Aim: The aim of this study was to establish the expression and the significance of angiogenic markers in T1 bladder cancer with concomitant carcinoma “in situ”. Introduction: It has been determined that overexpression of hypoxia-inducible factor 1-alpha (HIF-1alfa), vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor1 (VEGFR1) correlates with tumor grade, disease progression and recurrence, as well as poor overall survival. Carcinoma in situ (CIS) is frequently seen in conjunction with bladder tumors and represents an additional obstacle for management of T1 bladder cancer patients. Material and Methods: The immunohistochemical expressions of HIF-1alfa, VEGF, and VEGFR1 were evaluated in 295 T1 bladder cancer samples, incorporated in tissue microarrays. HIF1-alpha was assessed through nuclear staining, while the angiogenic profile was estimated through cytoplasmatic positivity of the VEGF and VEGFR1. Microvessels were identified by immunostaining of endothelial cells for CD34 and microvessel density (MVD) was presented as the average number of counted microvesels. Results: After a mean followup of 53 months, we found that T1 bladder cancer patients, who had concomitant carcinoma in situ had worse overall survival (p<0.05), furthermore, those tumor samples less expressed HIF-1alfa (p<0.05), and VEGFR1 (p<0.05). Expression of VEGF, VEGFR1, HIF-1alfa, as well as MVD, did not have significant impact to survival rate and further outcome. Conclusion: Worse overall survival and decreased expression for angiogenic markers in samples with accompanying CIS were established. Estimation of HIF-1alfa and VEGFR1 expression emerged as potential diagnostic supplement, selecting the T1 bladder cancer patients that could require an intensive follow-up.

Aim: Here, we explored the role of NCAM/FGFR1 signaling and pro-fibrotic gene expression signatures as initiating or driving forces of EMT program in cultured human proximal tubular epithelial cells. Introduction: Epithelial-to-mesenchymal transition (EMT) contributes to maladaptive repair and parenchymal damage during renal fibrosis. Based on previous reports, neural cell adhesion molecule (NCAM) and fibroblast growth factor receptor 1 (FGFR1) are both considered to be mechanistically involved in the EMT process. Material and Methods: By using an established in vitro model of EMT of the human proximal tubular epithelial cells (HK-2 cells) in response to TGF-β1 (10ng/mL) exposure, NCAM/FGFR1 signaling responses were analyzed by light microscopy, immune-labelling, qRT-PCR and scratch assays. Modulation of FGFR1 was induced using PD173074 (100nM). Distribution of TGF-β1 downstream effectors was assessed in HK-2 cells of the EMT program as well as in 50 biopsies of different human kidney diseases to explore their in vivo correlation. Results: EMT associated with morphological changes started 48h after TGF-ß1 treatment of the HK-2 cells and was clearly apparent after 72 hours, associated with loss of CDH1 (encoding E-Cadherin) and transcriptional induction of SNAI1 (encoding SNAIL), SNAI2 (encoding SLUG), TWIST1, MMP2, MMP9, CDH2 (encoding N-Cadherin), ITGA5 (encoding integrin-α5), ITGB1 (encoding integrin-β1), ACTA2 (encoding α-SMA) and S100A4 (encoding FSP1). After 24 hours of TGF-β1 exposure at the early stage of SPECIJALNA SESIJA: SESIJA SPECIJALIZANATA 7 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. EMT program, transcriptional induction of several NCAM isoforms along with FGFR1 was observed, implicating a mechanistic link between NCAM/FGFR1 signaling and induction of EMT. These assumptions were further supported by the inhibition of the EMT program after specific blocking of FGFR1 signaling responses by PD173074. Moreover, there was evidence for an in vivo TGF-β1 pathway activation with EMT response signal signatures in diseased human kidneys in correlation to impaired renal excretory functions. Conclusion: Modulation of NCAM/FGFR1 signaling blocks the EMT program in cultured human proximal tubular epithelial cells. NCAM/FGFR1 signaling appears to be involved in initial phases of TGF-ß1 initiated EMT of tubular cells and thus could contribute to maladaptive repair and parenchymal damage during renal fibrosis.

