NERVOUS SYSTEM LYMPHOMA IN IMMUNOCOMPETENT PATIENTS: RETROSPECTIVE ANALYSIS OF 15 CASES Aylin Okcu Heper1, Ercan Armagan2, Ayse Ozgun1, Hasan Caglar Ugur2, Esra Erden1, Ali Savas3, Isinsu Kuzu1 1 Ankara University, School of Medicine, Department of Pathology, Ankara, Turkey 2 Ankara University, School of Medicine, Department of Neurosurgery, Ankara, Turkey 3 Ankara University, School of Medicine Department of Neurosurgery, Ankara, Turkey Background: Primary central nervous system lymphoma (PCNSL) is a rare disease. It represents approximately %3 of the central nervous tumors and is most often seen in inmmunodeficient patients. The incidence has increased in last two decades. In this study we aimed to review retrospectively PCNSL cases which were diagnosed in Ankara University, Faculty of Medicine, Department of Pathology with their clinical, responses to therapy regiments and survival. Material-method: Total 15 cases diagnosed in 2002-2006 and confirmed immunohistochemically were included into the study. The patients were analysed in relation to sex, age, time of the symptoms, procedures, treatments, survival and pathological features. Results: All of the patients (8 male, 7 female) were immunocompetant and the time of the initial symptoms to pathological diagnosis was in range between 1 month to 4 months. The mean age of the patients was 47.7 (range, 27-66 years). The most common symptom was hemiparesy (%80), headache (%46.6) and seizures (%26.6). The lesions were mostly located in cerebral lobes. Except 1 case which had operated, all of the cases were dignosed by steriotactic biopsy. All of the cases were diffuse large B-cell lymphoma, which were confirmed by immunohistochemistry. In one case clonality was showed by PCR. Two cases were not recieved any treatment because they rapidly proggressed to death after the diagnosis. Ten cases received combined treatment of chemotherapy and radiotherapy. The others were treated with radiotherapy or chemotherapy alone. Two cases were excluded from the survival analysis as the absence of the follow-up data. Six cases were died of the disease during the therapy in a period of ranging from 1 month to 24 months after the diagnosis. Five cases were still alive with a follow-up period ranging from 6 months to 48 months. Coclusion: PCNSL is very agressive tumor despite the combined therapy regiments. Steriotactic biopsy is very valuable diagnostic procedure as it is less invasive procedure, especially for deeply located lesions and it provides rapid diagnosis for this type of tumor which is treated non-surgical therapies. Besides molecular methods may be used for conforming the clonality of the tumor, especially for differential diagnosis.

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Molecular Pathology PP4-226 AN ALL-IRELAND CANCER BIOBANK AND INFORMATICS NETWORK FOR COLLABORATIVE TRANSLATIONAL RESEARCH Eoin Gaffney1, Ciaran Flanagan2 1 St James's Hospital and Trinity College Dublin, Ireland 2 I. D. E. A., Dublin, Ireland

INTRODUCTION: Translational research in cancer requires a large tissue sample volume and linked patient data. In Ireland, as in other European countries, cancer samples for research have not been collected with due quality control or stored consistently in accordance with standardised procedures. Insufficient sample aliquots have mitigated against the possibility of collaborative research. We therefore decided to develop an all-Ireland cancer biobank network with common informatics, to provide an infrastructure for collaborative translational research in the main hospitals of Ireland, north and south. METHODS: The Spanish Tumor Bank Network www.cnio.es was considered the most appropriate model for the Irish network, which includes two jurisdictions. The network concept was promoted among numerous stakeholder groups, including professional bodies, hospital management, medical staff, researchers, patient advocate groups, industry, and the public. Biobank Ireland Trust www.biobankireland.com was formed for public awareness and education, for fundraising, and to enable development of a restricted access online sample and informatics resource for potential research projects. No grants were sought, and no full-time personnel were assigned to the project. Disseminations included formal proposals and (international) presentations (ISBER 2005), and hosting and attending biobanking conferences. RESULTS: Progress has been slow and fitful! Informal networks of international biobanking experts were invaluable. Presentations, fundraising events, discussions with members of the public, and media coverage were all helpful. Also of significance was the inclusion in Ireland’s national cancer strategy (2006) of an aspiration to develop a national cancer biobank infrastructure. Biobank Ireland Trust’s business proposal forms the basis of the Irish network’s proof of concept phase: the network will commence in two hospitals, following which it will be extended to 10 others. The estimated cost of the network, excluding capital costs, is €8-10m. over 6 years. COMMENT: A biobank network is more about people than science. Patient advocate groups and the public must be consulted at the outset. When biobanking becomes standard of care in pathology, a biobank network will improve cancer care directly and indirectly. The Irish network will be a template for research infrastructure in other human diseases, will facilitate collaborations with other countries and will enable provision of biobanking assistance to developing nations. PP4-227 CYCLOOXIGENASE IN THE NORMAL HUMAN TISSUES – IS COX-1 REALLY A CONSTITUTIVE ISOFORM, AND COX-2 AN INDUCIBLE ISOFORM? Katarina Odar1, Maja Jerše1, Tomaž Zupanc2, Nina Zidar1 1 Institute of Pathology, Medical Faculty, University of Ljubljana, Ljubljana, Slovenia 2 Institute of Forensic Medicine, Medical Faculty, University of Ljubljana, Ljubljana, Slovenia

BACKGROUND: Cyclooxygenase is a key enzyme in prostanoid synthesis. It exists in at least two isoforms: cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). Despite the fact that COX isoforms have been widely studied in various pathological conditions, the data on their distribution and role in the normal tissues is still controversial. It is generally accepted that COX-1 is constitutively expressed in normal tissues as a housekeeper enzyme maintaining tissue homeostasis, whereas COX-2 is normally not present in most tissues, but can be induced by

various stimuli. Some studies, however, suggest that distribution and function of COX isoforms is more complex. We therefore analysed the distribution of COX isoforms in the normal human tissues. METHOD: Our study included autopsy samples of various organs and tissues from 10 healthy trauma victims. Immuno-histochemistry was performed by a peroxidase-streptavidin method on formalin fixed, paraffin-embedded tissue, using monoclonal antibodies against COX-1 and COX-2. In addition, western blot analysis of COX-2 protein expression was performed on frozen tissue samples of the brain, heart, kidney, liver, lung, thyroid, adrenal gland, testis, aorta and coronary artery. RESULTS: Using immunohistochemistry, we found COX-1 expression in blood vessels (in endothelial and smooth muscle cells) in all tested tissues, and in occasional inflammatory cells and macrophages, particularly in mucosal membranes. COX-2 was widely expressed in the parenchymal cells of endocrine glands (adrenal cortex, pituitary gland, thyroid, pancreas), testis, liver, central nervous system, kidney and in the glands of the gastric and colonic mucosa. COX-2 was focally expressed in the lung, and almost absent in the spleen and prostate. In the heart, occasional positive cardiomyocytes occurred with increasing age. In all organs and tissues, we observed COX-2 positivity in ganglion cells, occasional inflammatory cells and macrophages and focally in endothelial cells of blood vessels. With western blot analysis, we found detectable levels of COX-2 protein in all the tissues tested, presenting with bands of different intensities. CONCLUSION: Our results indicate that COX-2 is widely expressed in the parenchyma of many human organs and tissues, whereas COX-1 is predominantly found in blood vessels. Therefore, tissue distribution and role of COX isoforms under physiological conditions is clearly more complex than generally believed. We will probably have to revise the concept of “constitutive” and “inducible” COX isoforms. PP4-228 ERK AND PERK EXPRESSION, B-RAF MUTATIONS AND MICROSATELLITE INSTABILITY IN COLORECTAL CANCER Angelika. A. Saetta, Fanie Gigelou, Penelope Korkolopoulou, Nikolaos Kavantzas, Georgia Levidou, Polyanthi Papanastasiou, Angeliki Stamatelli, Maria Karlou, Nikolaos V. Michalopoulos, Efstratios Patsouris 1st Department of Pathology, School of Medicine, National and Kapodistrian University of Athens, Greece

Background: The RAS/RAF/MEK/ERK signaling pathway plays a pivotal role in controlling cellular growth, differentiation and survival mediating cellular responses to growth signals. Activating B-Raf mutations which stimulate RAF/MEK/ERK kinase cascade resulting in phosphorylation and subsequent activation of ERK, have been associated with colon cancer exhibiting microsatellite instability (MSI) -an alternative pathway of carcinogenesis caused by defective mismatch repair (MMR) system. The aim of this study was to determine the B-Raf mutational status and to asses the immunohistochemical pattern of ERK and activated ERK (pERK) in relation to MSI and hMLH1 promoter hypermethylation, of patients with colon cancer so as to elicit any potential associations with prognostic value. Methods: B-Raf exon 11 and 15 mutations were investigated by single stranded conformation polymorphism (SSCP) and DNA sequencing in 94 patients diagnosed with colon cancer. Additionally, immunohistochemical expression of both total ERK and pERK was examined. The presence of MSI was determined by analysis of sensitive mononucleotide markers (BAT-25, BAT-26) and the methylation status of hMLHI promoter was assessed by methylation specific PCR. Results: Total ERK cytoplasmic or nuclear immuoreactivity was detected in 92.7% (51/55) and 85.4% (47/55) of the cases respectively. Nuclear pERK immunoreactivity was found in 71.7% (32/45) while 11 (24.4%) cases displayed cytoplasmic positivity. There

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was no correlation between nuclear or cytoplasmic ERK LI and the presence of nuclear and/or cytoplasmic pERK immunoexpression. pERK nuclear immunoreactivity was positively correlated with tumor grade (p=0.0133) and Duke’s stage (p=0.09), whereas the latter was of marginal significance. B-Raf mutations in exon 15 were found in 12 cases. The presence or the score of ERK and pERK immunoexpression, either nuclear or cytoplasmic was not correlated with B-Raf mutations, MSI status or hMLH1 promoter methylation. Conclusion: Our findings indicate that ERK and activated ERK (pERK) are frequently expressed in colorectal cancer, independently of B-Raf status. Moreover nuclear pERK expression correlated with tumor grade and Duke’s stage, suggesting a possible further implication in tumor aggressiveness. The project is co-funded by the European Social Fund and National Resources-(EPEAEK II)-PYTHAGORAS II. PP4-229 DOUBLE IMMUNOSTAINING METHOD. WHAT IS THE SIGNIFICANCE? Naziye Ozkan1, Fulya Cakalagaoglu1, Emine Salva2 1 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department, Istanbul, Turkey 2 Marmara University, Medical Faculty, Pathology Department, Istanbul, Turkey Background.Immunostaining is a technique for identifying cellular or tissue antigen. Immunohistochemistry has been part of the routine technique of the pathology laboratory for over 20 years.These technique have significantly expanded the capabilities of the pathologist in diagnostic procedures.Immunohistochemical techniques were very sensitive and specific methods which utilize antigen-antibody complexes.Among the immunostaining methods the enzyme labelled streptavidin-biotin method is now widely used in routine testing. In study, immunohistochemical localization of -Smooth Muscle Actin ( -SMA),Proliferative Cell Nuclear Antigen (PCNA) was performed using the streptavidin-biotin peroxidase (Str.ABC/HRP) and streptavidin-biotin alkaline phosphatase (StrABC/AP) immunostaining methods.The double immunostaining method described may have wide application since it can be used both for paraffin sections and cytology. It does not require specialized technique or apparatus. The aim of our study to compare the immunoexpression of PCNA and -SMA in anti-Thy1.1 diffuse experimental glomerulonephritis rat model by Str.ABC/HRP,StrABC/AP and double (Str.ABC/HRP and Str.ABC/AP) immunohistochemistry methods (DIHC). Methods.Ten male Wistar Albino rats were divided into two groups of fifth: Group I Control(C): 5 rats treated with 0.1ml /100g normal saline intravenously (i.v) for four weeks; groupII glomerulonephritis (GN): 5 treated with anti-Thy1.1(0.25 g/100g) i.v at zero day. Kidneys from rats were collected as Instituional Animal Care and use Committee approved procedures.Representative tissue samples were stained with hematoxylin eosin (HE) and Gomori One Step Trichrome.In each case PCNA and -SMA expression were semiquantatitively scored on glomeruli and tubulointerstitium by Str.ABC/HRP,Str.ABC/AP and double immunohistochemical methods staining slides. Results:At the end of the study period morphological changes including tubulointerstitial injury and glomerular cell proliferations were significantly increased in glomeulonephritis group compared to control group.With in immunohistochemical methods,glomerular immunoexpression of

-SMA was similar in two groups.Tubulointerstitial immunoexpression of -SMA was similar in Str.ABC/AP and Str.ABC/HRP in GN compared to two groups,but different in DIHC.Immunoexpression of PCNA in all immunohistochemical methods on tubulointerstitial and glomeruli was significant increased in GN group compared to C. Conclusions.Double immunostaining was the method to detect two antigen in same site.That method was simple and reliable.

PP4-230 THE EFFECTS IN SKIN WOUND HEALING AND CELL PROLIFERATION OF THE PLASMID DNA ENCODING GM-CSF AND CHITOSAN Emine Salva1, Naziye Ozkan2, Fulya Cakalagaoglu2, Berna Karakoyun3, Julide Akbuga4 1 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department, Haydarpasa, stanbul, Turkey; Marmara University, Department of

Pharmaceutical Biotechnology, Faculty of Pharmacy, Haydarpasa, Istanbul, Turkey 2 Marmara University, Vocational School of Health Related Professions, Pathology Laboratory Department, Haydarpasa, Istanbul, Turkey 3 Marmara University, Nursing School, Haydarpasa, Istanbul, Turkey 4 Marmara University, Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Haydarpasa, Istanbul, Turkey Background: Granulocyte macrophage colony-stimulating factor (GM-CSF) regulates proliferation, differentiation and function of hematopoietic progenitor cells. Whereas a very important problem with the use of GM-CSF is that it has short biological t50 and it is necessary to administer frequent injections of high dose of GM-CSF, however this approach has been limited by the toxicity of this protein. Chitosan (CS), a natural cationic polysaccharide, has a high potential as a non-viral vector for gene delivery. Topical delivery of therapeutically relevant genes to wounded skin may help accelerate wound healing. The objective of this study was to investigate the encapsulation of hGM-CSF encoding gene into chitosan complex and in vitro transfection efficiency and therefore in vivo effects of GM-CSF complex application were identified in normally healing wounds. Method: pORF-hGM-CSF plasmid DNA was extracted by the alkaline lysis method.CS-pDNA complexes were prepared.In vitro transfection studies, the effect of the proliferative of the GM-CSF was determined by MTT test. We have used the assay to measure proliferative GM-CSF at the 24,48 and 72 hours. For skin topical application, Wistar rats were first anesthetized by ketamine, their backs shaved and 6 mm long punch biopsy was made at two individual sites on the dorsum of the back of each animal.Thereafter, 100 μg GM-CSF/CS complex (1:1,N/P) and chitosan per wound sites were topically applied.Skin samples were harvested 24,48,72 hours (acute term) and 1,2 weeks (chronic term) after application.Specimens were paraffin embedded, sectioned, stained using H&E and Gomori One Step Trichrome. Results: The better in vitro GM-CSF gene expression was found with 1:1 (N/P ratio). The dose of plasmid in complex is also important in protein expression. In vivo study, the neovascularization, epithelization, cellular content and vascularity increased in GM-CSF/CS complex group compared to chitosan group in acute term. In chronic term, the chronic inflammation, neovascularization, epithelization, cellular content, granulation tissue formation and vascularity increased in GM-CSF/CS complex group compared to chitosan group. In acute and chronic terms, the collagen deposition increased in chitosan group compared to GM-CSF group. Conclusion: According to our results, chitosan/DNA complex is thought to have ability to be a good potential in the treatment with GM-CSF gene so that it is to be a suitable delivery system for gene of chitosan, be applied from skin with chitosan of the gene and able to potential therapeutic effect to wound.

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PP4-231 FASCIN AND MIB-1 EXPRESSION IN SEROUS OVARIAN TUMORS Evanthia Kostopoulou1, Alexandros Daponte2, Rodoula Papamichali1, Athanasia Galani3, Maria Netsika3, Ioanna Chiotoglou4, Konstantia Zachou3, Dimitris Theodosiou3, George Koukoulis1 1 Department of Pathology, Medical School, University of Thessaly, Greece 2 Department of Obstetrics and Gynecology, Medical School, University of Thessaly, Greece 3 Department of Pathology, University Hospital of Thessaly, Greece 4 Laboratory of Cytogenetics and Molecular Genetics, Medical School, University of Thessaly, Greece Background: Fascin-1 is an actin-bundling protein that contributes to the formation of actin-based structures, including cellular surface protrusions that mediate cell movement. Studies in carcinomas suggest the emergence of fascin as a new prognostic indicator. Rare reports concerning fascin expression in ovarian carcinoma have not confirmed a clear-cut relation between immunostaining and established markers considered to reflect biological aggression. The aim of our study was to examine a group of serous ovarian tumors for possible relationships of fascin expression. The proliferative activity was also examined, since some studies suggested an inverse relationship between Ki-67 and fascin. Method: Fascin immunostaining was assessed in 70 specimens of serous ovarian tumors, with complete surgical staging when indicated, (45 carcinomas, 9 borderline tumors and 16 cystadenomas), retrieved from the files of the Pathology Department, University Hospital of Thessaly. Immunostaining was performed using two antibodies, IM20 (Novocastra, U.K.) and 55k-2 (Cellmarque, U.S.A.). Extent and intensity of immunoreactivity were semiquantitavely evaluated. Double staining for fascin and Ki67 was performed in carcinomas using the double Envision kit (Dako, Denmark) and MIB-1 antibody (DakoCytomation, Denmark). Western Blotting was performed on proteins isolated from tumor tissue kept at -80oC. Results: Fascin score was highest in carcinomas. Cystadenomas showed a score lower from carcinomas and borderline tumors (p<0.0001). Increased values (mean 4.70 vs 2) were observed in carcinomas of advanced stage (III/IV vs I) (p=0.04).Immunoreactivity was increased in metastases in comparison to primary tumors. Positivity was also found in the stroma of carcinomas, often intensified in the immediate vicinity. An interesting association between fascin and MIB-1 immunoreactivity was observed, the latter being lower in tumors showing increased fascin score. Conclusion: Our study showed that increased fascin immunoreactivity in serous ovarian carcinomas is associated with certain features of increased tumor aggressiveness. Increased score in specimens from metastatic sites suggests that fascin may be an important factor in ovarian carcinoma metastasis, as expected from its role in cell motility. We have also showed that cycling cells (MIB-1+) show lower fascin scores. Therefore, the actually invading cells, i.e. the “invasion fraction”, may differ from the “growth fraction” of a given tumor. Future studies could determine if fascin may become a helpful marker in gynecological pathology. PP4-232 SYNDECAN-1 AND VLA-4 INTEGRIN MOLECULES MAY CO-OPERATE IN MYELOMA CELLS UPON STIMULATION Vadim Baykov1, Tobias Slordahl2, Randi Utne Holt2, Anders Waage2, Anders Sundan2, Magne Borset2 1 Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway; Department of Pathology, St Petersburg State Medical University, St Petersburg, Russia 2 Department of Cancer Research and Molecular Medicine,