Introduction: Primary sinovial sarcoma (PSS) is a very rare malignancy in lung, although lung is the most common organ-based site for metastatic synovial sarcoma. It is a high grade malignant mesenchymal neoplasm showing variable epithelial differentation and characterized by a balanced translocation t(X, 18). Case report: We report a case of a 67- year-old male presented with long lasting cough, chest pain and fever. Chest radiography and computer tomography scan revealed a large well defined lung mass invading hilum of the lung, which was indicated for the lung resection. Histopatological examination showed high-cellulare spindle cell malignant tumor. Immunohistochemically,neoplastic cells were positive for vimentin, TLE-1, Bcl-2, focally positive for cytokeratin AE1/AE3, cytokeratin 7, cytokeratin 18, cytokeratin 19, EMA, calponin and negative for CK 5/6, S100, STAT6, CD34, SMA, Desmin, CD99 and CD 117, which determinate a diagnosis of monophasic PSS of lung. Specific translocation t(X, 18) was detected by fluorescent in situ hybridization (FISH). Conclusion: Diagnosis of PSS in lung may create various diagnostic challenges. It can be established only after exclusion of metastatic sarcoma of other localizations and spindle cell primary lung malignancies. Molecular testing proved to be very helpful or necessary when spindle cell synovial sarcoma was recognized in uncommon sites.

Introduction: Hyperostosis frontalis interna is progressive, irregular thickeness of the frontal bone s internal table. It has been associated with a wide range of syndromes, though we know now that it can occur as an independent entity. Etiology is still unclear, but the most probable hyphothesis is alteration of steroid sex hormons, proven in high frequency among postmenopausal women. It is mostly asympthomatic or it has been accidentally diagnosed after endocranial trauma or postmortal. Because of its progression, in some cases it can cause cerebral cortical compression and varible neurological symptoms. Case report: We present a case of 27 years-old women, hospitalized in Neurosurgical Clinic, because of severe headaches lasting for 3 month. The head CT scan showed a massive, expansive lesion in right frontal bone, with spearing of the superior sagittal sinus and skull midline. Bone bulging does not affect outer table of the frontal bone, but only diploic space and inner table. Pathological analysis of the frontal bone showed marked trabecular thickening both cortical and spongy bone, with corticalisation of spongy bone. Haversian canals showed reduction in number and size, and light fibrosis. The lamellas of the spongy bones were enlarged, with prominent interlamellar bridges and the reduction of interlamellar spaces.In purpose of closer determination of etiopathogenesis,we did immunohystochemical analysis of Estrogen, Progesteron and Androgen receptors in dural tissue. Immunohistohemical stain confirmed nuclear expression of Estrogen and Androgen receptors in dural fibroblasts.Progesterone receptors were negative on immune stain. Conclusion: These results could support the hormonal theory of the cause of hyperostosis frontalis interna.

Aim: Determination of cut-off value for Ki67 index, it’s corelation in luminal breast carcinoma with patient’s age, tumor size, histological grade (HG), and expression of estrogen (ER) and progesterone (PR). Introduction: Breast cancer is a heterogeneous disease characterized by different morphology, immunohistochemical profile, clinical course and response to applied therapy. Ki67 proliferative index is one of the prognostic and predictive factors, whose methodological determination and analysis are still unstandardized. Material and Methods: Retrospectively, we analysed 120 pathohistological reports of patients who were treated in the period 01.01.2009. until 31.12.2011. at the Oncology Institute of Vojvodina, and to whom immunohistochemistry was proven luminal breast cancer (positive ER and PR, negative HER2), without axillary lymph node metastases. Results: The average patient’s age was 57.42±10.17 years; average tumor size 17.98±6.97mm; recurrence was registered in 8 (6.7%) patients with average recurrence time of 49±20.23 months. “Cut off” Ki67 value of prognostic significance for period without recurrence is 20.75%. For correlation testing χ2 and Kendal τ-b tests were used. It’s shown unsignificant relationship between Ki67 and patient’s age (p=0.401 and p=0.293), as well as the strength of expression ER (p=1.00, p=0.957) and PR (p=0.273, p=0.189). Significant correlation is present for Ki67 with size (p=0.035, p=0.20) and tumor’s HG (p=0.041, p=0.20). Conclusion: Breast carcinoma is heterogeneous disease, so it’s difficult to predict its course and outcome using standard histopathological factors and biomarkers. For prognostic-predictive purposes, it’s necessary to include additional biomarkers, where Ki67 stands out above all, whose high value correlates with factors of bad prognosis.