Norwegian University of Science and Technology, Trondheim, Norway; Department of Pathology, St Petersburg State Medical university, St Petersburg, Russia Background CD138 (syndecan-1) expression is a distinctive feature of plasma/myeloma cells in the bone marrow (BM). The integrin VLA-4 ( 4 1, CD49d/CD29)is the most common integrin on multiple myeloma (MM) cells. Both molecules are involved in adhesion of MM cells to matrix proteins and possibly in selective localization of multiple myeloma to the BM. Adhesion induces pro-survival signaling cascades, increases proliferation, promotes drug resistance and thus may contribute to tumor progression, although extramedullary spread of MM is believed to be associated with loss of cell adhesive properties. Adhesion of MM cells is substantially increased in vitro following stimulation by cytokines, mainly HGF,IGF-1 and SDF-1 . Existing data on CD138 and VLA-4 on MM cells with regard to clinical stage/progression are controversial, but soluble syndecan has been proved to be a strong negative prognostic factor. Possible interaction/cross-talk of integrins and syndecans is widely discussed; intriguing data have been demonstrated for carcinoma cells, but little evidence exists in MM so far. The aim was to study the expression and localization of syndecan-1 and VLA-4 on MM cells. Methods and Results 62 BM trephine samples from MM patients (9 pts in Durie stage I, 24 – st II and 29 – st III) were screened for CD138 and CD29 immunoreactivity. There was no significant difference between the groups. Precise localization of syndecan-1 and 4-integrins in the membrane of MM cells was studied by fluorescence laser scanning confocal microscopy (LSM-510, Zeiss, Germany) in INA-6 cells (MM cell line) following adhesion to fibronectin with or without stimulation with HGF, IGF-1 and SDF-1 . Scans were taken at the interface. In unstimulated cells 4 signal appeared as a central spot at the adhesion surface, surrounded by a ring-like CD138 signal, with clear separation of both signals in the majority of cells. Upon stimulation the signals from 4 and CD138 got co-localized in the majority of the cells in a ring-formed pattern, 4 moving laterally and merging with CD138-positive area. The same tendency was found in a primary MM cell sample. Conclusion This study is, to our knowledge, the first to indicate a possible co-operation between VLA-4 and syndecan-1 in MM cells. This involves lateral movement of VLA-4 molecules leading to co-localization of syndecan-1 and VLA-4 signals. Total syndecan-1 and VLA-4 integrin expression on MM cells has not been related to clinical stage in our setting. Further investigation is needed to elucidate possible role of the phenomenon in tumor progression. PP4-233 ACTIVATED ERK EXPRESSION IN RELATION WITH B-RAF GENE MUTATIONS IN UROTHELIAL BLADDER CARCINOMA Maria Karlou, Evmorfia Boltetsou, Angelica A. Saetta, Pollyanthi Papanastasiou, Georgia Levidou, Angeliki Stamatelli, Irene Thymara, Petros Pavlopoulos, Penelope Korkolopoulou, Efstratios Patsouris 1st Department of Pathology, Medical School, National and Kapodistrian University of Athens, Greece Background: Ras/RAF/MEK/ERK signaling pathway is commonly activated in human cancers. However the effect of ERK signaling on the prognosis of primary urinary bladder cancer (UC) is not clearly understood. Activating mutations in the serine/threonine kinase B-Raf are observed in several tumor types such as melanomas and thyroid, colorectal and ovarian carcinomas. The aim of this study was to examine the prognostic significance of phosphorylated ERK1/2 expression as a hallmark of ERK activation in relation with the presence of B-Raf somatic point mutations in Greek patients with primary UC. Methods: Tumor samples from 132 patients with primary urinary bladder cancer were examined for pERK1/2 immunohistochemical

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expression. PCR-SSCP and sequencing analysis were performed in 90 cases in order to detect activating mutations in exon 15 of the B-Raf gene. Statistical package STATA 9.0 for Windows was used for all statistical calculations. All results with a two-sided p

0.05 were considered statistically significant. Results: Nuclear immunoreactivity for pERK was detected in 99.2% (131/132) of cases, whereas cytoplasmic immunoreactivity in 97.7% of cases (129/132). The pERK nuclear expression was marginally related to tumor’s histological grade and stage (1/2 vs 3, Fisher’s exact test, p=0.082 and Ta vs T1/T2/T3 Fisher’s exact test, p=0.084). In univariate analysis for invasive carcinomas higher histological grade (p=0.0003), advanced tumor stage (p<0.0001) and high nuclear pERK expression (p=0.039) were associated with worse prognosis. However, when analysis was restricted in muscle-invasive carcinomas, advanced stage (p=0.001) emerged as the only factor adversely associated with survival. Multivariate analysis in invasive as well as in muscle-invasive carcinomas selected only tumor stage as significant prognostic factor (p=0.024 and p=0.039 respectively). Mutations in exon 15 of B-Raf gene were not detected in the examined cases. Conclusion: ERK activation as well as B-Raf mutations are frequently detected in certain tumor types implying their importance as potential targets for anticancer treatment. Our findings suggest that pERK expression is common and occurs in a B-Raf-independent manner in primary urinary bladder cancer. The correlation of pERK expression with high histological grade and advanced tumor stage suggests a potential implication in tumor aggressiveness while elevated nuclear pERK expression seems to be an adverse prognostic factor, although not independent of classical clinicopathological prognosticators. PP4-234 OVEREXPRESSION OF GLUTATHIONE SYNTHESIZING ENZYMES IN A MOUSE MODEL FOR DRUG-INDUCED STEATOHEPATITIS Pichler Martin1, Abuja Peter1, Aigelsreiter Ariane1, Mischinger Hans-Jörg2, Denk Helmut1, Zatloukal Kurt1 1 Institute of Pathology, Medical University Graz, Austria 2 Department of Surgery, Medical University Graz, Austria Background: Mallory bodies (MBs) are a characterictic, cytoplasmic inclusion found in hepatocytes in alcoholic steatohepatitis and other chronic non-alcoholic liver diseases. Oxidative stress participates in the pathogenesis of such disorders as an imbalance in cellular oxidant/antioxidant homoeostasis can influence inflammation, fibrosis and apoptosis. Glutathione is the major antioxidant in hepatocytes that serves several biological functions and alterations in the glutathione level induced by toxic factors may promote the development of chronic liver disease. Our aim was to study the glutathione metabolizing pathway in an animal model that shows features of drug-induced steatohepatitis. Methods: Livers of 3, 5-diethoxycarbonyl-1, 4-dihydrocollidine (DDC)-intoxicated swiss albino mice were harvested after 1 week DDC-treatment, 2.5 months DDC-treatment, 1 month DDC-free diet (=recovery) and 3 days DDC-refeeding. Total RNA was extracted and enzymes involved in glutathione metabolism were analyzed by Real-time quantitative RT-PCR. Results: Real-time PCR data revealed a deregulation of glutamate-cysteine ligase (GCL) and GSH synthetase (GSHS) in DDC-exposed murine livers. Compared to normal mouse liver, the enzymes GCL and GSHS were both overexpressed after 1 week (1.7 and 2.6 fold) and 2.5 months DDC intoxication (1.9 and 3.1 fold), respectively, but returned to a normal level after administration of a DDC-free diet for 1 month after 2.5 months of DDC intoxication (recovery). Refeeding recovered mice a DDC-containing diet for 3 days after one month of DDC-free diet induced GCL (1.6 fold) and GSHS (3.4 fold). Conclusion: Our data suggest an activation of the glutathione synthesis pathway in liver exposed to DDC intoxication. The increased expression of GCL and GSHS may be in part a compensatory effect for a depletion of homocysteine/cysteine in the affected liver, as these metabolites

are precursors in the glutathione synthesis and alterations of these metabolites are found in human ASH. Otherwise since oxidative stress is an important factor in the development of steatohepatitis, overexpression may be an indicator for increased oxidative stress defence in drug-affected hepatocytes. PP4-235 MOLECULAR AND IMMUNOHISTOCHEMICAL ANALYSIS OF P53 MUTATIONS IN BREAST CARCINOMAS Christianna Zachariou, Vassiliki Malamou-Mitsi, Angelos Skyrlas, Niki Agnantis Department of Pathology - Molecular Pathology Unit, Medical School, University of Ioannina, Greece Background.: Alterations of the p53 tumor suppressor gene are probably the most common genetic abnormalities in human malignancies. The majority of p53 mutations are missense and occur in the conserved DNA-binding domain (exons 5-8). In all, about 1150 different point mutations have been described, with more than 95% affecting exons 5-8. These mutations may lead to loss of DNA binding that is believed to be critical for the biological activity of p53. In breast cancer, frequency of p53 mutation is reported about 20%, lower than in other solid tumors. The implications for breast carcinogenesis however are unclear; in several studies it has been associated with more aggressive disease and worse overall survival. As direct sequencing of the gene is a time-consuming and laborious procedure, several prescreening methods have been developed. A modification of Denaturing Gradient Gel Electrophoresis (DGGE) known as GC-clamped DGGE appears to be more efficient and sensitive. Methods: DNA was extracted from 60 breast carcinomas of various histopathology and corresponding adjacent normal tissues. We used a PCR and GC-clamped DGGE assay, followed by sequencing, to investigate the presence of mutations within exons 5-8 of p53 gene. Additionally, we looked for p53 protein accumulation by immunostaining. Results: Out of 60 cases analyzed, 12 (20%) seemed to be carriers of a p53 aberration (7 in exon6, 2 in exon7, 2 in exon8 and 1 in exon5). The vast majority of mutations were documented in invasive ductal type-high grade tumors. In all (but one) cases that produced a variant DGGE-pattern, gene alterations were identified by sequencing and validated in TP53 Mutation Databases. Of these, two mutations: a missense at g.13418T>A (p.Y220N) and a deletion at g.13270delG have not been previously reported in breast cancer. The presence of mutations revealed strong correlation with high-grade breast carcinomas (p=0.002). Positive immunostaining of p53 protein was observed more frequently in invasive breast carcinomas (51%) than in in situ component (27%), with statistical significance (p=0.043). P53 overexression was associated with the presence of mutations in p53 gene (p=0.004). Conclusion: Our data indicate that PCR-DGGE is an efficient and highly sensitive prescreening approach for p53 mutation detection in DNA samples of breast cancer patients. The presence of mutations and p53 protein accumulation in invasive carcinomas, as well as the significant correlation between mutations and high tumor grade, reveal their possible implication in advanced stages of breast carcinogenesis. PP4-236 Withdrawn

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Soft Tissue, Joint and Bone PP4-237 HETEROTOPIC MESENTERIC OSSIFICATION: ULTRASTRUCTURAL FEATURES OF THREE CASES AND REVIEW OF THE LITERATURE Deborah Malvi1, Stefania Lega1, Christine Betts2, Maria Pia Foschini1 1 Section of Anatomic Pathology, University of Bologna, Bellaria Hospital, Italy 2 Department of Experimental Pathology, University of Bologna, Italy Background: Heterotopic ossification is a reactive metaplastic bone-producing process, which occurs in soft tissues mainly as a consequence of trauma. This reactive metaplastic process is well known in somatic tissues and named “myositis ossificans”. The histological finding of mature bone-tissue within the mesentery of the gastrointestinal tract is very uncommon and to our knowledge, less than 25 cases have been described in the literature. This kind of lesion may represent the soft-tissue counterpart of myositis ossificans and therefore, it has been designated under the term “heterotopic mesenteric ossification” (HMO). Purpose of the present paper is to describe the ultrastructural features of three cases. Method: Three patients were studied; they were male, aged 25, 48 and 58 years, respectively. All of them had a previous history of abdominal trauma and presented with intestinal obstruction. Results: Macroscopically, multiple, firm, greyish-white lesions were observed in the mesentery of small bowel in all cases and in one also in the omentum. Histologically the lesions were composed of steatonecrosis and haemorrhage surrounded by myo-fibroblastic proliferation, with spindle shaped cells, intermixed with an acute and chronic inflammatory infiltrate. At the periphery of these areas, eosinophilic material resembling osteoid was present. The “zone phenomenon” described in the myositis ossificans was observed. The myo-fibroblastic cells and osteoblasts were mitotically active, while they lacked atypia and atypical mitotic figures were absent. At ultrastructure, elongated cells immersed in the osteoid material were rich in rough endoplasmatic reticulum, contained vesicles and filaments and showed a well developed Golgi apparatus. The eccentric nuclei showed finely distributed chromatin with occasional prominent nucleoli. All these features are consistent with a secretory-like cell pattern and an active protein synthesis. Conclusion: The precise pathogenesis of HMO has still not yet been determined. Most probably it is related to metaplasia of mesenchymal stem cells, as a consequence of an exuberant reaction to trauma or injury, in predisposed individuals. The ultrastructural features observed here further support the hypothesis of stem cells actively producing osteoid material. PP4-238 EWING SARCOMAS AND PRIMITIVE NEUROECTODERMAL TUMORS SEEN THROUGH THE PRISM OF EQUITY Vesna Janevska1, Rubens Jovanovic1, Gjorgi Zafirovski2, Slavica Kostadinova-Kunovska1, Liljana Spasevska1, Milan Samardziski2, Gordana Petrusevska1 1 Institute of Pathology, Faculty of Medicine, Skopje, R. Macedonia 2 Clinic for Orthopaedic Surgery, University Clinical Center, Skopje, R. Macedonia Background: It is now well known that these two entities are in fact two faces of the same malignant tumor designated ES/PNET. Not so long ago there were slightly different treatment protocols for the two entities, giving rise to debates among clinicians and pathologists about “give me the precise diagnosis”. Methods: We analyzed 33 biopsy/operative archive specimens admitted at our institution from the Clinic for Orthopaedic Surgery, Skopje. All

tissue samples had been fixed in 10% neutral formaline and paraffin embedded (FFPE), prior to sectioning and staining with H.E. and monoclonal antibodies. Twenty three cases were diagnosed as ES and 10 as PNET, according to the microscopic morphology and immunohistochemical staining pattern for the following markers: Chromogranin, NSE, Synaptophysin, S-100 and CD99. We performed additional immunohistochemical stainings for p53, Bcl-2 and Ki-67. All of the immunohistochemical stainings were performed using the DAKO Cytomation EnVision+HRP system. We also isolated DNA from FFPE for subsequent identification of EWS/FLI1 fusion [t(11;22)(q24;12)]. For EWS/FLI1 gene fusion identification we used Hot start–Touch down-direct PCR on Auto Q server-thermal cycler. Primer sequences were constructed using Gene Runner v.3.05 on reference sequences obtained from Ensemble Database ver. 41.36c with start:pos.11/ex7EWS; end: pos.478/ ex6 FLI1. Results: 61% of the patiens with ES were males, versus 60% with PNET (p>0,05). The mean age of the patients with ES was 19,5 years (min. 7, max. 55, SD=12,33), versus 20,3 (min. 4, max. 60, SD= 17,51) in PNET (p>0,05). As for the immunohistochemistry, we encountered slightly more frequent (but not significantly) positivity of PNET cases for p53 (weak 1+ staining signal) – 30%, versus 13% of ES cases (p>0,05). In PNET there were only 1+ intensity signals detected for Bcl-2 (33% cases), versus 38% of ES. More intensive expression (2+, 3+) was found exclusively in ES cases (9,5% and 4,8% respectively), however, with statisticaly non significant difference (p>0,05). Ki-67 showed similar expression frequency in both ES (13%) and PNET (10%) with 1+ intensity (p>0,05). EWS/FLI1 gene fusion was detected in 43,4% ES, compared 30% of PNET cases (p>0,05). Conclusion: Except for the potentially differential staining patterns of ES and PNET for NSE, Synaptophysin and Chromogranin, we found no other significant differences between ES and PNET neither for immunohistochemical expression of p53, Bcl-2 and Ki-67, nor for the presence of the EWS/FLI1 gene fusion. PP4-239 A BRIEF REPORT OF SYNOVIAL SARCOMA CASES IN HACETTEPE UNIVERSITY: A MORPHOLOGICAL ANALYSIS Cigdem Himmetoglu, Havva Solak Ozseker, Gokhan Gedikoglu Hacettepe University Faculty of Medicine Department of Pathology, Turkey Background: Synovial sarcoma (SS) is a morphologically, clinically and genetically distinct entity. Unlike its name, it does not arise from or differentiate toward synovium and is issued under the name Tumours of Uncertain Differentiation in the recent WHO classification. SS is a mesenchymal spindle cell tumour with variable epithelial differentiation including glandular formation and has a specific chromosomal translocation t(X;18)(p11;q11). Method: This study aimed at re-evaluation of the SS cases in our institution’s pathology archive between 1990-2007 to create a brief idea about histopathologic features of SS and its relationship to prognosis. Results: From the 18 cases reported as malign mesenchymal tumor, some of which only favoring SS diagnosis, 15 were confirmed. More than half of the cases were male (8); the mean age at diagnosis was 32 years and there was a male predominance in the younger age group. Eight cases were consultation so macroscopic examination was available in 7 cases. Among those cases, the mean diameter of tumor varied between 2-10 cm. The most common locations were foot (3) and inguinal region (3), followed by upper extremity and head-neck region by 2 cases each, one case was located at the knee, the last 3 cases abdominal and lumbal. Excisional biopsy could be performed in 5 of the 6 cases operated in our institution and in none of the 5 cases in which lymph node dissection was done, metastasis was detected. Microscopically, 7 of 15 cases were biphasic, one consultation case was a monophasic ossifying SS located in heel. Tumor necrosis was present in 3 cases. An

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immunohistochemical panel composed of at least EMA and pancytokeratin was performed in all cases. At least focal positivity with one of these markers were used in addition to morphologic features. Mitotic count in 10 high power field ranged between 2-43; 3 cases with counts more than 10 / 10 HPF. Five patients received their adjuvant therapy in our institution, 2 patients received chemotherapy. Three patients received chemotherapy and radiotherapy, one due to lung metastasis 2 years after the first diagnosis and all died due to disease. Seven cases are alive without disease and the mean follow up period is 11 months. Conclusion: In the literature up to 50% of SS recur, usually within 2 years, nearly 40% metastasize, commonly to lungs and bone. Tumor free local excision followed by postoperative radiotherapy improves the survival. Young age, tumors <5 cm in diameter, with <10 mitoses / 10 HPF, SS18/SSX2 variant gene and no necrosis are good prognostic factors. PP4-240 OSTEOSARCOMA: REVIEW OF 107 CASES Nuray Kepil1, Pelin Bagci1, Sergulen Dervisoglu1, Didem Colpan2, Fazilet Dincbas2, Nil Molinas Mandel3, Murat Hiz4, Alp Ozkan5, Inci Yildiz5, Pembe Cagatay6 1 Istanbul University, Cerrahpasa Medical Faculty, Department of Pathology, Turkey 2 Istanbul University, Cerrahpasa Medical Faculty, Department of Radiation Oncology, Turkey 3 Istanbul University, Cerrahpasa Medical Faculty, Department of Medical Oncology, Turkey 4 Istanbul University, Cerrahpasa Medical Faculty, Department of Orthopaedic Surgery, Turkey 5 Istanbul University, Cerrahpasa Medical Faculty, Department of Pediatric Oncology, Turkey 6 Istanbul University, Cerrahpasa Medical Faculty, Department of Biostatistics, Turkey BACKGROUND: Osteosarcoma (OS) is the most common bone tumor of childhood and young adulthood. Despite new therapy modalities and therapy agents, prognosis is poor. Epidemiologic and genetic researches are the most promising studies to understand the nature of this tumor. METHOD: We reviewed 107 cases diagnosed in our department between 2000- 2007 according to the latest WHO classification; and analyzed them considering age, sex, etiology, localization, histological parameters (necrosis rates), recurrence, metastasis, and response to therapy. RESULTS: The patients were between 7-73 years; and the median age was 28. There was male predominance (59%). Knee localization was seen in 57% of the cases, and most of them (49%) were in the 3rd decade. After 3rd decade knee localization decreased gradually and from the 5th decade of life OSs were seen at the trunk and extremity parts except knee. Conventional and blastic OSs were the most common types (77%), and 23% of the cases were malignant fibrous histiocytoma like OSs. Metastasis and recurrence were mostly seen in males. Ninty-one percent of the patients had Huvos grade III-IV tumor necrosis. Metastasis occured in 15% of the patients and 10% of them recurred. Sixty-six percent of the recurred patients and 87% of the metastatic cases had Huvos grade I-II tumor necrosis. Epiphysis invasion was seen in 94% of the cases; but in 91% of them articular space was intact. There were 3 postradiotherapy OSs and an OS associated with retinoblastoma in our material. CONCLUSION: Age, sex and localization distribution of the cases are the same as the English literature. But the percentage of knee localization is fewer and upper extremity localization is slightly more in our review. Response to chemoradiotherapy is the most important factor that effects the metastasis and recurrence rates. Neoadjuvant chemoradiotherapy and surgery are the gold standarts of dealing with OS.

PP4-241 DIAGNOSTIC AND PROGNOSTIC SIGNIFICANCE OF AGNOR IN OSTEOSARCOMA, CHONDROSARCOMA, EWING SARCOMA AND GIANT CELL TUMOUR OF THE BONE Sibel Kayahan1, Fikri Oztop2, Gulcin Basdemir2, Dundar Sabah2, Nimet Karadayi1 1 The Ministry of Health Dr. Lutfi Kıidar Training and Research Hospital, Turkey 2 Ege University Medical Faculty, Turkey

Background: AgNOR method which is based on determination of nucleoli regulator regions (NORs) by silver staining is one of the cell proliferation markers and is useful for identification of prognosis as well as differentiation of benign and malign tumours. There are great advances on bone tumour differential diagnosis and establishment of prognostic parameters. Yet, the reason of different responses to chemotherapy at osteosarcoma (OS) and Ewing Sarcoma (ES) cases, sometimes non-existence of objective criteria to determine histological malignity degree at chondorsarcoma (CS), the reason of the aggressive behaviour of some bone giant cell tumours (BGCT) are certain questions waiting to be replied. Methods: In this study, 33 OS, 12 CS, 10 ES and 15 BGCT cases diagnosed at Ege University Medical Faculty Pathology Department are evaluated. Besides, 5 non-Hodgkin lymphoma cases are included for comparison with ES. Cytological preparations prepared by imprint method from tru-cut biopsies are fixed in ethyl alcohol. Silver staining is performed and slides are examined by light microscopy. AgNOR spots at 100 cell nuclei are counted and mean AgNOR value is calculated. Results: While OS cases have the highest AgNOR values, CS cases are on the second and ES are on the third rank. The lowest values belong to BGCT and AgNOR counts differ significantly among tumour groups. In addition, this method is helpful for differentiating ES and non-Hodgkin lymphoma or BGCT and OS. The histological grade of CS and AgNOR values have correlation since low grade tumours show less while high grade tumours show more AgNOR values. At OS cases where response to chemotherapy is higher than 90%, AgNOR values are also higher. Conclusion: AgNOR method has a limited value on differentiation of malign bone tumours as OS, CS, and ES. Although it is helpful for identification of response to chemotherapy at OS and prediction of metastasis at CS, such findings are not observed at ES. AgNOR seems to be useful for differentiating ES and lymphoma or BGCT and telangiectatic OS. At CS cases, it may be helpful for differentiation of low and high grade tumours. PP4-242 EXPRESION OF p53 AND Ki 67 IN ATYPICAL MALIGNANT CHONDROBLASTOMA Jelena Sopta1, Mirjana Atanackovic1, Vesna Mijucic2, Nenad Lujic3, Goran Tulic4, Dejan Minic1 1 Institute of Pathology, School of medicine,Belgrade,Serbia 2 Institute for Oncology, Belgrade, Serbia 3 Institute of Orthopedic Surgery “Banjica”, Belgrade, Serbia 4 Institute for Orthopedic Surgery, Clinical Center of Serbia, Belgrade, Serbia

Chondroblastoma is a rare primary, usually benign bone tumor, but aggressive behavior is described. During the 20 years we report from Register of bone tumors of the Institute of pathology Belgrade 5 cases with unexpected clinical and histological characteristics which pointed at atypical chondroblastoma. Four patients were female and one was male, the youngest was 17 , the oldest was 50. The middle ages were older compared to the patients with typical chondroblastoma. Clinical features were: pain, swelling and restriction of movement. The history of the disease varied from 3 months to 4 years. Location were :2 humerus , femur, tibia and coxa. Radiological studies showed lytic, destructive and expansive lesion. In 3 patients clinically was presented recurrent tumor, after surgical therapy. Biopsy

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tissue showed: fields with polyhedral tumor and giant cells, cartilaginous interstitial matrix, “chicken wire” calcification and cystic spaces with blood. But also increased cell and nuclear atypia, hyperchromasia, pleomorphism and abnormal mitotic figures pointed to aggressive chondroblastoma. Immunohistochemical studies revealed p 53 mutation ( in 3 from 5 cases) and extensive proliferate activity KI67 positive in more than 75% of cells (in 4 of 5 cases). Surgical therapy was applied. Two years later one patient with p53 positive, and KI67 positive tumor have radiological confirmed lesion to the lungs suspected to metastatic deposits. Other patients are with no evidence of recurrent or metastatic lesions. These findings suggest that expression of p53 and KI67 are not in the correlation with biological behavior of aggressive chondroblastoma. PP4-243 EXPRESION OF KI 67 IN MALIGNANT TRANSFORMATION OF CHRONIC PROLIFERATE FIBROBLASTIC PERIOSTAL LESIONS Jelena Sopta1, Mirjana Atanackovic1, Aleksandar Djordjevic2, Vesna Mijucic3, Vera Vojinovic3, Dejan Minic1 1 Institute of Pathology, School of Medicine, Belgrade, Serbia 2 Institute of Orthopedic Surgery “Banjica”, Belgrade, Serbia 3 Institute for Oncology, Belgrade, Serbia

On the Register of bone biopsy, Institute of Pathology, Belgrade was registered 2 cases of malignant transformation of chronic proliferate fibroblastic periostal lesions. In our opinion this lesions should be analyzed as separate entities in bone tumors pathology. Both patients were males. First patient, 24 years old BM had a slow growing tumor in proximal part of tibia with 5 years long anamnesis. After the first biopsy lesion was described as a proliferate, posttraumatic fibroblastic lesion of periost. Patient had 6 relapses durind the 13 years until the final diagnosis- osteosarcoma. Second patient, ST was 34 years old when was performed the first biopsy of femoral lesion after trauma. In the period of 9 years he had 3 relapses and repeted byopsis. The final diagnosis was again osteosarcoma ( low grade superficial). This cases have in common a few elements: sex of patients, long evolution of lesions (13, and 9 years), trauma in anamnesis, many relapses (6 and 3), primary diagnosis of proliferate periostal lesions and finally, lesions were firstly clinically manifested when patients were young adults (period of life when osteosarcoma is the most frequent). Radiography, CT scan, MR, scintigraphy and histopathology characteristics of lesions firstly pointed to fibroblastic periostal proliferate lesions with expressive osteoid production up to mature bone. After many relapses and repeated biopsies with identical morphological findings- no malignancy but intensive proliferation of periostal fiber tissue and production of mature bone arise transformation and lesions become low grade malignant tumors (low grade malignant osteosarcoma). Proliferate potential of this lesions was investigated by Ki67. Expression of Ki67 were les intensive in first biopsy and it become intensive in relapses, finale it was the most intensive in the last biopsy when was made diagnosis osteosarcoma. We conclude that increase of expression's intensity of Ki 67 prove to malignant transformations of posttraumatic proliferate fibroblastic periostal lesions. PP4-244 MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY OF 23 CASES OF DERMATOFIBROSARCOMA PROTUBERANS Radulescu Doinita1, Stolnicu Simona2, Jalba Lucia3, Dumitriu Sorin4 1 Department of Morphopathology, University of Medicine and Pharmacy ‘Gr. T. Popa ‘ Iassy, Romania 2 Department of Morphopathology, University of Medicine and Pharmacy Tg. Mures, Romania 3 Mavromati’ County Hospital, Botosani, Romania 4 St. Mary’ Children Hospital, Iassy, Romania

Aims: The dermatofibrosarcoma protuberans (DFSP) is a malignant soft tissue tumour, characterized by frequent recurrences and rarely metastases. The tumour occurs most commonly on the trunk, including chest, back, and abdominal wall. Less commonly, the neoplasm is located on the proximal extremities; it rarely involves the distal extremities. The head and neck, especially the scalp, are also commonly involved. The vulva and parotid gland are unusual sites of involvement. The tumour appears between 25 and 75 years old. DFSP presents under several morphological variants: fibrosarcomatous, myxoid, with giant cells, pigmented, myofibroblastic. This study aims, through the analysis of 23 cases of DFSP observed during 5 years, to specify the prognostic factors of these tumours. Methods: 23 cases of DFSP are viewed retrospectively. We made sections of paraffin blocks with hematoxylin-eosin and special histochemical stainings: P.A.S., Blue Alcian, Fontana, Trichrome of Masson. We also performed an immunohistochemical study using: CD 34, CD117, C Kit, S100 Protein, Factor VIII, CD 68. Results and conclusions: DFSP has a significant risk of local recurrence that depends on the local excision. A much higher recurrence rate is reported in tumours treated by superficial or incomplete excisions only. Local reccurence usually develops within three years after initial surgery. The global analysis reveals that the type of the variant, the cellular atypia, the mitotic index and necrosis are microscopic predictive factors for the evolution of the DFSP. PP4-245 RHABDOMYOSARCOMA: HISTOLOGICAL AND IMMUNOHISTOCHEMICAL ASPECTS Radulescu Doinita1, Stolnicu Simona2, Jalba Lucia3, Dumitriu Sorin4 1 Department of Morphopathology, University of Medicine and Pharmacy ‘Gr. T. Popa ‘ Iassy, Romania 2 Department of Morphopathology ,University of Medicine and Pharmacy Tg. Mures, Romania 3 Mavromati’ County Hospital, Botosani, Romania 4 St. Mary’ Children Hospital, Iassy, Romania Introduction: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of the childhood and adolescent periods and can seldomly be seen in adults too. Despite all research, a single prognostic classification for modelling the treatment is still needed. The clinicopathological features of our recorded rhabdomyosarcoma cases are reviewed and the major prognostic factors are discussed. Materials and method: We present 12 RMS cases located in chest wall, back, flank, abdominal wall, upper and lower extremities. Our patients were between 27 and 45 years old. We made sections on paraffin blocks with hematoxylin-eosin and special histochemical stainings: P.A.S., Van Gieson, Gomori. We also performed immunohistochemical reactions. Results and conclusions: Microscopically, we identified these RMS variants: embryonal, alveolar and pleomorphic. Desmin, Actin and Myogenin positivity of different intensities were observed in all cases. All the cases were treated by chemotherapy and radiotherapy whenever was needed. According to the clinical follow-up, the evolution was unfavorable when the tumour had more than 5 cm, when was located in upper and lower extremities, when was an alveolar or pleomorphic type and when had local recurrences. PP4-246 MULTIPLE GLOMUS TUMORS OF THE ANKLE WITH PROMINENT INTRANUCLEAR PSEUDOINCLUSIONS Jae Yeon Seok, Se Hoon Kim, Jieun Kwon, Yoon Hee Lee, Woo Ick Yang Department of Pathology, College of Medicine, Yonsei University, Seoul, Korea, Brain Korea 21 Project for Medical Science, Korea

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Glomus tumor is a neoplasm, composed of the modified smooth muscle cells of the glomus body. We report a case of multiple glomus tumors of the ankle which show varying histologic types: solid type (glomus tumor proper) and angiomatous type (glomangioma). Interestingly, the tumor cells of the present case show prominent intranuclear inclusions, a finding hitherto unreported in glomus tumors. Ultrastructural examination demonstrated that the nuclear inclusions are not true inclusion bodies but intranuclear cytoplasmic pseudoinclusions made by the cytoplasmic invaginations due to the deep and complex nuclear contours. PP4-247 COMBINED STUDIES CONCERNING IDENTIFICATION OF HIGH STRESS ZONES AT THE ACETABULAR BONE-BIPOLAR PROSTHESIS INTERFACE Dan Nelu Anusca1, Iancu Emil Plesea1, Nicolae Iliescu2, Vasile Nastasescu3, Mirela Corina Ghilusi4, Claudia Valentina Georgescu4, Oltin Tiberiu Pop1 1 University of Medicine and Pharmacy Craiova, Romania 2 University Politehnica Bucharest, Romania 3 Technical Military Academy Bucharest, Romania 4 Emergency County Hospital Craiova, Romania

Background The study is intending to determine the stress and strain state in the acetabular bone (AB) at the bone-bipolar prosthesis cup interface using combined numerical and experimental methods. Method AB stress state was analysed with the finite element method (FEM) and the photoelasticity on the plane models. For FEM, two calculus models were developed considering a frontal section (FP) and a sagital one (SP) through the upper part of a bipolar prosthesis and the surrounding zone of the AB. Numerical simulations were performed for both one and two legs standing positions (OLSP and TLSP) for both models. Main stress and equivalent stress fields distribution in the AB were obtained after processing of calculus results and the corresponding curves were plotted using a special post-processing program. With photoelasticity technique the isochromatic fringes field in the AB for the OLSP was recorded in a circular polariscope. Photoelastic data were processed and the distribution curve of the main stresses difference ( 1– 2) on the AB surface was plotted. Results In OLSP, in FP, pmax was 25MPa and pressure area (PA) was situated on the upper acetabular wall (UAW), near the acetabular rim (AR) with values decreasing towards the medial plane. In SP, maximal pressure (pmax) is much smaller (8MPa) and PA was situated towards the anterior aspect of the UAW with a more uniform distribution of the values. The resulting PA is ovoid, with the greater diameter in the sagital plane and shifted towards the anterior aspect of the UAW. The pressure values have a decreasing trend from lateral to medial. In TLSP, overall pressure values were much smaller that those of previous condition. In FP, pmax was 4MPa and the PA was restrained to a small area in the close proximity of AR. In SP, PA is larger than in FP, placed on both sides of median line and with values decreasing from posterior (3MPa) to anterior (1,5MPa). The resulting PA is like an antero-posterior narrow strip, near the AR with decreasing values from posterior to anterior. The results of photoelastic investigations indicated the same area with high level on the contact surface as numerical simulations. Conclusion The experimental study revealed that PA is placed on UAW near the AR and is varying both in shape and dimentions (decreasing in TLSP) and loading level (decreasing in TLSP compared with OLSP, from lateral to medial and from posterior to anterior).

PP4-248 MICROARRAY-COMPARATIVE GENOMIC HYBRIDIZATION OF ALVEOLAR SOFT PART SARCOMA. -GENE COPY NUMBER PROFILING AND IDENTIFICATION OF CANDIDATE GENES- Eun Ji Shon, Kyung Un Choi, Jee Yeon Kim, Mee Young Sol, Hyun Jeong Kang, Dong Hoon Shin, Ik Doo Kim, Won Young Park, Seong Muk Jeong, Jung Hee Lee Department of Pathology, School of Medicine, Pusan National University, Busan, Republic of Korea Background: Alveolar soft part sarcoma (ASPS) is a rare, histologically distinctive soft tissue sarcoma of unknown origin. Although ASPS is characterized by a specific alteration, der(17)t(X;17)(p11;q25), the entire spectrum of genetic events underlying the pathogenesis of ASPS is unclear. Method: Using microarray-based comparative genomic hybridization (aCGH), we examined DNA copy number changes of ASPS. aCGH composed of 4030 clones was performed with two cases of fresh frozen tumor tissue from 29 year-old male and 16 year-old female. Results: We identified 30 chromosome regions that were imbalanced in both tumors, and 11 of those regions were located on chromosome Xp (Xp11.3-p11.4, Xp21.1, Xp21.3, Xp22.11-p22.13, and Xp22.31-p22.33) Additional regions with increased copy number were observed at 1q25.1, 12p12.1 and 17p11.2. Loss was found on only one region, chromose 22q11.23. Several genes located within the amplified region of Xp included UTX, RPS6KA3, ZNF645, PHEX, USP11, PCTK1, UBE1, KAL1, ARSE, ARAF, SYN1, TIMP1, GYG2, BMX, ACE2, CXorf20, TMEM27, CA5BL, PDK3, PCYT1B, TMSB4X and XK genes. Conclusions: Our study showed genomic regions and new candidate genes that were associated with tumor pathogenesis of ASPS. PP4-249 AN UNINVESTIGATED CASE OF A MEDIASTINAL RHABDOMYOSARCOMA WITH MASSIVE PERICARDIAL EXTENSION Simion George1, Ilie Marius1, Barbulescu Catalin2, Marinescu Elena1, Sajin Maria1 1 Department of Pathology, Emergency University Hospital Bucharest, Romania 2 Department of Cardiothoracic Surgery, Emergency University Hospital Bucharest, Romania Background: Primary mediastinal rhabdomyosarcomas are extremely rare. The tumors can occur in adults and children, and follow a very aggressive clinical course. Method: A 27-year-old female, originated from an isolated ethnic region, was admitted in ER presenting cough, chest pain, and respiratory distress, superior and inferior vena cava syndromes that aggravated in the past 3 weeks. A CT scan of chest revealed a giant anterosuperior mediastinal mass with extension in the middle compartment, somewhat not well-defined borders, and inhomogeneous tissue structure. The tumor had a mass effect on the vessels at the basis of the heart with tracheal compression, left superior lobar bronchi obstruction, and pleural effusion in the left costodiafragmatic sinus. The clinical examination revealed a blood pressure of 80/ 40 mmHg, AV=126/min, turgescent jugular veins, bilateral lower limb edema, a dull sound at the percussion of the left hemithorax. Given the resectable appearance on CT surgical resection was undertaken, but the patient suddenly died of cardiac failure. Results: The autopsy of the corpse was made. The examination showed a giant anterosuperior mediastinal tumor with extension in the middle compartment, the compression of the large vessels at the heart basis, trachea and left lobar superior bronchi. The tumoral mass encircled the entire heart, which circumferential infiltrated the pericardium as numerous white nodules. Another findings: left superior lobe pneumonia, parapneumonial sero-fibrinous pleurisy, liver and renal stasis. Classical histological method and immunohistochemistry were used for pathological

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evaluation. Embryonal rhabdomyosarcoma composed in a great percentage of primitive cells devoid of rhabdomyoblastic differentiation, disposed in a very loose connecting stroma and rare cells with rhabdomyoblastic differentiation . IHC showed intensely positive reaction for vimentin, actin and weak positiving for desmin. Cytokeratin markers, CD20, CD34, CD68, CD117 and S100 are negative. Conclusion: Primary mediastinal rhabdomyosarcomas are extremely rare. In our 3 years old experience in collaborating with the Cardiothoracic Surgery Department this was the only soft tissue tumor from 16 mediastinal tumors. Since these tumors can remain clinically silent, they may have already reached large dimensions by the time of diagnosis. This is especially true for those masses originating from the chest wall. PP4-250 DESMOPLASTIC SMALL ROUND CELL TUMOR-PROBLEMS OF DIFFERENTIAL DIAGNOSIS IN UNUSUAL LOCATIONS Chmielik Ewa1, Nasierowska-Guttmejer Anna2, Lange Dariusz1, Nikiel Barbara1, Bialas Monika1 1 Department of Tumor Pathology, M. Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Gliwice, Poland 2 Department of Pathology, Clinical Hospital Ministry of Internal Affairs and Administration, Warsaw, Poland Background: Desmoplastic small round cell tumor (DSRCT) is well defined neoplasm with distinct clinical, pathologic, immunohistochemical and molecular features. This tumor shows epithelial, mesenchymal and neural differentiation. DSRCT occurs mainly in peritoneal cavity of young males but can occur in unusual locations and then diagnosis could be difficult. The aim of the study is analysis histopathological and immunohistochemical features of primary, recurrent and metastatic DSRCT in soft tissue, cardiac muscle and cervical lymphnode locations and to make a detailed differential diagnosis. Method: three cases of DSRCT in unusual locations were found in our consultant files. We studied routine H&E-stained sections and immunohistochemically-stained sections (AE1/AE3, EMA, CK19, CK20, vimentin, desmin, WT1, CD56, CD99, synaptophisin, NSE, CD56, actin, SMA, Myf 4, Myo-D1, S-100, Melan A, HMB-45, TTF-1 and LCA). Results: All of the studied cases pertained to young adult men of ages: 25 (case 1), 27 (case 2) and 23 (case 3). The 25 years-old patient presented with mass in the left popliteal fossa and recurrent tumor after 9 months. The 27 years-old man presented with a neoplastic infiltration in the cardiac muscle. The third 23 years-old patient showed metastatic bilateral cervical lymphnodes. Tumors were made up of small nests and well-defined islands of cells separated by thin strands ( case 1, case 3) or by broad desmoplastic strands ( case 2). The neoplastic cells were small- to medium-size with scanty or focally clear, vacuolated cytoplasm. There was evidence of rosette formation in the case 1 recurrent tumor as well as glandular arrangement in the case 2 tumor. Necrosis was noted. The tumor cells of case 1 and case 3 showed typical immunophenotype of DSRCT- AE1/AE3, desmin, NSE possitivity accompanied by EMA, vimentin, WT1, CD56. The cells displayed diffuse cytoplasmic and membranous reaction for CD99. The tumors cells of case 2 were positive for keratin, EMA, vimentin, WT1, CD56, NSE, but negative for desmin. Considering their unusual location, the tumors presented herein yielded broad differential diagnoses including Ewing’s sarcoma/PNET, alveolar and embryonal rhabdomyosarcoma, poorly differentiated synovial sarcoma, lymphoma, metastatic melanoma, metastatic small cell carcinoma, metastatic neuroendocrine carcinoma and Merkel cell carcinoma. Conclusions: DSRCT must be kept in mind when diagnosing primary tumors of soft tissue and metastatic lymphonodes in young men.