Aim: Determination of frequency of patologic findings and types of pathologic findings in enteric nerve plexus (ENP) in colon biopsies in children with impaired bowel motility, specifically chronic constipation. Introduction: Chronic constipation is relatively common in children, and is most often a functional disorder that is responsive to dietary regime treatment. Rarely, some cases require biopsy and histopathologic analysis of ENP. Materials and Methods: Research consists of 299 colon biopsies taken from children with impaired bowel motility. Biopsies were analysed in Institute of pathology, Medical faculty in Belgrade, in the period of time from the year 2008 to 2018. Data analysis included standard methods of descriptive and analytic statistics. Results: Number of analysed biopsies was 588. Biopsies were taken from 184(61,5%) boys and 115(38,5%) girls. Most common referral diagnosis for biopsy was Hirschspung’s disease (HD) (153/299, 51,2%). Pathologic changes in ENP were found in 46,1% of patients (138/299). Histopathologic analysis confirmed clinical suspicion for HD in 48,4%(74/153) of patients. Most frequent pathologic finding secondary to HD were immature ganglion cells (26/299, 8,7%), ectopic position of ganglia in muscle layer of colonic wall (6/299, 2%), and unclassified dysganglioses (5/299, 1,7%). In six patients, cause of constipation was eosinophilic proctitis and/or mienteric ganglionitis. Acetilcholin esterase as diagnositic metod was applied in 29 patients. Immunohistochemical analises were used in 24 patients. Conclusion: HD and immaturity of ganglion cells are by far most frequently diagnosed causes of constipation in colon biopsies in pediatric patients. Eosinophilic proctitis and/or mienteric ganglionitis are rare causes of constipation in children.

Introduction: GD is the commonest lysosomal storage disease worldwide. The majority of the patients have Type1 GD which is the non-neuronopathic form of disease. There are data of increased risk of cancer in GD patients, such as: multiple myeloma and other haematological malignancies, hepatocellular carcinoma and renal carcinoma. Factors of cancerogenesis in GD are accumulation of bioactive lipids, alternatively activated macrophages, immune dysregulation, genetic modifiers underlying the GD, splenectomy and enzyme replacement therapy. Extra-osseous soft tissue masses are described in GD patients, like localised deposition od Gaucher macrophages (Gaucheroma). To the best of our knowledge, no other case of extra-osseous soft tissue sarcoma in association with GD has been described in literature. To present very rare case of high grade leiomiosarcoma in association with Gaucher disease (GD). Case report: The case concerns 81 years old female with leucopenia and thrombocytopenia since year 2000. In 2014 she was diagnosed with undifferentiated pleomorphic sarcoma with prominent inflammation on her thigh, which was not completely surgically removed. She was diagnosed with leucopenia, thrombocytopenia and splenomegaly in 2014 on control examination. Bone marrow biopsy was performed and histologically and immunohistochemically was diagnosed GD. The diagnosis was confirmed by enzyme activity test. In 2018 revision of pathohistological finding of thigh tumour was performed. High grade leiomiosarcoma was diagnosed. She is alive and refuses any treatment. Conclusion: GD is rarely diagnosed in older age. All soft tissue masses in GD should be carefully examined because of increased risk of cancer in GD patients.

Aim: To investigate co-expression of ATRX and HIF-1α in kidney neoplasm in relation to its origin. Introduction: A heterogenous group of kidney tumors is believed to arise from a variety of specialized cells along the nephron – proximal tubules [Clear cell Renal Cell carcinoma (ccRCC) and papillary RCC (pRCC)] and collecting tubules [chromophobe RCC (chRCC) and oncocytoma]. ATP-dependent helicase (ATRX) is a chromatin remodeling protein involved in gene regulation and aberrant DNA methylation during cancerogenesis. Activation of hypoxia inducible factor (HIF-1α) is an early event in most RCC following inactivation of the VHL tumor suppressor gene. Material and Methods: A total of 46 kidney tumors (n=33 ccRCC, n=1 mRCC, n=4 pRCC, n=5 chRCC and n=3 oncocytomas) was immunohistochemically analyzed for ATRX and HIF-1α expression. Results: Diffuse and focal positivity of ATRX expression was found in 51.5% of ccRCC, while 54.5% had HIF-1α positivity. Co-expression of ATRX/HIF-1α was not related to nuclear grade and stage of ccRCC. Metastatic ccRCC had strong expression of both markers. pRCC type II showed weak ATRX/HIF-1α expression, while pRCC type I was negative for both markers. Interestingly, all analyzed oncocytomas and chromophobe RCC were negative for ATRX/HIF-1α. Conclusion: Our results suggest that signaling pathways have different patterns of activation/suppression of ATRX/HIF-1α in oncocytomas and chRCC compared to other RCC types. Downregulation or loss of ATRX/HIF-1α coexpression in benign tumors should be further investigated in order to determinate mechanisms of ATRX/ HIF-1α signaling transport renal neoplasm with different origin.