PP4-251 PRIMARY MYXOID TYPE LOW GRADE CHONDROSARCOMA OF THE XYPHOID PROCESS IN A YOUNG WOMAN Mattheos Bobos1, Evangelia Athanasiou2, Dimitrios Hatzibougias1, Athanasios Rigas3, Ioannis Hatzibougias2 1 Department of Pathology, Aristotle University Medical School, Thessaloniki, Greece 2 Department of Pathology, Faculty of Health and Care, Alexandrian Technological Institute, Thessaloniki, Greece 3 Surgery Private Practice, Greece Primary chondrosarcomas (CHS) of xyphoid process are rare and commonly occurs in elderly individuals. Herein we present a case of primary myxoid grade I CHS in a young adult. A 28 years-old woman presented clinically with a slow growing painless swelling on anterior chest wall. X-rays and CT examinations revealed an area of radiolucency with variable mineralization, indistinct margins, approximately 5 cm, centered in the xyphoid process. Macroscopic evaluation of surgically resected specimen showed a lobulated mass with a lucent glistering appearance with gritty areas. Histologically, the tumor formed nodules that invade the surrounding fibroadipose tissue. The extent of cellular atypia was mild and mitoses infrequently detected. Focally, increased cellularity was observed. The stroma was predominantly myxoid. A small areas of tumor necrosis was also found. It is important for the correct diagnosis of primary CHS to stress the attention for clinicopathological correlation of all the available data before arriving at a diagnosis. PP4-252 CASE REPORT : ATYPICAL GRANULAR CELL TUMOR Selvinaz Ozkara, Ilkay Tosun, Onder Peker, Pembegul Gunes, Gulistan Gumrukcu Department of Pathology, Haydarpasa Numune Education and Research Hospital, Turkey BACKGROUND: Granular cell tumor (GCT) is an uncommon soft-tissue tumor that is usually benign and readily treated with complete resection. It is found more commonly in the dermal-hypodermal tissue of the head, neck, chest wall and upper extremities. Apart from GCT s also have been observed in sites of previous trauma, such as surgical scars, and are associated with inflamation. We desciribe herein a GCT observed at the site of operation scar on the chest wall. CASE: The lesion was detected in a 69-year-old woman, located at the region of coronary artery bypass surgery scars on the chest wall. This operation was about 1 years ago. The lesion was painless subcutaneous nodule, which gradually enlarged. Excisional biopsy was performed. Histologically, there was a tumor, in the dermis and subcutis . It consisted of agregates of polygonal and spindle cells, sperated by thin bundles of mature collagen. The cytoplasm was eosinophilic and granular. Tumor cell nuclei was large and vesicular. There was no necrosis. mitotic activity was: 4-5/10hpf. Immunohistochemically, tumor was positive for S-100 protein and CD-68; negative for desmin, actin, HMB-45, CD-34, pancytokeratin. CONCLUSION : Malignant GCTs account for %1-2 of all GCTs. Because the malignant variant shares many striking pathologic similarities with reliable histologic criteria have been debated. Recently, Fanburg-Smith et al. classified atypical, malignant, and benign GCTs on the basis of six histologic criteria: necrosis, spindling, vesicular nuclei with large nucleoli, increase mitotic activity (>2 x10hpf), high nuclear to cytoplasmic ratio, and pleomorphism. Neoplasm that met three or more of these criteria were classified as atypical, and those that met one or two criteria were classified as atypical. The present tumor showed spindling and increasing mitotic activity. It can be classified as atypical GCT, suggesting malignant potential. Herein, we report this rare case of atypical GCT and present a brief review of the relevant literature.

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PP4-253 HISTOLOGICAL, GENETIC AND MOLECULAR DATA IN 96 CASES OF SOFT TISSUE TUMOURS, DIAGNOSED ON IPO – PORTO (PORTUGAL CANCER CENTRE – OPORTO) Luís Afonso, Manuel Teixeira, Carlos Lopes IPO - Portugal

INTRODUCTION: Diagnosis of soft tissue tumours is complex. Electron microscopy 20 years before and immunocytochemestry (ICC)nowadays, introduced important and decisive data to evaluate the histogenesis and differentiation of many sarcomas. In recent years, molecular pathology, essentially chromosomic alterations and genetic mutations studied by PCR techniques and/or FISH, are being added to the routine methods of diagnosis. The aim of this work is to evaluate the contribution of molecular pathology to the diagnosis of soft tissue sarcomas (visceral sarcomas were excluded) in Oporto Cancer Centre during the last 5 years (from 2001 until now). MATERIAL AND METHODS - A total of 96 soft tissue tumours diagnosed in the last 5 years was included in this study, with exclusion of visceral cases such as GIST. The material used was obtained from surgical specimens in most of cases (82 cases), and also from tru-cut biopsies (12 cases) and, in a small number of children tumours, also from fine needle aspiration material (2 cases). In these tumours, the following immunocytochemical antibodies were used in routine diagnosis: cytokeratins, EMA, vimentin, SMA, desmin, Myo D1, S-100 protein, CD31, CD34, CD99, CD10, HMB45. Genetic and molecular techniques included in most of cases: FISH and identification of fusion proteins using RT-PCR from paraffin sections. In a small number of cases a fresh tissue specimen was available, and conventional kariotype was performed. RESULTS In 17 cases diagnosed as synovial sarcoma by histology, the characteristic molecular alteration was present in 12. In 1 a diagnosis of Ewing sarcoma could be done and in 4 cases no other diagnosis was suggested.In Ewing sarcoma, concordance between histologic and molecular diagnosis was found in 12 from 14 cases. Regarding liposarcoma, 3 cases classified as myxoid by conventional histology were reclassified as well differentiated, and 1 as synovial sarcoma, after genetic study. On other hand, 2 cases classified as lipoma in conventional histology were reclassified as well differentiated liposarcoma after genetic study. COMMENTS: Molecular pathology was important to clarify the diagnosis of some cases of synovial sarcoma, Ewing sarcoma, liposarcoma and alveolar rhabdomyosarcoma, whose histology and ICC were not typical. In other cases, such as embryonal rhabdomyosarcoma, myxoid chondrosarcoma and malignant peripheral nerve sheath tumour molecular pathology was not so helpful but was informative to exclude other diagnosis and to give additional information whose real significance is not yet understood. PP4-254 CLINICAL AND PATHOLOGICAL ASPECTS IN PROXIMAL AND DISTAL FEMORAL PATHOLOGICAL FRACTURES DUE TO THE METASTASIS Sirbu Paul1, Radulescu Doinita1, Stolnicu Simona2, Petreus Tudor1 1 Emergency Hospital, University of Medicine and Pharmacy, Iasi, Romania 2 Department of Pathology, University of Medicine, Targu Mures, Romania

This paper has investigated a group of 55 patients with surgically removed metastasis in proximal and distal femur, by clinical, pathological and therapeutic means. The patient mean age was of 53,5 years. The fracture site was described on the femoral neck (5 cases), on the trochanteric region (14 cases), on the subtrochanteric region (20 cases) and on the distal femur (16 cases). Part of the cases (42 patients) showed history of different primary tumor localization (lung carcinoma – 19 cases, breast carcinoma – 12 cases, prostatic carcinomas – 8 cases, renal

carcinomas – 2 cases, larynx carcinoma – 1 case). All patients were operated by excising the tumor followed by cavity filling with acrylic cement and internal fixation, with immediate mobilization. In 52% cases the histopathological aspect of the metastasis was identical to the initial malignancy. In 13 cases where the primary malignancy was unknown, we have used histochemical methods to determine the tumor origin. Thus, we have determined more 4 primary tumors corresponding to the histopathological aspect of the metastasis (lung – 2 cases, breast – 1 case, ovary – 1 case). Despite the fact that rigid osteosynthesis provides the patient a psychological support by the early functional rehabilitation, the survival rate is influenced by the primary malignancy. PP4-255 CATEGORIZATION OF MUSCULAR DYSTROPHIES : AN IMMUNOHISTOCHEMICAL STUDY OF 44 CASES Elahe Keyhani1, Kimia Kahrizi1, Yousef Shafeghati1, Mojtaba Azimian2, Elham Darabi1, Hossein Najmabadi1 1 Genetics Research Center-University of Social Welfare & Rehabilitation Sciences, Iran 2 Neurology Department-University of Social Welfare & Rehabilitation Sciences, Iran

BACKGROUND-The term " Muscular Dystrophy" which was first used by Erb in 1891 referred to a group of muscle disorders manifest by progressive muscle weakness of the voluntary muscles as a result of primary muscle degeneration.The disorders are frequently hereditary. By now many subsets of the disorders were described, all with nearly same clinical and histopathological findings; so further examinations such as Immunohistochemistry on muscle specimens & using blotting methods are essencial for differentiating the disorders . In Iran the IHC method first established in 2004 and a Neuromuscular core fascility formed in Genetics Research Center since then for the referred patients. METHOD-Patients:Muscle biopsies were obtained from Deltoid muscle of the patients who were clinically diagnosed as muscular dystrophy .High serum CK & myopathic pattern in EMG have been reported for all.Normal muscle tissue was used as normal control.Antibodies:Three settings of Immunostainings were applied for each patient ;1-Dystrophin antibodies,2-Alpha,Beta&Gamma sarcoglycan and Dysferlin,3-Merosin. RESULTS-From all 44 patients;5 cases revealed complete absence of Dystrophins(DMD);10 with partially stained with Dystrophins(suggested for BMD);8 cases of Sarcoglycans deficiency;4 patients with Dysferlinopathy;2 cases with Merosin deficiency (CMD);14 cases revealed positive staining with all of the antibodies and should be followed by the others (Calpain-3,Caveolin...);finally one of the patients showed only severe necrosis & degeneration . CONCLUSION-According to our study IHC is an essencial technique for the diagnosis & differentiation of muscular dystrophies.It should be followed by the blotting methods in most of the cases(BMD & Dysferlinopathy);Molecular methods also should be performed for definite diagnosis in some other cases(subtyping of Sarcoglycanopathies). In our pilot study , the most frequent muscular dystrophies was Dystrophinopathies followed by Sacoglycanopathies .A study on a larger group of Iranian patients with more antibodies should be followed. PP4-256 INTRAABDOMINAL LOCALIZATION OF EXTRASKELETAL MYXOID CHONDROSARCOMA: CASE REPORT Adela Cimic, Semir Vranic, Nurija Bilalovic Department of Pathology, Clinical Center of the University of Sarajevo, Bosnia and Herzegovina

Extreskeletal myxoid chondrosarcomas with primary intraperitoneal localization are extremely rare with only several cases reported on Pubmed. We report a case of intraabdominal extraskeletal myxoid chondrosarcoma (EMC) in 57-year old

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woman that was surgically treated because of the expansive tumor mass in her abdomen. The tumor that measured 11x11x7 cm, was located below diaphragm. On gross examination, it was a multinodular, chondroid tumor, well demarcated with central necrotic and gelatinous area. Microscopically, tumor was composed of chondroblasts arranged in cords, within myxoid stroma. Some of the cells were binuclear and bizarre. Peripheral area was less cellular compared with the central area. Mitotic activity of tumor cells was low classifying this tumor as a low-grade soft tissue tumor. PP4-257 POLYVINYLPYRROLIDONE STORAGE DISEASE: A RARE CAUSE OF PATHOLOGIC FRACTURE Chien-Chen Tsai1, Wen-Chih Huang1, Chih-Hung Chang2 1 Department of Pathology, Far Eastern Memorial Hospital, Panchiao, Taipei, Taiwan 2 Department of Orthopedics, Far Eastern Memorial Hospital, Panchiao, Taipei, Taiwan

[INTRODUCTION] Polyvinylpyrrolidone (PVP) had been widely used as a plasma substitute or a retardent in some drugs decades ago. In Taiwan, PVP has been prescribed inappropriately as blood tonics for nutrition support for a long time. However, local injection or systemic administration of PVP-containing drugs could cause PVP to be retained in the reticulo-endothelial system and lead to PVP storage disease. [CASE REPORT] A 65-year-old Taiwanese lady was suffered from right hip pain after a fall in Nov. 2005. Normocytic anemia (Hb: 9-10 g/dl) was noticed. Image studies showed right femoral subtrochanter fracture. During operation, the fracture site exhibited some soft, yellowish, and gelatinous material within the marrow space; therefore, a bony tumor or metastatic lesion was suspected. Histologically, it showed sheets of blue-grayish vacuolated cells in a myxoid and hemorrhagic background with minimal inflammatory reaction. These vacuolated cells were positive for mucicarmine and Congo red stains, but negative for Peroidic acid-Schiff (PAS) and alcian blue stains. Immunohistochemically, these cells were negative for cytokeratin and S-100 protein, excluding the possibility of metastatic carcinoma or myxoid liposarcoma. Tracing back this patient’s medical history, she had been frequently given intravenous supplement of plasma substitutes as blood tonic at the local clinics for several years. The whole clinical and pathologic features suggested the diagnosis of PVP storage disease. [DISCUSSION] PVP storage disease, or PVP granuloma, occurred mainly in skin and soft tissue. Severe anemia and bony destruction were also noted in some cases, owing to massive infiltration of PVP storage cells in the marrow spaces. However, pathologic fracture due to PVP deposits in bones was rarely reported. For the pathologists, the differential diagnoses include metastatic epithelial tumors, such as signet-ring cell carcinoma and renal cell carcinoma. Chondromyxoid tumors of soft tissue, such as liposarcoma, chordoma, and chondrosarcoma, as well as other histiocytic storage disease, should also be considered. The characteristic histochemical staining results, combined with negative immunostains for cytokeratin and S-100, help reach the correct diagnosis. The clinicians and pathologists should be aware of this iatrogenic storage disease to prevent misdiagnosis. PP4-258 MONOSTOTIC PAGET DISEASE OF THE PATELLA Stephanos Milias1, Epaminondas Molyvas1, Abraham Ploumis2 1 Department of Pathology 424 General Military Hospital, Thessaloniki, Greece 2 Department of Orthopedics 424 General Military Hospital, Thessaloniki, Greece

Background: To present an unusual case of Paget Disease located at the patella. Method: The patient was female 46 years old, presenting with pain and swelling of the left patella. The clinical, radiographic and CT findings were more suggestive of Paget

disease and they did not show any other bone affected. The patient underwent a needle biopsy. Results: Microscopic examination revealed areas with prominent osteoclastic activity, while the osteoclasts were larger than usual, containing many nuclei per cell. There were also fewer osteoblastic areas with rimming of osteoblasts. No atypia was observed. The stroma was vascular and partially fibrotic. The bony trabeculae were disorganised, forming focally a “mosaic” pattern and few “cement” lines. The findings were suggestive of Paget disease and particularly of the osteolytic phase. Conclusion: Monostotic Paget disease of the patella is extremely rare. The etiologic factors of Paget disease are not clear yet. It seems that both genetic and non-genetic, perhaps viral, factors are assosiated with the disease. Molecular studies may contribute to the understanding of the pathophysiology of Paget disease and provide new therapeutical procedures in the future. PP4-259 OSTEOBLASTOMA-LIKE OSTEOSARCOMA. REPORT OF TWO CASES Didier Meseure1, Jean Marc Guinebretiere1, Sabah Boudjemaa2, Liliane Boccon Gibod2 1 Pathology Department, Centre René Huguenin, Saint Cloud, France 2 Pathology Department, Armand Trousseau Hospital,Paris,France

Background: Osteoblastic-like osteosarcoma (OLO) is a rare (1% of osteosarcoma) variety of osteosarcoma of low-grade malignancy with potential for recurrence when adequate surgical margins are not achieved and metastases. Differential diagnosis between osteoblastoma and OLO can be very difficult, because osteosarcoma may resemble osteoblastoma (clinically, radiologically and histologically), certain variants of osteoblastoma (agressive osteoblastoma, osteoblastoma with pseudosarcomatous features, multifocal osteoblastoma) may mimic osteosarcoma and osteoblastoma may rarely undergo malignant transformation. Distinguishing these two lesions is of great importance because their prognosis and treatment are different. In most cases, combination of clinical, radiological and pathologic features affords a clear distinction. The strongest histologic criteria to differentiate between them is tumoral permeation of the surrounding host tissue. Design and results:We report two cases of OLO in 8 year-old children (one male and one female). The first presented with a 7 months history of increasing pain in the right shoulder. A lacunar lesion of the first rib was resected, suggesting an osteoblastoma histologically. The lesion reccured in the subclavian region 18 months later and histological examination displayed a typical OLO. This patient underwent chemotherapy and radiotherapy. Three years later, there was a tumoral progression towards vertebrae and spinal tube, leading to the death of this patient one year later. The second patient had an osteolytic lesion located in the right iliac crest. The tumour revealed morphologic features of a benign-appearing osteoblastoma. Local reccurence occured one year later, owing to a wide curretage. Seven months later, a further extensive local reccurence had developped. An OLO was then diagnosed and the patient was treated with chemotherapy followed by en bloc resection of the iliac bone. She is now free of symptoms (8 months). Conclusions: Pathological diagnosis of OLO based on purely morphological parameters is difficult, especially on small biopsies. These two observations show that it is necessary to define clear morphologic and immunophenotypic characteristics for this low grade osteosarcoma often misdiagnosed as osteoblastoma. Alteration in suppressor gene p 53 localized on chromosome 17p13 seems to be an early event in the tumorogenesis of malignant osteoblastic tumours. Its characterisation might be helpful and need further investigations in these circumstances.

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PP4-260 MYOSITIS OSSIFICANS PROGRESSIVA: A CASE REPORT Ellouze Sameh, Mnif Lilia, Fakhfakh Ines, Ghariani Makki, Ayadi Lobna, Gouiaa Naourez, Mnif Jamel, Sellami-Boudawara Tahya Pathology Department. H. Bourguiba Hospital Sfax, Tunusia

Introduction: Myositis ossificans progressiva is a rare autosomal dominant progressive disease of connective tissue. It consists on a heterotopic osteogenesis frequently associated to characteristic congenital malformations. Case report: We report a case of a 24 year-old girl which presented a heterotopic bone formation leading gradually to ankylosis. The diagnosis was clinically suggested and confirmed when we’ve discovered bone bridges. Discussion: Congenital malformations, most commonly of big toes and thumbs, are important criterias to draw on the diagnosis earlier and to avoid unnecessary biopsy or resection which may trigger off a flare of the disease. This lesion simulates microscopically an extraosseous osteosarcoma and juxtacortical osteosarcoma with a highly cellular stroma associated with new bone formation. The most important diagnostic feature is the zonal phenomenon of maturation. Therapeutic measures are essentially preventive and the prognosis is poor. PP4-261 GIANT CELL GRANULOMA OF THE INFRA TEMPORAL FOSSA: A CASE REPORT Chtourou Imen, Bahri Ibtissem, Fakhfakh Ines, Kallel Rim, Ayadi Lobna, Hachicha Lilia, Gouiaa Naourez, Sellami-Boudawara Tahya Pathology Department; Habib Bourguiba Hospital, Sfax, Tunisia

Background: Giant cell granuloma (GCG) is an uncommon benign lesion that occurs almost exclusively whithin the jaw bone. Isolated cases of GCG occuring in the other cranial and facial bones have been documented. Location in temporal bone is extremely rare and less then 20 cases have been reported in the medical literature. Case report: A 46 year old man presented with a 3-year history of hearing loss on the left. Physical examination revealed a mass in the left preauricular area. Radiologic computed tomography showed a destructive multiloculated lesion of the left infratemporal fossa, the temporal and sphenoidal bones. This lesion eroded the left external auditory canal. Serum alcaline phosphatase, serum inorganic phosphate and calcium were all within the normal limits. A biopsy was performed. The microscopic examination of the specimen revealed a connective tissue stroma containing a large number of ovoid or spindle-shaped cells and multinucleated giant cells resembling osteoclasts. Foci of hemorrhage and deposit of hemosiderin were also present. There were some regular osteoid trabeculae. On immunohistochemical examination, both mononucleated and multinucleated cells expressed CD68 and focally S100 protein. Proliferate index assessed by KI-67 staining was estimated to 5-10%. The diagnosis of GCG was made. Conclusion: GCG is a rare benign lesion with distinct clinicopathological features. Modern neuro-imaging shows an expansile destructive multiloculated bone lesion mimicking a neoplastic mass. Histopathology is confirmatory of this benign lesion. Other giant cell containing lesions such as the giant cell tumour of the bone and a brown tumour of hyperparathyroidism need to be carefully excluded. Microsurgical excision is curative with a low recurrence rate. PP4-262 A MODIFIED STAGING FOR SOFT TISSUE SARCOMA –PROGNOSTIC INDEX (STS-PI) El Hindawi Ali1, Akl Maha2 1 Pathology Department Faculty of Medicine Cairo University, Egypt 2 Pathology Department Theodor Bilharz Research Institute,Egypt

This study was carried out on 122 patients suffering from different types of soft tissue sarcomas and ranging in their ages from 17 to 72 years. They were admitted to "Aswan Cancer Institute" in Egypt for surgical resection of their tumors. The resected specimens were proved to be soft tissue sarcomas of various microscopic types with a minimum follow up period of 60 months (5 years) . The aim of our study is to find out a reliable scoring system for grading and staging of such sarcomatous tumors that could reflect a convenient clinco-pathological prognostic index. Following proper gross morphological, histopathological and immunohistochemical diagnosis of the specimens under the study, they were given scores according to the following five parameters, as follows: (a) According to Tumor microscopic type: Score 1 indicates favorable prognosis (as in dermatofibrosarcoma protuberans) and Score 2: for other types of sarcomas.( b) According to Tumor longest dimension: Score 1: for tumors < 5 cm and Score 2: for tumors > 5 cm. (c) According to Tumor depth: Score 1: for Superficial tumors and Score 2: for Deep tumors. (d) As regards Tumor histological grade, according to the French Federation of Cancer Centers Sarcoma Groups (FFCCSG system) : tumors were given three scores; Score 1: for Grade I tumors (well differentiated), Score 2: for Grade II tumors(moderately differentiated) and Score 3: for Grade III tumors (well differentiated). Grading is best done in relation to histological morphological criteria including ;(degree of cellularity, pleomorphism, mitosis and necrosis), and finally the last parameter is (e) According to the presence or absence of Tumor metastases (LN or organ metastases); tumors were given either Score 0: for absent metastases and Score 6: when metastases are present. So our suggested STS-PI is as follows : The suggested soft tissue prognostic index (STS-PI) for the studied cases was calculated by summing up the previous scores relevant to each tumor and was found and described as 4 CLASSES as follows: -CLASS A : 4 (best score) -CLASS B : 5, 6, 7 (good score) -CLASS C : 8, 9 (medium score) -CLASS D : 10 – 15 (poor score) The 5 years survival rate for the studied tumors was recorded in 7/8 CLASS A tumors (87.5%), in 30/44 of CLASS B tumors (68.2 %) , and in 17/59 CLASS C tumors (28.8%) and was 0/11 in class D tumors (0 %), pointing to a reliable STS-PI. PP4-263 EWING SARCOMA, A RETROSPECTIVE STUDY Aleksandra Bonevski1, Dora Anzulovic2, Kristina Meljanac2, Luka Brcic2, Lovorka Batelja2, Iva Brcic2, Sven Seiwerth2 1 Children’s Hospital Zagreb, Zagreb, Croatia 2 Institute of Pathology, Medical School University of Zagreb, Zagreb, Croatia BACKGROUND. Recently, new concepts in Ewing sarcoma/PNET diagnosis and treatment were introduced. Since 1998, immunohistochemical verification of ES/PNET diagnosis, using CD99, is routinely performed at our Institute. Since end 2003, molecular methods (RT-PCR for EWS/FLI1) were introduced. In 2003, an algorithm was adopted including molecular analysis for residual disease and early recurrence. In order to establish a baseline patient population, we performed a partly retrospective and partly prospective analysis. PATIENTS AND METHODS. 78 Ewing sarcoma/PNET, diagnosed at the Institute of Pathology, Medical School University of Zagreb and treated at the Children’s Hospital Zagreb, in the period from 1990 to 2006, were included in this study. Of all analyzed tumors, 8 were extraosseal. Retrospectively, we submitted all collected tumor samples for both immunohistochemical and molecular analysis. Immunohistochemical analysis was successful in all analyzed samples. RT-PCR could not be successfully performed on samples older than 2000. From 2003, together 96 samples – tumor tissue, peripheral blood and 28 bone marrows were analyzed for characteristic translocations. RESULTS. Immunohistochemically, all tumor samples showed moderate to strong positivity, while EWS/FLI1 could not be demonstrated in

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6. From the samples submitted according to the new algorithm, 39 were positive by RT-PCR, among them 10 peripheral blood samples. In some cases multiple analyses of blood samples yielded only one positive. In two diagnostically problematic and immunohistochemicaly equivocal cases of small blue cell tumors molecular analysis fostered the final opinion. CONCLUSION. Introduction of molecular techniques enabled us to solve some diagnostic problems and to detect disseminated disease prior to clinical evidence producing a potent tool for improved patient care and disease management. PP4-264 DESMOID TUMORS OF THE CHEST WALL Olfa Ismail1, Leyla Abid1, Adel Marghli2, Jamal Ammar3, Tarek Kilani4, Faouzi El Mezni5 1 Department of Pathology, A Mami Hospital, Ariana, Tunisia 2 Department of Thoracic Surgery, A Mami Hospital, Ariana, Tunisia 3 Department of Pneumology, A Mami Hospital, Ariana, Tunisia 4 Department of Thoracic Surgery, A Mami Hospital,Ariana,Tunisia 5 Department of Pathology, A Mami Hospital, Ariana, Tunisia Introduction: Desmoid tumors (DT) are rare benign soft tissue neoplasms that arise from fascial or musculo-aponeurotic structures. Despite their benign microscopic appearance, they are characterized by invasion of contiguous structures and recurrence. Purpose: We retrospectively reviewed our institutional experience of DT of chest wall with emphasis on the clinical and pathologic characteristics. Material & Methods: Between January 1993 to January 2007, we retrospectively collected 6 cases of DT of chest wall. Medical records and microscopic slides were reviewed. Anti-bodies selected for immunohistochemical staining included vimentin, desmin, smooth muscle actin, S100 and CD 34. Results: There were 4 males and 2 females with mean age of 40 years. Only pediatric case was noted. Three patients had a history of previous trauma or surgery ( thoracotomy for lung adenocarcinoma and sternotomy for a aorto-coranory bypass). Complaints included pain (n=4), palpable mass (n=3), sensory symptoms and cough. In CT scan, lesions appeared as wall chest soft tissue tumor (n=4) with extension to the neck in one case. One tumor was located in posterior mediastinum mimicking neurogenic tumor and other was misdiagnosed as lung neoplasm. Surgical treatment was performed in 5 cases. The size of specimen varied from 3 to 14 cm and included ribs in 3 cases. Histology revealed a densely collagenated acellular lesion interspersed with areas of moderate cellularity consisting of fibroblastic and myofibroblastic cells. Mitotic figures were absent.There was no infiltration of surrounding structures. Chirurgical biopsy was performed in massive tumor with pleural, mediastinal and pulmonary involvement. Vimentin and smooth muscle actin were observed in all cases. S100 and desmine in one case. CD34 was not expressed by any of these tumors. One patient presented one recurrence and was treated by local resection and radiation therapy. Chemotherapy was beginned in a pediatric case. Discussion: Clinical presentation of chest wall DT in the literature is similar to our serie, but we not note a female predominance. The pathogenesis of DT is not clear. Trauma has been strongly implicated and recent work revealed hight incidence of B catenin gene mutations. Definitive diagnosis requires histopathology. Neurogenic tumors, fibrosarcomas, calcifying fibrous pseudotumors and localized fibrous tumors of the pleura should be considered in the differential diagnosis. CD34, smooth muscle actin and B catenin stains are useful in this setting.

PP4-265 CLINICO-PATHOLOGICAL AND OUTCOME FEATURES OF FIBROUS DYSPLASIA: RETROSPECTIVE STUDY OF A SERIES OF 26 CASES FROM TUNISIA Sonia Ziadi, Mounir Trimeche, Moncef Mokni, Sarra Mestiri, Badreddine Sriha, Sadok Korbi Departement of Pathology, CHU Farhat Hached, Sousse, Tunisia Background: Fibrous dysplasia is a rare condition in which defective maturation of osteoblasts results in abnormal bone formation. Is typically occurs in patients before 30 years of age with an equal ratio between males and females. FD presents in two forms: monostotic and polyostotic. The purpose of this retrospective study was to analyze clinical, histological and radiological characteristics and the outcome after treatment of this lesion in Tunisian population. Methods: Twenty six cases of histologicaly proven fibrous dysplasia diagnosed during a 15-year period from 1990 through 2005 at the Department of Pathology of the CHU Farhat Hached of Sousse, Tunisia. The following medical records were subjected to a detailed scrutiny: In patient clinical records, imaging studies, histopathological observations and outpatient clinical follow-up records. Results: In the present series male/female ratio was 1:4.2. Their ages ranged from 6 to 71 with an average age of 28.6. Monostotic fibrous dysplasia was found in 24 patients (92.3%), and 2 patients had polyosotic fibrous dysplasia (7.7%). Mc Cune-Albright syndrome and Mazabraud syndrome witch associating fibrous dysplasia were not noted in any case. The majority of the lesions were located in the upper maxilla and the mandible (65.4%). Imaging studies often show lesion with a ground glass matrix. Histopathological examination in the majority of these cases showed dysplastic bone surrounded by fibroblastic, some osteoblastic giant cells and hemosiderin filled macrophages. In the monostotic form, the clinical outcome was considered good in majority of cases; however, one patient has developed an osteosarcoma, three years later. In the two polyostotic cases outcome was less satisfactory because of fractures and deformities. Conclusion: This clinicopathological study revealed a female predominance so this can support the hormonal theory of fibrous dysplasia suggested by some authors. PP4-266 EGFR, P53 AND CAVEOLIN-1 EXPRESSION IN ADAMANTINOMA OF LONG BONES Ozgur Mete1, Bilge Bilgic1, Misten Demiryont1, Harzem Ozger2 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey 2 Istanbul University, Istanbul Faculty of Medicine, Department of Orthopaedic and Traumatology, Istanbul, Turkey BACKGROUND: Adamantinoma of long bones comprises about 0.4% of all primary tumors. In 90% of cases, tibia is the main site of involvement. World Health Organization defined it as a low grade, malignant biphasic tumor characterized by a different morphological patterns, most commonly epithelial cells, which are surrounded by a bland spindle-cell osteo-fibrous component. Nowadays, the epithelial part of the tumor is accepted to be neoplastic. Aneuploidy and p53 overexpression have only been demonstrated recently in one study. Since many studies have indicated an important role for growth factors and associated receptors in tumor growth we studied the expression of epidermal growth factor receptor (EGFR) and also p53 immunohistochemistry. Caveolin-1 is a cell membrane protein and regulate intracellular signal transduction. It is usually found in mesenchymal tissue and is believed to act as a tumor suppressor or tumor promoter in many tumors. However, its expression has not been investigated in adamantinoma of long bones. For this reason we added caveolin-1 immunohistochemistry to our study. METHODS: A total of 8 resected adamantinoma of the tibia were included in this study.

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All cases were classified according to the standard diagnostic criteria. Immunohistochemistry was performed by using anti-EGFR, p53 and anti-caveolin 1 antibodies. RESULTS: Nuclear p53, cytoplasmic and membranous Caveolin-1 and membranous EGFR reactivities were accepted as the positive immunostaining. Five cases (62.5%) showed focal and weak p53 expression only in epithelial areas. One case (12.5%) showed focal EGFR expression in the epithelial component. All cases (100%) showed strong caveolin-1 positivity in the epithelial part. Among them, 2 cases showed also focal caveolin-1 immunostaining in the fibrous component. Caveolin-1 expression was also found in endothelial cells. CONCLUSION: Although the percentage of p53 stained areas ranged from %10 to 30, our result supports that the epithelial part of the tumor is being neoplastic. As EGFR expression was present in one case it could not explain its relationship with tumor growth, but the down regulation of receptors may be one cause of this situation. Thus needs further investigations and correlations with epidermal growth factor expression. However, the high and strong caveolin-1 expression may be explained by its possible tumor promoter role in adamantinoma of long bones. Furthermore its predominant expression in the epithelial part of the tumor support the evidence that fibrous component of the tumor may be reactive. PP4-267 CAVEOLIN-1 AND C-KIT EXPRESSION IN 12 MYXOID/ROUND CELL LIPOSARCOMAS Ozgur Mete1, Bilge Bilgic1, Misten Demiryont1, Harzem Ozger2, Fulya Agaoglu3, Emin Darendeliler3 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey 2 Istanbul University, Istanbul Faculty of Medicine, Department of Ortopaedic and Traumatology, Istanbul, Turkey 3 Istanbul University, Institute of Oncology, Department of Radiation Oncology, Istanbul, Turkey BACKGROUND: Caveolin-1 is cell membrane protein and plays a major role in membrane traffic and signal transduction. Several studies have implied that caveolin-1 might have a tumor suppressor role; however, others have reported its tumor-promoting function. C-Kit expression has been utilized to identify patients with gastrointestinal stromal tumors (GISTs) who can be treated with imatinib. But its expression is not restricted to GISTs. Day by day, many tumors are adding to the c-kit positive tumors list and its expression in other soft tissue tumors is being an area of interest for therapeutic implications. Caveolin-1 and c-kit expression in sarcomas, especially in myxoid/round cell liposarcomas are not widely explored. For this reason, we decided to examine the expression of anti-caveolin-1 and c-kit antibodies in 12 cases of myxoid/round cell liposarcomas. METHODS: A total of 12 resected myxoid/round cell liposarcomas cases were included in this study. All cases were classified according to the standard diagnostic criteria. Three cases were composed of purely round cell liposarcoma and five cases were composed of purely myxoid liposarcoma. Four cases were composed of both myxoid and round cell liposarcoma. Immunohistochemistry was performed by using anti-caveolin-1 and anti-c-kit antibodies. RESULTS: The presence of membranous and/or cytoplasmic staining was accepted as a positive reaction for anti-caveolin-1 and c-Kit antibodies. Caveolin-1 expression was present in nine cases (75%) except three purely round cell liposarcomas and also in the round cell component of four cases containing both myxoid and round cell liposarcomas. Cytoplasmic c-Kit positivity was found in two cases. CONCLUSION: Round cell liposarcoma represents a histological continuum of myxoid liposarcoma. But the presence of round cell is a significance of unfavourable prognosis. Caveolin-1 expression has been investigated only in one recent study where focal and weak positivity was found in dedifferentiated liposarcomas. Our study results showed that caveolin-1 may have a potential role in the pathogenesis of

myxoid and round cell liposarcomas. Furthermore, its absence in the non-lipogenic primitive round cell component let us to think that its down-regulation may be a hallmark of poorer outcome. As c-Kit has been found in only 2 of our cases, further investigations with larger series are needed to illuminate its usefulness for the therapeutic implications. PP4-268 NEUROBLASTOMA IN ADULTS, PRESENTATION OF 2 CASES Hale Demir1, Ovgu Aydın1, Haydar Durak1, Sergulen Dervisoglu1, Mustafa Ozguroglu2 smet Sahinler3 1 Istanbul University, Cerrahpasa Medical Faculty, Department of Pathology, Turkey 2 Istanbul University, Cerrahpasa Medical Faculty, Department of Medical Oncology, Turkey 3 Istanbul University, Cerrahpasa Medical Faculty, Department of Medical Radiation Oncology, Turkey BACKGROUND: Neuroblastoma is one of the common malignant tumors of childhood but only about 100 cases have been reported in adults. The age range is 15-82 years with a median of 34 years. Male/ female distribution is equeal. Many of these tumors are reported in sites that are unusual for classical neuroblastoma (parotid, kidney, gluteal soft tissue, lower extremity). The survival rates are similar to those for INNS stage 3 and 4 childhood neuroblastomas. CASE 1: 50 year old male patient presented with paraoesophagial mass. Right thoracotomy was applied. 5x4x2 cm mass with a rib was excised and was sent to another pathology department, diagnosed as malignant tumor metastasis to paraoesophagial lymph node; and it was suggested that the tumor could be an osteosarcoma metastasis. The paraffin blocks were revised by Pathology Department of Cerrahpasa Medical Faculty. The diagnosis was neuroblastoma transformed from ganglioneuroma in paraoesophagial soft tissue confirmed by imunohistochemical staining with synaptophysin and chromogranin. CASE 2: 23 year old female patient presented with cervical mass. Two excision material, measuring 2x1.5x1 cm and 1.5x1x1 respectively were sent to another pathology center. According to the original pathology report, the materials were nodular and the cut surface was homogenous and yellow-white in colour. It was reported as neoplasm showing features of ganglioblastoma. The tumor cells were diffusely positive for NSE, focal positive with chromogranin negative for pancytokeratin and synaptophysin immunohistochemically. The paraffin blocks were revised by Pathology Department of Cerrahpasa Medical Faculty and diagnosed as neuroblastoma metastasis to lymph node. CONCLUSION: Neuroblastoma should be included in differential diagnosis of a small round cell tumor even in adult age especially in case of a background of schwannian stroma with scattered ganglion cells. This accompanying histology representing ganglioneuroma raises the possibility of dedifferentiation to neuroblastoma in the first case. The presence of an indolent form of a neuroblastoma is open to discussion for the second case. PP4-269 THYROID METASTASIS FROM PELVIC MALIGNANT SOLITARY FIBROUS TUMOR AFTER 13 YEARS OF RESECTION AND COEXISTENT PARATHYROID ADENOMA rem Paker1, Yuksel Kucuk Zeybek2, Gokhan Icoz3,

Bulent Karabulut2, Gulsen Kandiloglu1 1 Ege University, Faculty of Medicine, Department of Pathology, Turkey 2 Ege University, Faculty of Medicine, Department of Medical Oncology, Turkey 3 Ege University, Faculty of Medicine, Department of General Surgery, Turkey

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Solitary fibrous tumor is a rare spindle cell neoplasm which can be malignant. Here we report a 70 year-old woman presented with elevated calcium level during routine blood test. Her blood parathormone level was also increased. Ultrasonography of the thyroid showed the presence of multiple nodules and a parathyroid mass. Total bilateral thyroidectomy was performed. On gross examination of the thyroid, there were multiple nodules varying from 0.3 to 1.6 cm in diameter. Microscopically the largest nodule was well-circumscribed and composed of spindle cells. Immunohistochemically, the lesion was negative for CD34, CD68, CD117, EMA, pancytokeratin, TTF-1, smooth muscle actin, S-100 protein and positive for CD99. It was found out that the patient underwent laparotomy and a large pelvic mass was removed in 1993. This mass was diagnosed histologically as low grade malignant fibrous histiocytoma. She received radiotherapy and chemotherapy. In 2005, during the folow-up, lung and liver metastasis were detected by computed tomography scan. At the same time an abdominal mass was seen and biopsy of this mass revealed a morphology consistent with the previous tumor. Microscopically all specimens were reevaluated. Hemangiopericytomatous and storiform patterns were observed in the tumors. When we take the changes in the classification of soft tissue tumors into consideration, we diagnosed the tumor as malignant solitary fibrous tumor. As it is known, cellular form of solitary fibrous tumor can show malignant behaviour, in the form of recurrence and/or metastasis. The patient also received a diagnosis of parathyroid adenoma by the microscopic examination of the contralateral lobe and this mass was the main purpose of thyroidectomy. We reported this case because yet to our knowledge this is the first case of metastatic solitary fibrous tumor of thyroid coexistent with parathyroid adenoma. PP4-270 COX-1 AND COX-2 EXPRESSION IN OSTEOSARCOMA AND BENIGN BONE TUMORS Gulfiliz Gonlusen1, Aysen Erer2, Melek Ergin2, Seyda Erdogan2, Serdar Ozbarlas2 1 Cukurova University Medical Faculty Dept. of Pathology, Turkey 2 Cukurova University Medical Faculty Dept. of Pathology, Turkey Background: Cyclooxygenase (COX) enzymes catalyze the conversion of arachidonic acid to prostoglandins, which have a variety of physiologic and pathologic roles. Two COX isoenzymes have been described to date: COX-1, which is constitutively expressed by most tissues, and COX-2, which is inducible by cytokines, growth factors, and hormones. Overexpression of COX-2 has been discovered in a variety of solid tumors and pediatric sarcomas. In our study; we aimed to investigate the similarity and differences of COX-1 and COX-2 expression in benign bone tumors and osteosarcoma. Material and Methods: Paraffin-embedded tissue specimens from 35 bone tumors (20 osteosarcoma, 6 giant cell tumor, 6 fibrous dysplasia, 3 osteoblastoma cases) were retrieved from the files of the pathology department of Cukurova University, Medical Faculty. COX-1 and COX-2 were applied by immunohistochemical method. The results of staining for COX-1 and COX-2 were analyzed semiquantitatively by using an immunohistochemical scoring system (HSCORE) that combines the percentage of immunoreactive cells (quantity score) and an estimate of staining intensity (staining intensity score). Results: Strong (HSCORE:2-3)COX-1 expression was detected in five (33.4%) and negative (HSCORE:0) or weak positivity (HSCORE:1) in 10 benign cases (66.6%)and there was no strong positivity with COX-1 in osteosarcoma cases (P:0.005). Whereas; there was no significant differences with COX-2 between benign bone tumors and osteosarcoma cases (P:0.730). No correlation was found between COX-1 and COX-2 expression in osteosarcoma cases (r:0.15). Whereas, mild negative correlation was detected between benign bone tumors with COX-1 and COX-2(r:0.38, p:0.5). Conclusion:

We conclude that most cases of the benign bone tumors express COX-1 and COX-2 to varying degrees. Whereas there was no moderate or strong positivity with COX-1 in osteosarcoma cases. These results may be explained that cyclooxygenase enzymes do not play an important role in osteosarcoma pathogenesis. PP4-271 ACRAL MYXOINFLAMMATORY FIBROBLASTIC SARCOMA (REPORT OF 5 CASES) Esengul Uzuner1, Misten Demiryont1, Bilge Bilgic1, Harzem Ozger2 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Turkey 2 Istanbul University, Istanbul Faculty of Medicine, Department of Orthopedics and Traumatology, Turkey BACKROUND: Acral myxoinflammatory fibroblastic sarcoma (AMFS) was first described in 1998 as a rare tumor of distal extremities. AMFS is usually localized in the fingers within the subcutaneous tissues. It develops in patients of all ages with a median of late 40s. Grossly, it is an infiltrative multinodular mass measured 1-8 cm in diameter. AMFS consists of bizarre cells which have enlarged vesicular nuclei with inclusion-like nucleoli and large eosinophilic cytoplasms. The stromal component of the tumor contains various amounts of myxoid and hyalen matrix mixed with inflammatory cells. METHODS: We report 5 cases of AMFS, occuring in 3 males and 2 females with their clinical, histological and immunohistochemical features. The mean age was 45 (range 26-64). The tumor sizes were ranged from 1 cm to 6 cm (mean 2.8 cm). The localizations of the tumors were as follows: 2 ankles, 1 toe, 2 hands (D5 and extensor surface). Three patients presented with painless swelling, one with pain and mass and one with only pain. The symptom duration of cases was between 3,5 months and 4 years. Four cases, which were sent to us for consultation, were previously diagnosed as myxoid type malignant fibrous histiocytoma, chondrosarcoma, low grade fibrosarcoma and sarcoma (NOS). RESULTS: Immunohistochemical studies revealed positivity for vimentin (4/4), CD68 (5/5) and CD34 (2/3). Positive immune reaction was focally observed only in 2 of 5 cases. No positive immunostaining was detected for pancytokeratin (4/4) and smooth muscle actin (3/3). Ki-67 proliferation indices were between 2-5%. Four cases was treated with local excision, whereas 1 case with local amputation. Recurrence was seen only in one that was localized at extensor surface of the hand. None of the cases showed metastasis with a median follow-up of 17 months. CONCLUSIONS: AMFS is a very rare entity that must be kept in mind in the differential diagnoses of tumoral and non tumoral lesions, such as tenosynovitis, nodular fasciitis, giant cell tumor, inflammatory myofibroblastic tumor, liposarcoma, epithelioid sarcoma and malignant fibrous histiocytoma. Although we detected recurrence only in one case and no metastasis, AMFS is known to be a low-grade sarcoma that shows recurrence and has metastatic potential. PP4-272 HMGA1 REARRANGMENTS OCCUR IN A SMALL SUBSET OF SPINDLE CELL LIPOMAS Fabiola Medeiros, Xiaoke Wang, Michele Erickson-Johnson, Antonio Nascimento, Andre Oliveira Mayo Clinic, Rochester, MN, USA Background: Spindle cell and pleomorphic lipomas are benign subcutaneous adipose tissue neoplasms typically occurring in the neck and back of older men. Conventional cytogenetic analysis often shows monosomy or partial losses of chromosomes 13 and 16. Rearrangements of 6p21 have been reported in these tumors, and one study has identified HMGA1 overexpression by immunohistochemistry in a single example (Dumollard JM et al Ann Pathol 2001; 21:303-10). Because we have recently encountered a classic case of spindle cell lipoma in the face of a

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66-year-old man with rearrangements of chromosome 6p21, we hypothesized that the HMGA1, a gene occasionally rearranged in ordinary lipomas and other benign neoplasms – was involved. The aim of this study was to investigate the frequency of HMGA1 rearrangements in a series of 17 of such tumors. Method: Seventeen cases of spindle cell lipomas with typical histopathologic features were studied for rearrangements of HMGA1 and HMGA2 by fluorescence in situ hybridization (FISH) on paraffin-embedded sections. Fresh and frozen tissues were available from the index case for standard cytogenetic analysis and reverse-transcriptase polymerase chain reaction (RT-PCR). Results: Cytogenetic analysis was performed in one case and demonstrated the following karyotype: 46,XY, t(1;6) (p32;p21.3), del(13)(q12q14)[19]/46,XY[1]. FISH analysis showed a balanced rearrangement of HMGA1, and semi-quantitative RT-PCR confirmed transcriptional upregulation of this gene. Molecular cytogenetic studies of 16 additional spindle cell lipomas revealed a balanced rearrangement of HMGA1 in a single case. HMGA2 rearrangements were not found in any of the cases studied. Conclusion: Rearrangements of chromosome 6p21 with involvement of HMGA1 seem to occur in approximately 10% of spindle cell lipomas (2/17 in the present series), a frequency similar to that observed in ordinary lipomas. This finding suggests that HMGA1 may play a role in the pathogenesis of a subset of these tumors. PP4-273 HYDATID CYST OF BONE; REPORT OF FIVE CASES Aysen Erer1, Seyda Erdogan1, Melek Ergin1, Serdar Ozbarlas2, Gulfiliz Gonlusen1 1 Cukurova University, Medical Faculty, Pathology Department, Turkey 2 Cukurova University, Medical Faculty, Orthopedic Surgery Department, Turkey Background: Hydatid cyst is a parasitic infection caused by larval stage of Echinococus granulosus in man and domestic animals. The incidence of hydatid cyst in liver, lung, kidney and brain are 59-75%, 27%, 3% and 1-2% respectively. Hydatic cyst in bone is extremely rare and found in only 0.5-2% of cases. In endemic areas, this ratio can increase to 4%. Material and Methods: In a comprehensive study of 109 cyst hydatid cases from pathology files of Cukurova University Medical Faculty between 2002-2007 years; we identified five bone hydatid cysts (5.5%). Results: The age distribution was between 18 and 55 years. Three of them were male, two patients were female. Among the five cases; four cases were admitted to our hospital with pain and one patient with pathologic fracture. The localization of the lesions was femur (two cases), tibia (one case), iliac bone (one case), acetabulum (one case). Conclusion: In this study, we present five hydatid cyst in rare localization and review the literature. PP4-274 SKULL BASE CHORDOMAS A RETROSPECTIVE STUDY OF 14 CASES Faten Limaiem1, Ines Chelly1, Amina Mekni1, Salma Bellil1, Fathia Maamouri1, Slim Haouet1, Nidhameddine Kchir1, Mohamed Moncef Zitouna1, Mohamed Khaldi2, Khedija Bellil1 1 Department of Pathology. La Rabta Hospital, Tunisia 2 Department of Neurosurgery La Rabta Hospital, Tunisia Chordomas are uncommon malignant neoplasms arising from remnants of the notochord. They account for 1–4% of all primary malignant bone tumours and are mainly localized in the sacrococcygium (55%). Skull base chordomas are extremely rare representing less than 1% of all intracranial tumours. The aim of the present study is to analyze clinicopathological features of chordomas and to discuss differential diagnosis. Over the 12-year period from January 1997 to December 2006, 14 cases of skull base chordomas were diagnosed at the pathology department of La Rabta hospital. Relevant clinical information and hematoxylin

& eosin stained tissue sections were available for review in all cases. Clinical features, symptoms at diagnosis, treatment and outcome were retrospectively analyzed. There were 11 male and 3 female patients (sex ratio M/F = 3,67) aged between 19 and 72 years with a mean age at presentation of 46,57 years. The most frequent complaints were headache, diplopia and decreased vision. Prior to surgery, each patient was investigated with contrast-enhanced magnetic resonance imaging (MRI) and contrast-enhanced computed tomography (CT). Seven tumours involved the clivus, 3 were intrasellar, 2 were located in the cavernous sinus and 2 involved the sphenoid sinus. Surgical resection of the tumour was achieved in all patients. Pathological examination of the surgical specimen was compatible with classic chordoma in 12 cases and chondroid chordoma in 2 cases. Immunohistochemical study was performed in only 2 cases and showed positive immunostaining of tumour cells with cytokeratin, EMA, vimentin and S-100 protein. PP4-275 ANGIOMATOID FIBROUS HISTIOCYTOMA: REPORT OF THREE CASES WITH CYTOLOGICAL AND HISTOLOGICAL FEATURES Mine Gulluoglu, Semen Yesil, Dilek Yilmazbayhan, Misten Demiryont Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Turkey Angiomatoid fibrous histiocytoma (AFH) is an intermediate grade histiocytic tumor which occurs in children and young adults. In this case report, we present the cytological and histological features of three AFH cases. The patients were all male, and 28, 22 and 36 years old. The localizations of the tumors were supraclavicular, forearm and inguinal region, and their diameters were 10 cm, 4 cm and 2.5 cm, respectively. In the fine needle aspiration material of the supraclavicular mass showed highly cellular smears displaying polygonal and spindle shaped cells, either dispersed or forming three dimentional clusters in a lymphocyte-rich background. The tumor cells had ovoid or spindle-shaped nuclei, fine chromatin, small inconspicious nucleoli. The excisional biopsy material of all three patients showed similar histopathological features. The tumors were composed of multiple cellular nodules surrounded by fibrous pseudocapsule and contained blood filled cavities in a lymphocytic and inflammatory backgound. Groups of hemosiderin-laden macrophages were also present in varying amounts. All three tumors were positive for CD68 and epithelial membrane antigen, negative for endothelial markers and one of them was positive for desmin. The tumor localized in cervical area had been resected with indeterminate surgical margins and recurred in 6 months time. The reexcision material showed similar histomorphological features. AFH should be included in the differential diagnosis in a child or young adult patient having a slow-growing subcutaneous mass with cytological and histological features of an intermediate grade histiocytic mesenchymal tumor. The morphological features as well as immunohistochemical findings make the accurate diagnosis possible. PP4-276 CASE REPORT: MELANOTIC NEUROECTODERMAL TUMOR OF INFANCY Murat Sezak1, Basak Doganavsargil1, Yesim Ertan1, Saffet Mutluer2, Taner Akalin1 1 Ege University School of Medicine, Pathology Department Izmir, Turkey 2 Ege University School of Medicine, Neurosurgery Department Izmir, Turkey Background: Melanotic neuroectodermal tumor of infancy (MNTI) is a rare benign neoplasm that may be locally aggressive. Since the first description by Krompecher in 1918, approximately

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350 cases are reported in the medical literature. The tumor affects most commonly maxilla (%70) followed by the skull and mandible during first year of life. Early diagnosis and radical surgery are critical for long-term cure. Case presentation: We are presenting a 18 month old boy with a lump in the right temporal region since the age of 3 months. Because the tumor had a relatively fast grow, she was referred to our hospital. Radiological examination showed a tumor located in the right mastoid which was demarcated and radiolucent. It was not adhered to the dura mater. Tumor was completely excised by neurosurgeons. Microscopically, tumor have two components, one of them was composed of large polygonal epitelioid cells with deposits of melanin and the other component was smaller neuroblast-like cells characterized with round centrally located hyperchromatic nuclei and scant cytoplasm in a dense sclerotic fibrous stroma. Tumor cells were occasionally arranged as alveolar nests, small nests and solid sheets. Necrosis, hemorrhage, nuclear pleomorphism and mitoses were not evident. Immunohistochemically tumor was positive for HMB-45 and negative for synaptophysin, pancytokeratin, epithelial membrane antigen, protein S-100, glial fibrillary acidic protein, chromogranin A, ubiquitin. The postoperative course was uneventful. There was neither residual tumor nor recurrences at the control of 3., 6. and 15. months of post operation duration with radiologic evaluation (magnetic resonance images) . PP4-277 PRIMARY SCHWANNOMA OF LYMPH NODE IN THE RETROPERITONEAL REGION Deniz Peker1, I. Willis2, E. Saiz1

1 Department of Pathology and Laboratory Medicine, Mount Sinai Medical Center, Miami Beach, USA 2 Department of Surgery, Mount Sinai Medical Center, Miami Beach, USA Intranodal schwannoma is an uncommon benign tumor of lymph node. We report a case of an intranodal schwannoma in the retroperitoneum of a 61-year-old woman. The patient underwent left total nephrectomy for the retroperitoneal mass thought to involve the kidney. Macroscopic examination of the specimen revealed a well-circumscribed 4.5x4.0x4.0 cm nodule located in the perihilar fat. Microscopically, the nodule was composed of a proliferation of bland spindled cells, which were immunohistochemically strongly positive for S100 protein, and negative for Actin 1A4, HHF35, Desmin, Melan A, Cytokeratin AE1/3, CD 68, CD 34, and CD 45. The pathological findings led to a diagnosis of a very rare case of primary intranodal schwannoma presenting as a retroperitoneal mass.

Infectious Diseases PP4-278 DIGESTIVE ACTINOMYCOSIS, REPORT OF 4 CASES. Mestiri Sarra1, Trabelsi Amel2, Yacoubi Med. Tahar2, Ben Yacoub-Abid Lilia2, Hmissa Sihem2, Korbi Sadok2 1 Pathology Department, F. Hached Hospital, Sousse, Tunisia 2 Pathology Department, F. Hached Hospital, 4000 Tunisia Background: Actinomycosis is a chronic granulomatous infection, that is relatively rare, caused by a Gram positive anaerobic bacteria. Digestive locations usually simulate a malignant process. Diagnoses difficulties usually lead to a surgical resection. Method: We report four cases of digestive actinomycosis, located to the appendix, the coecum and to the pancreas in respectively, 2 cases, 1 case and 1 case. The appendic ular location presented clinically as an appendicular syndrome. The two other localisations simulated a neoplastic process. Results: The 4 patients underwent a surgical resection. Histopathological examination has leaded to the correct diagnosis, showing a granulomatous infiltrate, surrounding colonies of actinomyces. Penicillin therapy was indicated in the 4 cases with a favourable clinical outcome. Conclusion: Abdominal location of actinomycosis is a rare with poorly specific clinical and radiologic features, the diagnosis is often made on surgical resection specimens. Treatment is based essentially on a well adapted antibiotherapy. PP4-279 OCCURRENCE OF GASTRO-DUODENAL GIARDIASIS AND/OR HELICOBACTER PYLORI INFECTION AFTER GASTRIC ACID SUPPRESSION Omran Zeinab1, Moussa Mona1, Nosseir Mona1, Guirguis Nevine2, Yehia Hisham3, Elkhayat Hisham3, Siam Moataz3, Yousef Magdy3 1 Theodor Bilharz Research Institute Pathology Departments, Egypt 2 Theodor Bilharz Research Institute Parasitology, Egypt 3 Theodor Bilharz Research Institute Hepatology and Gastroenterology Departments, Egypt ABSTRACT Giardia lamblia (G. lamblia) is a common world-wide parasitic infestation. This study aims to detect gastric and/or duodenal G. lamblia colonization in patients receiving acid suppressing drug (Omeprazole) to assess its pathological repercussions on gastrointestinal mucosa and its link to Helicobacter pylori (H. pylori) micro-organisms.Forty patients (30-67 years of age) attending Endoscopy Unit of TBRI were enrolled. They were classified into two main groups; group (I): included 10 dyspeptic patients with no history of acid suppressing drug intake, group (II): included 30 patients complaining of dyspepsia or heart burn receiving proton pump inhibitor (Omeprazole) orally in a dose of 20 mg/day for 1-6 months [Benzimidazole = 5-Methoxy-2-(4-Methoxy-3,5-diMethyl-2-pyridyl-methylsulphinyl)] (C17H19 N3O3S= 345.4) as a treatment for gastro-esophageal reflux disease (GERD) or peptic ulcer.All cases were subjected to routine clinical examination, laboratory investigations and upper gastrointestinal endoscopy. Gastric and duodenal biopsies as well as duodenal aspirate were obtained from all cases. All biopsies were assessed using haematoxylin/eosin and Masson trichrome stains for histopathological evaluation, Geimsa stain for H. pylori detection and immunohistochemistry for proper identification of G. lamblia trophozoites.There was a clear male predominance in Giardia-positive patients. They tended to be older than control cases. Diarrhea, early satiety, abdominal pain or bloating were encountered more in positive cases. In acid suppressed patients (group II), giardiasis was detected in 40% of gastric biopsies, combined with H. pylori in 16.67%. Meanwhile; G. lamblia was evident in 76.67% of duodenal biopsies and 43.33% of duodenal aspirate. In gastric giardiasis, lymphocytic gastritis and intestinal

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metaplasia were detected in 75% and 58.33% of cases respectively and were predominated in combined infection (100%). Duodenal giardiasis caused inflammatory infiltrate of the lamina propria in all cases (100%) with flattening and shortening of duodenal villi in 86.95% . Our results conclude that: Gastric acid suppression encourages G. lamblia colonization in the stomach manifested by lymphocytic gastritis or intestinal metaplasia. Gastric pathogenicity is more evident in combined G. lamblia and H. pylori infections. Duodenal biopsy is more accurate than duodenal aspirate in detecting G. lamblia infection. Caution in prolonged use of gastric acid suppressive drugs should be adopted in G. lamblia endemic areas. PP4-280 RHEUMATOID ARTHRITIS ASSOCIATED WITH SPLENIC LEISHMANIASIS Ioannis Venizelos, Zoı Tatsiou, Anastasios Chatzitolios, Alexandra Moulla Department of Pathology, Hippokration Hospital, Thessaloniki, Greece Rheumatoid arthritis (RA) is a chronic immune-complex disease which affects mostly women during the second and third decade of life. Due to prolong treatment with immunosuppressive drugs and the immunodeficiency which develops later in the course of the disease, it has rarely been described development of malignancies and opportunistic infections. Splenic leishmaniasis after treatment for RA has not been reported. We report the case of a 65-year-old woman who was admitted in our hospital with fever, night sweats, abdominal pain for the last 2 weeks, and weight loss (10 kgr) during the last 3 months. From her past medical history the patient was receiving treatment with methotrexate due to RA for the last 5 years. On physical examination it was found a palpable spleen, about 20 cm below the left costal margin. No peripheral lymphadenopathy, hepatomegaly or skin rushes was found. Hematological examination revealed pancytopenia. Because of the huge size of the spleen and the pancytopenia, a splenectomy was performed. Histological examination of the excised spleen revealed macrophages stuffed with ingested parasites, morphologically consistent with leishmania. The patient had no history of traveling to endemic areas with leishmaniasis. Test for leishmania using indirect immunofluorescence was strongly positive. A bone marrow biopsy revealed leishmania parasites in macrophages. The patient received treatment with amphotericine B and 3 years later she is in an excellent condition with the hematological abnormalities been resolved. In the present study we report the first case of splenic leishmaniasis after treatment for RA. Among other opportunistic infections, visceral leishmaniasis should be suspected after prolong treatment for RA. PP4-281 SCHISTOSOMA HAEMATOBIUM (EGYPTIAN STRAIN): RATE OF DEVELOPMENT AND EFFECT OF PRAZIQUANTEL TREATMENT Sanaa Botros, Olfat Hammam, Naglaa El Lakkany, Said Saif El Din, Fatma Ebid Departments of Pharmacology and Pathology & Theodor Bilharz Research Institute,Warrak El-Hadar, Imbaba, Giza, Egypt This study investigates the development of Egyptian strain of Schistosoma haematobium (S. haematobium) and the resultant immunohistopathological and biochemical changes of organs likely affected. Meanwhile the response of different developmental stages of S. haematobium worms to praziquantel (PZQ) was examined. S. haematobium-infected hamsters (400 cercariae/hamster) were classified into 4 main groups, they were treated 35, 55, 75 and 95 days post infection (PI). Each group was subdivided into three subgroups; two of them were treated orally with PZQ in a dose of 300 mg/kg or 500 mg/kg divided on two consecutive days, while the third one was left without treatment.

Treated groups were sacrificed 20 days post treatment. Infection with S. haematobium became patent 75 days PI, tissue egg load and worm fecundity were higher 95 days and maximal 115 days PI with oogram pattern comparable to that in S. mansoni infection. In the liver, small cellular granulomas were observed 75 days PI with preponderance of CD4+ T-cell phenotypes. In the urinary bladder, only sub-mucosal focal Brunn's nests formation and angiogenesis without typical granulomas were observed. 95 and 115 days PI, confluent granulomata with multiple eggs in the center were observed in the liver and urinary bladder with preponderance of CD8+ positive T cells in the liver and hyperplasia of the urinary bladder lining epithelium with cystitis cystica and papillae formation. Higher worm eradication was recorded with the higher dose of PZQ tested in animals treated 75 and 95 days PI. In conclusion, in spite of the long prepatent period of the Egyptian strain of S. haematobium, sensitivity to PZQ was recorded early after infection. Granulomata were similar to that of S. mansoni in the livers and urinary bladders yet they were confluent with multiple eggs in the center, hyperplasia of the urinary bladder urothelium with cystitis cystica, papillae and Brunn's nests formation predictive of malignant changes with no hepatocyte dysplasia were detected. PP4-282 A DIAGNOSTIC PITFALL IN SAMPLES FROM BRONCHIAL ASPIRATES: A FALSE HUMAN CONTAMINATION CAUSED BY AUREOBASIDIUM PULLULANS IN THREE PATIENTS Hofman Paul1, Butori Catherine1, Le Fichoux Yves2, Hofman Paul1 1 Laboratory of Clinical and Experimental Pathology, University of Nice, Nice, France 2 Laboratory of Mycologie, University of Nice, Nice, France Background: A. pullulans is a saprophyte dematiaceous fungi, which can exceptionnally caused infection in human. Peritonitis, cutaneous infection, meningitis, or keratitis, abcesses in the spleen and jaw due to A. pullulans have been described in adults. In the lower respiratory tract, Aureobasidium species have been very exceptionally isolated. Patients and results: Three cases were observed in a single laboratory, the same week. The patients were: a 60-year-old man presented with a left upper lobe tumor and carcinomatous cells in bronchial smears; a 39-year-old woman with pneumonia and numerous inflammatory cells in bronchial smears; and a 67-year-old man with chronic cough and normal epithelial ciliated cells in bronchial aspirates. In all smears, numerous yeasts measuring from 3 microns to 5 microns were observed. In 2 cases, ellipsoidal unicellular smooth conidia, variable in size, sometimes budding, and originating from rare septate hyphae were noted. These fungi were stained with Papanicolaou, May Grunwald Giemsa, Alcian blue, mucicarmin, and Gomori-Grocott methods. The same fungi were isolated from saponin solution used for red cells lysis before smear staining. Diagnosis was made by comparing the morphology of fungi, both in bronchial smears and in saponin. Finally, fungal culture from saponin isolated A. pullulans. It was well identified due to the presence of large thick-walled black cells that have small single-celled conidia budding. Comments and conclusion: exogenous contamination by microorganisms can frequently caused false diagnosis of infectious diseases in pathology. However, these contaminations should be suspected when the same features are observed in different samples from the same laboratory, when the morphology is very unusual, and/or when non inflammatory cells are detected. A number of fungi of low virulence, including A. pullulans are often considered contaminants when isolated from healthy hosts. However, a recent review of the English literature revealed 25 clinical cases of Aureobasidium spp. infection. Moreover, A. pullulans has affinity for synthetic materials and surgically implanted Silastic devices, causing infection in human and making the differential diagnosis with false contamination very difficult.

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PP4-283 PATOGENETIC VARIANTS OF CASEOUS PNEUMONIA FROM THE POINTOF VIEW OF TEACHING ABOUT PATHOMORPHISM OF TUBERCULOSIS Eugene Kazachkov, Polina Tselischeva State Medical Academy, Chelyabinsk, Russia The aim of this research is to study the structure peculiarities of pathogenetic variants of caseous pneumonia (primary and secondary) in the 80th years of XX century and present days. Material and methods. The comparative patomorphologic analysis of the deaths cases of patients having tuberculosis of breathing organs based on the materials of specialized prosectorum of Chelyabinsk regional antituberculosis dispensary for 1986 (99 observations, 1st group) and 2006 years (105 dead patients, 2nd group) were held. Results. Was found the increasing of widespread destructive forms of tuberculosis infection such as fibrocavernous tuberculosis of the lungs (61%) and primary caseous pneumonia (PCP, independent clinic anatomic form of tuberculosis; 20%). The basic structural display of acute progressiveness of tuberculosis infection of the lungs is caseous pneumonia which can be as independent form of tuberculosis (PCP) and also it can be the stage of progress of another clinic morphologic forms of this infection (secondary caseous pneumonia: SCP). In 1986 PCP was registered in 3 cases (about 3%). The process was unilateral with the lesions of the higher lobe of the lungs. In 2006 PCP was founded in 20% of observations of tuberculosis of breathing organs. In the second group was a registered bilateral lesion of the lungs with the localization of the process more than in 3 lobes. Histological research showed that separating the zone of caseous necrosis polymorphocellular torus was introduced by lymphocytes, macrophages, neutrophils, few epithelioidcells. The density of demarcating cellular infiltrate in the observation of 2nd group was significantly lower than that of the 1st group. SCP developing in the terminal stage of another forms of tuberculosis: infiltrate, acute cavernous, fibrocavernous. The growth of the frequency of SCP with 23.2% in the 1st group, up to 68.6% in the 2nd group. In the most cases SCP was stipulated by progressiveness of fibrocavernous tuberculosis (60.8% in 1986 and 86% in 2006). Conclusion. For the last time the growth of observations with caseous pneumonia is registered. One of the structural peculiarities of modern pulmonary tuberculosis in the development of the (SCP) which in the most cases complicates the course of fibrocavernous tuberculosis. This situation from the point of the teaching about pathomorphism may be interpreted as a negative stage of pathomorphism of tuberculosis. PP4-284 CYSTIC ECHINOCOCCOSIS OF THE THYROID GLAND: A CASE REPORT AND DIAGNOSTIC PROBLEMS Kemal Behzatoglu, Pelin Yıldız, Sule Canberk, Meltem Oznur, Gulben Erdem Huq, Erol Rustu Bozkurt Istanbul Educational and Research Hospital, Turkey In humans, cystic echinococcosis is one of the most serious helminthic diseases worldwide. The larval stages (cysts) of Echinococcus granulosis are mainly located in liver and lung. Other locations of infection are rare, which may occasionally hinder its diagnosis. A 65-year-old woman was admitted with a swelling in the right lobe of the thyroid gland of 2 years duration. Laboratory investigations and thyroid function were normal. Thyroid scanning showed a cold nodule in the lower pole of the right thyroid lobe. Thyroid ultrasonography showed multiple cystic nodules in the thyroid gland. To maka a differential diagnosis of the thyroid nodule, fine-needle aspiration biopsy (FNA) was performed and diagnosed as nonspesific inflammation. During aspiration biopsy, the patient did not present a clinical picture of anaphylactic reaction. Subtotal thyroidectomy was carried out and histopathologic examnation

revealed a hydatid cyst. Subsequent to the histologic diagnosis (which was hydadit cyst), review of the FNA biopsy showed acellular laminated fragments; it had not been taken into consideration before. In endemic regions-like our country, patients presenting with cystic nodular lesions of thyroid-unusual location of hydatidosis- hydatic cyst should be keept in mind. PP4-285 GASTRIC PRESENTATION OF STRONGYLOIDIASIS IN A DIABETIC PATIENT Safak Ersoz, Havvanur Turgutalp, Remzi Akdogan, Ismail Saygın, Umit Cobanoglu, Sevdegul Mungan, Orhan Ozgur Karadeniz Technical University Medical Faculty,Trabzon,Turkey Strongyloides stercoralis is a common intestinal helminth. Strongyloidiasis is a worldwide parasitic infection affecting approximately 75 million people. Prevalence of up to 25% has been reported. In healthy hosts, the parasite usually does not cause any symptoms, or only mild symptoms those are limited to the small intestine. In immunocompromised host, massive infection may occur, that is causing hyperinfection syndrome or disseminated strongyloidiasis. There are few reports of gastric involvement, especially presenting as pylor stenosis. A 78 year-old male patient had admitted to local city hospital with complaints of epigastric pain, nausea and vomiting. He had those complaints since two months. An upper gastrointestinal endoscopy had revealed hyperemia and erosions at gastric antrum. The patient had been discharged after a twenty days treatment period with relieve of the symptoms. After a short period the same symptoms showed a recurrance and he was referred to our university hospital. Meanwhile he had a weight loss of 25 kg. At his admittance, physical examination was almost normal other than peripheral edema. Laboratory examination revealed a hyperglicemia of 161 mg/dl and hypokalemia with hypochloremia. An upper gastrointestinal endoscopic examination was performed again. In this procedure eosophagitis at the lower segment of eosophagus was observed. Cardia and fundus were normal, while edema was observed at corpus and antrum with pylor stenosis. In histopathological examination of gastric biopsy, adult forms of Strongyloides stercoralis were found in gland lumen and lamina propria accompained with mixed inflamatory cell infiltration and regenerative changes at gastric epithelium. As he refused the therapy, the patient did not receive a spesific treatment for strongyloidiasis. Here we report a case with diabetes mellitus and gastric strongyloidiasis leading to pylor stenosis. PP4-286 UNCOMMON LOCATIONS OF HYDATID CYST Nebil Bal1, N. Emrah Kocer1, Rabia Arpaci2, Ali Ezer3, Fazilet Kayaselcuk1 1 Baskent University Medical Faculty Department of Pathology, Turkey 2 Tuncer Pathology Laboratory Adana, Turkey 3 Baskent University Medical Faculty Department of General Surgery, Turkey Hydatid cyst is a parasitic disease, formed by Echinococcus granulosus, which is a social and economic problem in developing countries that effects public health directly. Liver and lungs are the most common locations of hydatid cyst but it may develop in any part of the body. In this study we analyzed 154 cyst hydatid and presented cases with uncommon locations as spleen, bone, intraarterial, ovary, adrenal, heart, mesenteric, retroperitoneal, subcutaneous tissue, breast, intramuscular tissue. We concluded that the diagosis of cyst hydatic should be considered in patient with cystic mass if the patient lives in geographic region that have a high risk for Echinococcus granulosus or migrated from or visited an endemic area.

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PP4-287 THROMBOTIC THROMBOCYTOPENIC PURPURA (MOSCHOWITZ SYNDROME) AND DEATH FOLLOWING YELLOW FEVER VACCINATION Angela Fıor1, Annie Motard2, Philippe Dussart3, Félix Djossou4 1 Service d’Anatomie et Cytologie Pathologique, Centre Hospitalier de Cayenne, Cayenne Cedex, French Guiana 2 Laboratoire de Biologie Polyvalent, Centre Hospitalier de Cayenne, 3 Laboratoire de Virologie, Institut Pasteur de 1 la Guyane, Cayenne, French Guiana 4 Unité Médicale des Maladies Infectieuses et Tropicales, Centre Hospitalier de Cayenne, Cayenne Cedex, French Guiana A 40-year-old woman had, received yellow fever vaccination before her travel in French Guiana, presented after 10 days at emergency room of Regional Hospital of Cayenne (French Guiana) with complaints of fever and general malaise since 3 days. On examination, she has arthralgia, arm and leg ecchymosis and petechia, paleness mucosa, headache and discrete confusion. Laboratory test showed an haemolytic anaemia, an thrombopenia in favour a thrombotic thrombocytopenic purpura (TTP)(Moschowitz Syndrom). Evolution go to worse at death the 09 January 2006 and autopsy was performed revealed widespread microthrombosis of small vessels in all the examined organs in conformity with suspected TTP. The onset of symptoms soon after vaccination and failure to identify any other cause suggest that the TTP was an adverse reaction to the yellow fever vaccine. PP4-305 SHOCK SYNDROME AND ADVERSE OUTCOME OF A CASE OF DENGUE FEVER CAUSED BY DEN-2 VIRUS INAUGURAL OF 2006 EPIDEMIC DISEASES IN FRENCH GUIANA Angela Fior1, Philippe Dussart2, Abdelfatteh Zeddini1, Félix Djossou3 1 Service d’Anatomie et Cytologie Pathologique, Centre Hospitalier de Cayenne, Cayenne Cedex, French Guiana 2 Laboratoire de Virologie, Institut Pasteur de la Guyane, B. P. 6010, Cayenne, French Guiana 3 Unité Médicale des Maladies Infectieuses et Tropicales, Centre Hospitalier de Cayenne, Cayenne Cedex, French Guiana If in neighbouring states, French Guiana is regularly affected all 2 at 6 years for a Dengue fever epidemiological crisis because local conditions are more favourable at Aedes aegypti development. The four serotypes of Dengue fever are present in French Guiana but DEN-2 was more predominant in 2006 strong epidemic. This affected more than 13 700 subjects and caused four death directly release with DEN-2. In this epidemic, our cases was the second death, a four years old child, death at home on night 30-31 March. He consult tree days ago and present any sign of gravity. This case take all measures to confirm aggressively of serotype and more then 180 diseased was recovered with 57% by strong non hemorrhagic form. Blood samples not obtained, the aim of post mortem investigations was taking tissues and liquids for Dengue fever confirmation from Pasteur Institute of Guiana. These demonstrate a typical hypovolemic Dengue shock syndrome with poorly hepatic hemorrhagic lesions but diffuses acute histological alterations of sweat acini’s.

Miscellaneous PP4-288 INTRODUCING PATHONET, A VIRTUAL HISTOLOGY LABORATORY László Fónyad1, Attila Zalatnai2, Béla Molnár3, László Kopper2 1 Department of Pathology, Semmelweis University, 3DHISTECH Ltd., Budapest, Hungary 2 Department of Pathology, Semmelweis University, Hungary 3 Department of Internal Medicine, Semmelweis University, 3DHISTECH Ltd., Budapest , Hungary

BACKGROUND Virtual microscopy has become accepted, both as a new research tool and as an application used in education and in routine surgical pathology as well. After the problem of high-quality, whole-slide-digitizing was solved we are working to built up a holistic virtual histology laboratory. AIMS During the past few years the staff of the 1. Dept. of Pathology, Semmelweis University, in association with 3DHISTECH Ltd. were continuously developing a web portal, mainly for pathologists, aiming to provide a platform, to share and consultate problematic cases, and to built up an open access knowledge pool, where interesting cases are to be discussed and virtual slide seminars are organized. As an University institute we also emphasize to provide easy-to-handle and up-to-date web-based digital reference book for our students, thus we uploaded both undergraduate and postgraduate educational materials, for medics and for pathology residents as well. The aim of this presentation is to acquaint the features of this virtual meeting place: Pathonet RESULTS Till the submission of this abstract, about a hundred users have registered on the portal, we had about 9000 page loads from abut 900 unique and 500 returning visitors so far which is a doubling in the number of visitors in a year. Till 2006, the portal guarantees the background for the Hungarian Slide Seminars. Previewing and the discussion of the cases in our institute is no longer made with optical, but the virtual slides. Preparing for exams the students can use another feature on the portal. In an e-book form the uploaded slides of the educational material will be completed with texts, macroimages and comments. From the beginning of the next semester the whole concept of histology teaching will be changed in our Institute. Instead of the optical microscopes we use computers and via the portal the students could revise the slides after practices and before exams. Recently we have developed a new, completely free feature for our registered users on our portal for editing cases and uploading whole slides on Pathonet server to use it as reference to their publications. The server generates a unique link to the slide. Installing the MiraxViewer and typing the link to a webbrowser or clicking on it if it appears on the online form of a journal article, the slide will open in a MiraxViewer window. CONCLUSIONS Pathonet portal is a user-friendly meeting place for pathologist. PP4-289 “I’AM STILL LEARNING” – INTRODUCING E-SCHOOL, A NEW WAY TO STUDY PATHOLOGY László Fónyad1, Attila Zalatnai2, Levente Ficsór3, Máté Montvai3, Béla Molnár4, László Kopper2 1 Department of Pathology, Semmelweis University, 3DHISTECH Ltd. , Budapest, Hungary 2 Department of Pathology, Semmelweis University, Hungary 3 3DHISTECH Ltd. , Budapest, Hungary 4 Department of Internal Medicine, Semmelweis University, 3DHISTECH Ltd. , Budapest, Hungary

BACKGROUND During the past years the developments on the field of virtual microscopy have reached a professionally accepted higher level. Now the question is no longer weather the digital microscope could be applied equivalently to the conventional microscopes but what additional benefits can we gain using the digital slides. One of the most popular field of using digital slides is education. As a University Institute,

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teaching pathology, we also encounter the most serious challenges, measuring up to the expectations of our prime privileged customers, the students. AIMS On the 8th European Congress on Telepathology and 2nd International Congress on Virtual Microscopy we reported our first experiences on building up our telepathological workstation and testing it for gradual education. After the success of virtual microscopy on pilot histology practices we decided to replace the conventional microscopes to computers and in strong cooperation with 3DHISTECH Ltd. to design and create an educational software: E-School. RESULTS The E-School software-package now contains 3 major parts: an encoder, a viewer and an E-School server software. With the encoder you can encrypt your document, written in html format, so Universities or publishers of any kind can protect their materials and can ask license fee for them. Of course these documents can be free as well. The viewer can decode the documents and in case of non-free materials, via the E-School server the product keys could be handled by the product owners. The major advantage of the system is that digital slides, stored on a slide server or on DVD can be linked to the document allowing entire slide access. There is a self-testing and an exam module in the software-package as well. The educational material of our Institute is now fully digitized and a histology textbook is written with the E-School softwer. On the beginning of the next semester we launch our digital histology lab in regular practice. CONCLUSIONS In the era of e-learning it is outmost important for Univerities to provide a streamlined platform for her students where they can benefit all the possibilities of the advances in computer sciences. The E-School system is a good solution for compiling educational material for students and as the documents could be encrypted as well, it is also appropriate for publishing e-books of histology with real rich-in-details images and entire slide access. PP4-291 AUTOMATED FLUORESCENT SLIDE SCANNING Viktor Sebestyén Varga1, Levente Ficsor2, Viktor Kamarás1, Béla Molnár2, Zsolt Tulassay2 13DHISTECH Ltd., Hungary 2 Second Department of Medicine, Semmelweis University, Budapest, Hungary

Introductions: Today there are several whole slide imaging systems for bright field, but there is no product supporting fluorescence scanning. To digitize the daily workload of a routine histology laboratory walk away automation is needed. Our goal was to develop a system which is capable of scanning both bright field and fluorescent samples automatically. Materials: We used a Mirax Scan (Carl Zeiss MicroImaging, Thornwood, NY) whole slide imager with an optional filter changer with fluorescent filters for DAPI, FITC and Rhodamine. The light source was an X-Cite 120 metal halide lamp (EXFO, Vanier, QC). The Marlin F-146C (Allied Vision Technologies, Stadtroda, Germany) fire wire camera was applied for image capture. A Celsius PC (Fujitsu Siemens Computers, Maarssen, Netherlands) with dual 3.2 GHz CPU and 3 GB RAM was used for controlling the scanner microscope. We developed the software with Visual Studio (Microsoft, Redmond, WA) and C++ Builder (Borland Software, Scotts Valley, CA). Methods: Mirax Scan uses a preview camera to locate tissue samples on a slide. Fluorescent samples are usually invisible to such a low magnification camera in normal illumination. To overcomes this issue users are required to mark the sample with a marker pen. At fluorescent slides the exposure times can vary from few milliseconds to several seconds and the different channels have different exposure times. In the selected area Mirax Scan is auto focusing on grid points using a special algorithm developed for fluorescence, which is continuously adjusting the exposure time. On the focused field of views exposure times are measured for every selected fluorescent channel. The sample is then digitized using an interpolated focus map and the shortest exposure time in

every channel respectively. Results: A 10 x 10 mm sample using a Plan-Apochromat 20x / 0.8NA objective with a resolution of 0.23 μm / pixel in 3 channels, with exposure times of 52, 100 and 48 ms was scanned in 40 minutes. The system was tested with success in a walk away scanning mode with 30 slides yielding focused images and slides. Field of view black and white image compensation was not necessary. Conclusions: Fully automated multichannel, fluorescent scanning is feasible. Longer scanning times compared to bright field are inevitable but acceptable since the required scanning volume is much lower on average. With further algorithm development and the use of cameras suited for fluorescence scanning can be sped up. PP4-292 PARAGOMINUS WESTERMANI: CASE REPORT Euridice Robles Perez1, Ericka Sagrario Peña Mirabal2 1 Instituto Nacional de Enfermedades Respiratorias, Mexico 2 Instituto Nacinal de Enfermedades Respiratorias, Mexico

Male patient, 80 years old, history of pulmonary tuberculosis (class III, ATS) and diabetes type II. The patient started three days before hospital admission with cough and dyspnea. The day of his admission he presented fever and acute respiratory failure. Arterial blood gas analysis showed respiratory acidosis and blood sample with eosinophilia. He required orotracheal intubations and mechanical ventilatory support. Chest x-ray showed pulmonary bilateral reticulonodular infiltrates and bronchiectasis. The patient died 24 hours after hospital admission. At the post-mortem examination we found histological changes due to diabetes mellitus and tuberculosis. Lung tissue showed bronchiectasis with colonization by aspergillus sp. And paragominus westermani Paragonimiasis is a parasite disease with low frequency of presentation in Mexico, is acquired by eating row crab meat. The body sites affected are lungs and brain. Differential diagnosis is with tuberculosis and fasciolosis. Treatment is albendazol. PP4-293 ADRENAL ADENOMATOID TUMOR Liapis George1, Evangelou Kostas1, Alexandrou Paraskevi1, Felekouras Evangelos2, Delladetsima Ioanna1 1 Pathology Department Laiko General Hospital University of Athens, Greece 2 1st Department of Surgery University of Athens Greece

Adenomatoid tumors are benign neoplasms of mesothelial origin more commonly observed in the paratesticular area of the testis and corpus uterus. Extragenital adenomatoid tumors have also been described in unusual sites such as heart, mesentery, pleura, lymph nodes and adrenal glands. We report on a case of adrenal adenomatoid tumor (AAT) incidentally discovered in a thirty two year-old woman. A surgical operation was performed. Grossly, a greyish-white, solid tumor measuring 6.5 x 5.2 x 3.5 cm occupied almost the entire adrenal gland while only a thin rim of adrenal tissue was preserved. Microscopically, the tumor was composed of anastomosing tubules lined by flat or cuboidal cells with scanty or moderate eosinophilic cytoplasm. An ensuing intricate tubular network resembled hemangioma. Cells with prominent cytoplasmic vacuoles as well as spindle cells were also present. The intervening stroma was scanty and was composed focally of smooth muscle fascicles. At the periphery of the tumor collections of lymphocytes with occasional follicle formation were seen. Immunohistochemically the neoplastic cells were strongly positive for calretinin, cytokeratin 5/6 and WT1, whereas they were negative for HMB-45, SMA, CD34, estrogen and progesterone receptors. In the reviewed literature 29 cases of AAT have been reported, all but one referring to men. As far as histogenesis is concerned, in our case there are no indications of hormonal influence on tumor progression while the development of the tumor within the adrenal gland may be attributed to the common mesodermal origin.

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PP4-294 COMPARATIVE STUDY BETWEEN SEMI-AUTOMATED AND MANUAL CELL QUANTIFICATION IN DIGITAL IMAGES IN PATHOLOGY Marylène Lejeune1, Joaquín Jaen1, Carlos López1, Patricia Escrivà1, Maria Teresa Salvadó1, Ramón Bosch1, Lluís Pons1, Jordi Baucells2, Xavier Cugat2, Jordi Roig2, Tomás Alvaro1 1 Verge de la Cinta Hospital, Pathology Department, Spain 2 Verge de la Cinta Hospital, Informatics Department, Spain Background: Manual quantification of immunohistochemical stained cells is the most frequent method in the current practice in pathology. To avoid the manual subjectivity, a number of automated and semi-automated processes have been previously developed and described. In the present study, we have developed different semi-automated processes in order to quantify different immunohistochemical markers. To obtain a gold-standard reference, we have performed manual quantifications and we have also evaluated inter and intra-observer variability. Method: 196 digital color images were obtained from sections of Hodgkin’s lymphoma biopsies and stained with various nuclear, cytoplasmatic and membrane markers. Each image was manually quantified twice by 3 different observers. To analyse the images, we developed a specific macro for each marker with a commercial analysis software. The global dataset obtained with the software, were dropped to an Excel Datasheet, where the number of cells was obtained after we introduced different corrector factors. To evaluate the agreement degree between manual and semi-automated method and to quantify our intra and inter-observers variability, a comparative statistical analysis was performed with SPSS 11.0. Results: Globally, the variability was higher for intra-observers than for inter-observers counts, except in nuclear stains were both types of variations have similar differences. The comparison between semi-automated and manual methods shows that both methods have the same way of cell quantification. Both manual and automatic quantification showed a higher variation when the images contained more than 100 positive nuclei. Discussion: This study quantifies variation observed in manual quantification and shows that the different semi-automated procedures developed for each marker in our laboratory represent a valid alternative to manual quantification. (PI 04/1440, 04/1467, 05/1527). PP4-295 NEW APPROACH FOR AUTOMATIC QUANTIFICATION OF IMMUNOHISTOCHEMICAL NUCLEAR MARKERS IN DIGITAL IMAGES Joaquín Jaén1, Carlos López1, Marylène Lejeune1, Patricia Escrivà1, Maria Teresa Salvadó1, Ramón Bosch1, Lluís Pons1, Jordi Baucells2, Xavier Cugat2, Jordi Roig2, Tomás Alvaro1 1 Verge de la Cinta Hospital, Pathology Department, Spain 2 Verge de la Cinta Hospital, Informatics Department, Spain Background: Several procedures of digital image analysis have been developed for quantitative evaluation of nuclear immunohistochemical markers with prognostic, diagnostic and therapeutic significance in current medical practice. To our knowledge, do not exist effective algorithms that permit to obtain a precise cell quantification in digital images with a high grade of complexity. The aim of this study is to develop a new methodology capable to analyze correctly, images with high variability in their morphology, cellular density, stain intensity and cell distribution. Method: 118 digital images from 4 different immunohistochemical nuclear markers were captured with different grade of cell concentration and clusters composition. Two coordinated macros were elaborated to perform the automatic count of positive nuclei. The first macro was developed with a commercial software that allows the modification and segmentation of the images. All extracted

information was dropped to an Excel datasheet, where we have developed a macro with Visual Basic and introduced different algorithms that manage the dataset obtaining a final number of positive nuclei on each image. All statistical analysis was performed with SPSS 11.0. Results: t-Student test, Spearman correlation and ICC, showed no significant differences between the manual and the automatic count, whatever the image complexity. Kaplan-Meier and Bland Altman graphic representations indicate that cluster composition and a high nuclei density, increase variability on both types of count, and globally in more than 90% of images, automatic count was similar to the manual. Conclusion: This study, describes a new methodology for the automatic count of digital images for different immunohistochemical nuclear markers. This method improves the quantification of images with a high complexity in their nuclear composition. Variability observed between human and automatic count are the same as inter or intra-observer variability, which is accepted in the current clinical practice. (PI 04/1440, 04/1467, 05/1527). PP4-296 EFFECTS OF DIGITAL IMAGE COMPRESSION ON COMPUTER-ASSISTED IMAGE QUANTIFICATION OF IMMUNOHISTOCHEMICAL STAINED CELL NUCLEI Carlos López1, Joaquín Jaén1, Marylene Lejeune1, Patricia Escrivà1, Maria Teresa Salvadó1, Ramón Bosch1, Lluís Pons1, Jordi Baucells2, Xavier Cugat2, Jordi Roig2, Tomás Alvaro1 1 Verge de la Cinta Hospital, Pathology Department, Spain 2 Verge de la Cinta Hospital, Informatics Department, Spain Background: The analysis of digital images and standard compression algorithms in current clinical practice has been used since some years ago. Image compression may reduce the amount of computer memory required for store images. The consequences of image compression have been previously evaluated on different diagnostic techniques. The aim of this study is to analyse the consequences of computer-assisted quantification, between uncompressed Tiff format and different levels of image compression in Jpeg format of immunohistochemical stained cells. Method: Digital images were captured with the software Leica IM50 4.0, in Tiff format from tissue samples stained with immunohistochemical markers, Ki67 (n=24) and FOXP3 (n=24). In a second step Tiff images were converted with the software ACDSee 6.0 to Jpeg files with a compression factor of 0, 50 and 100%. All the captured and converted images (n=196), were analysed with two coordinated macros developed with an image analysis software and an Excel datasheet. Quantified parameters were the total stained positive area and the number of positive nuclei. Tiff images results were compared with the different compression factor in Jpeg format, with Kaplan-Meier and Bland Altman plots using SPSS 11.5 Statistical Software. Results: Globally, variations in the quantification of positive stained areas between Tiff and the different compressed files are similar. On the other hand, cell count differences are lower with a compression factor of 0% and higher with a 50 or 100% of compression. Nevertheless, all compressed formats have a similar variability when images have less than 100 nuclei, and in this case, this variability is lower than globally. Conclusion: This descriptive study shows that image compression is a source of variation in automatic count of digital images. In these conditions, image compression should be omitted for immunohistochemical digital image quantification, overall in images with more than 100 nuclei. (PI 04/1440, 04/1467, 05/1527)

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PP4-298 MUCOEPIDERMOID PAPILLOMA OF CONJUNCTIVA Cem Comunoglu1, Ahmet Midi2, Akin Banaz3, Onder Peker2 1 Oruc Pathology Laboratory, Turkey 2 Maltepe University, School of Medicine, Department of Pathology, Turkey 3 Dunya Goz Hospital, Department of Eye Diseases, Turkey Background: Mucoepidermoid papilloma of conjunctiva is a papillary lesion. Columnar cells and Goblet cells line these papillary structures. Exophytic mucoepidermoid papilloma of conjunctiva has not been reported in the literature previously. Case report: Forty-three year-old male patient presented with a cojunctival lesion. Excisional biopsy was performed. Macroscopically the greatest dimension of the tumor measured 0,9 cm. Microscopically, a papillary lesion was seen. Lining epithelium was columnar cells and Goblet cells. A diagnosis of mucoepidermoid papilloma of conjunctiva was given. Conclusion: Conjunctival mucoepidermoid carcinomas have been described but there is no evidence of the presence of any precursor lesions. Two separate case reports describe inverted mucoepidermoid papillomas. Present case had a papillary configuration and did not contain an inverted component. Although conjunctival papillomas show HPV effects in present case cytopathic HPV effects could not be detected. It can be predicted that these lesions do not carry a potential of malignancy but because there is no previous information about biological behavior potential of these lesions, strict clinical follow-up is recommended. Whether mucoepidermoid papilloma is a precursor lesion of mucoepidermoid carcinoma of conjunctiva is a problematic issue which should be solved in future reports. PP4-299 OXALATE CRYSTAL DEPOSITION ASSOCIATED WITH MULTIPLE INFARCTION IN THE SPLEEN OF A PATIENT WITH NON-CIRRHOTIC PORTAL HYPERTENSION Engin Cigerciogullari1, Suha Goksel1, Abdullah Sonsuz2, Hasan Tasci3 1 Department of Pathology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey 2 Department of Gastroenterolgy and Hepatology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey 3 Department of General Surgery, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey Splenic infarction is an unusual finding of cirrhotic or non-cirrhotic portal hypertension. Dystrophic oxalosis can be seen in thyroid, breast, and retina in many conditions. Oxalate deposition is well known in renal pathology, and had been reported in many organs in patients with primary and secondary oxalosis. We couldn’t find oxalate deposition in the spleen in the literature. The presented case was a 59-year-old man with non-cirrhotic portal hypertension. Pathological examination of splenectomy revealed moderate splenomegali, multifocal infarctoid areas causing capsular retraction. Microscopic investigation revealed multifocal infarction of different ages. Gandy-gamma body formations were also present. Surprisingly, multifocal heavy oxalate crystal depositions around the necrotic area, and perivacular spaces in vessels nearest to infarction were seen. With the presentation of this case, spleen can be included in the oxalate depositing organs. The oxalate deposition in the spleen may be dystrophic due to infarction, or may also be the first sign of hyperoxaluria.

PP4-300 A CASE OF PARARENAL RETOPERITONEAL CASTLEMAN DISEASE: INCIDENTALY DETECTED AS AN ADRENAL GLAND MASS Evrim Kus1, Yesim Gurbuz1, Cengiz Ercin1, Nagihan Inan2, Ozdal Dilloglugil3 1 Kocaeli University Medical Faculty Pathology Department, Turkey 2 Kocaeli University Medical Faculty Radiology Department, Turkey 3 Kocaeli University Medical Faculty Urology Department,Turkey INTRODUCTION: Castleman’s disease, or angiofollicular lymph node hyperplasia, is a relatively rare disorder characterized by benign proliferation of lymphoid tissue. Castleman’s disease is usually related to chronic herpes virus infection and it is usually localized in the mediastinum. Retroperitoneal and especially pararenal localization is very rare. CASE REPORT: A retroperitoneal mass was incidentally found on the abdominal ultrasonographic imaging of a 51 year-old woman. Surgery was planned with a preoperative diagnosis of non-functioning adrenal gland neoplasia. The tumor was removed by retroperitoneal laparoscopic adrenalectomy. In gross examination the tumor was an encapsulated, homogenous pale colored mass measuring 4.0x5.5x5.5 cm. Microscopic examination showed a histologically normal adrenal gland, and a large lymph node rich in vascular structures. The nodal architecture was altered by an increased number of lymphoid follicles. These lymphoid follicles were unusually small and their germinal centers consisted of radially penetrating capillaries, some of which formed a characteristic “lollypop” structure. Many of these capillaries were surrounded by wide cuffs of hyaline substance. Multiple concentric layers of mature lymphocytes were surrounding the small germinal centers. Thus, the tumor was pathologically diagnosed as hyaline-vascular type angiofollicular lymph node hyperplasia (Castleman’s disease) of a peri-adrenal gland lymph node. DISCUSSION: In this poster, we present a case of Castleman’s disease localized in the retroperioneum next to the uninvolved adrenal and the kidney. Although it is an unusual localization Castleman’s disease should be included in the differential diagnosis of retroperitoneal masses. PP4-301 LEPROSY: REPORT OF THREE CASES AND CURRENT SITUATION IN TURKEY Ozgur Mete1, Nesimi Buyukbabani1, Mustafa Sutlas2, Reyhan Uzdil2, Guzin Ozarmagan3 1 Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Turkey 2 Istanbul University, Leprosy and Venereal Diseases Research Center, Turkey 3 Istanbul University, Istanbul Faculty of Medicine, Department of Dermatology, and Leprosy and Venereal Diseases Research Center, Turkey BACKGROUND: Diagnosis of leprosy is most commonly based on the clinical signs and symptoms. Laboratory confirmation of leprosy requires the demonstration of acid-fast bacilli in a skin biopsy. We report herein 3 cases which were diagnosed as borderline lepromatous (BL), lepromatous (LL) and borderline tuberculoid leprosy (BT) last year in our department. Among them, only one case was clinically suspected as leprosy before the histopathological examination. REPORT OF THE CASES: Case 1: A 26-year-old man had been complaining of spontaneously regressing and recurrent erythematous anesthetic macules and infiltrated lesions on his forearms and legs since 2 months. N. radialis and ulnaris were bilaterally palpable. Clinically vasculitis, Sweet syndrome and syphilis were suspected and a punch biopsy was done. The histopathological diagnosis was BL. Case 2: A 56-year-old man had been suffering from erythematous macules and pathches on the whole body and nasal congestion

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since 3 years. N. ulnaris, radialis, poplitealis and auricularis were bilaterally palpable. Clinical diagnosis was LL, which was fully compatible with the histopathological examination of the punch biopsy. Case 3: A 45-year-old man had pale anesthetic macular lesions with erythematous border on his face, arms and back since 4 years. Clinically erythema annulare santrifugum and erythema dischromicum perstans were suspected and a punch biopsy was done. The histopathological diagnosis was BT. CONCLUSION: Leprosy is endemic in tropical and subtropical Asia, Africa, Central and South America and the Pacific regions. In Turkey, 2 595 cases were registered to our leprosy research center. Seven new cases were detected last year. Three of them, which were diagnosed in our department, were presented. As reported to WHO by 115 countries in 2006, 296 499 new cases were detected last year. Clinically, the skin lesions often resemble those of lupus erythematosus, sarcoidosis, syphilis, erythema nodosum, erythema multiforme and cutaneous tuberculosis. The most effective way of preventing disabilities in leprosy, as well as preventing further transmission of the disease, lies in early diagnosis and treatment with multi-drug therapy. PP4-302 ULTRASTRUCTURAL STUDY IN A CASE OF PRIMARY BREAST CARCINOMA WITH OSTEOCLAST-LIKE GIANT CELLS Semen Yesil1, Bulent Ahiskali2, Sitki Tuzlali3, Ekrem Yavuz1, Ridvan Ilhan1 1 Department of Pathology, Istanbul Medical Faculty, University of Istanbul, Turkey 2 Department of Histology and Embryology, Istanbul Medical Faculty, University of Istanbul, Turkey 3 Department of Pathology, Istanbul Medical Faculty, University of Istanbul, Turkey Primary carcinoma with osteoclast-like multinucleated giant cells is a very rare tumor of the breast. We present the light and electronmicroscopic features of a case of invasive ductal carcinoma containing osteoclast-like multinucleated giant cells with regard to the histogenesis (origin) of giant cells. A 48-year-old woman with a 3 cm-mass in the middle part of her right breast was admitted to our hospital, and she underwent modified radical mastectomy. Macroscopically the tumor was well circumscribed with a grey-white cut surface. Microscopically the tumor was invasive ductal carcinoma with cribriform features and many osteoclast-like multinucleated giant cells between the neoplastic glands and within their lumina. Of the eleven axillary lymph nodes dissected, two were metastatic. The lymph-node metastasis also contained osteoclast-like multinucleated giant cells and stromal reactions. On immunohistochemical analysis, multinucleated giant cells were positive for CD68 and alpha-1-antitrypsin and negative for keratins, revealing their histiocytic origin. Neoplastic cells were positive for keratins and estrogen and progesteron receptors, and negative for c-erb B2. Ultrastructural analysis revealed multinucleated cells with many lysosomes, supporting their histiocytic origin, in vicinity of ordinary glandular cells, elsewhere intermixed with them. They contained neither cell junctions nor microvilli. We were able to detect cytoplasmic fusion of some mononuclear cells of the same character. We conclude that osteoclast-like giant cells are formed by fusion of mononuclear histiocytic cells.

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