Aim: To determine prognosic significance of RAS oncogene mutation detection in patients with metastatic colorectal carcinoma (mCRC). Introduction: CRC is still third most common cancer of both genders and second cause of death from malignancy in Western countries. In recent years, detecting RAS mutation in mCRC tumor tissue has became an imperative in selecting patients for monoclonal antibodies targeted therapy to Epidermal Growth Factor Receptor (EGFR). Nevertheless, there is no consensus regarding prognostic relevance of determining RAS status in patients with mCRC. Material and methods: Study included 116 patients with surgicaly resected CRC at Oncology Insitute of Vojvodina between January and December 2016. KRAS mutation detection was performed in formalin-fixed paraffin embeded tumor tissue samples processed by real-time chain polymerase reaction (real-time PCR) and applyng Cobas® KRAS Mutation Test and Cobas® 4800 System for detection of mutations in codons 12/13 and 61 of KRAS gene. Results: Average age in tested population was 65 years, with a male gender predominance (66.4%). Presence of KRAS mutation was found in 50.9% patients: 44.8% in codons 12/13, 4.3% in codon 61 and 1.7% in both codons 12/13 and 61. RAS mutated colorectal carcinomas (n=59) compared with RAS wildtype colorectal cancers, were significantly associated with male gender, moderately differentiated tumors, lymphovascular invasion and local nodal metastases. Conclusion: Our results show that, beside predictive, KRAS can also have prognostic significance regarding risk assesment for lymphovascular invasion and presence of local and distant metastases.

Aim: The aim of this study was to investigate the expression of Sox1 and Sox2 transcriptional factors in the subgranular zone (SGZ) of the hippocampus in 8 weeks old 5xFAD mice. Introduction: Transgenic 5xFAD mice represent a model of Alzheimer s disease (AD) characterized by an early deposition of amyloid plaques which disrupt the process of adult neurogenesis in the hippocampus. Certain transcriptional factors such as SoxB1 transcriptional factors are involved in the process of adult neurogenesis, but their roles in neurodegenerative disorders are not fully understood. Material and Methods: Transgenic mice of both genders and their respective non-transgenic controls (n=6 per group) were used in the study. Proliferating cells and immature neurons were detected by their immunohistochemical expression of Ki67 and doublecortin, while neuronal stem/precursor cells were identified by the expression of Sox1 and Sox2 proteins. Immunohistochemistry and counting were performed on hippocampal paraffin sections. Results: Immunohistochemical analysis showed that 5xFAD mice in the SGZ of the hippocampus have significantly lower numbers of Sox1 immunoreactive cells, while Sox2 immunoreactive cells were lower only in female 5xFAD mice. Furthermore, we have detected a decrease in the number of newly formed neurons in male transgenic mice, while the number of proliferating cells was unchanged when compared to non-transgenic controls. Conclusion: The results of our study show that early changes in the neurogenesis in 5xFAD model occur despite the preserved proliferative potential in the SGZ. Our results clearly indicate the importance of SoxB1 transcriptional factors in the early phases of AD.

Aim: Evaluation of the usefulness of flow cytometry (FCM) serous effusion analysis in a diagnosis of pediatric Non-Hodgkin lymphomas (NHL). Introduction: Serous effusions are often the first, life-threatening manifestation of pediatric NHL. FCM immunophenotyping of effusions with cytological analysis could help in diagnosis of NHL, and thus enable fast initiating of cytoreductive therapy. Material and Methods: FCM analysis of serous effusions obtained from 17 children and adolescents hospitalized in Mother and Child Healthcare Institute of Serbia under clinical suspicion of NHL using the standardized panel of monoclonal antibodies: CD19, iCD79a, CD20, CD10, iIgM, kappa/lambda, iCD3, sCD3, CD7, CD2, CD5, CD4, CD8, CD1a. Cytological examination was performed on May-Grunwald-Giemsa stained slides. The results were correlated with histopathological findings of available tumor biopsies. Results: Precursor T-cell (T-III/T-IV) phenotype was confirmed in 5 samples. In 7/9 samples with mature B (B-IV) phenotype, FAB L3 cytomorphology indicated Burkitt lymphoma (BL), and in 2/8 suggested diffuse large B-cell lymphoma (DLBCL). Tumor biopsy was available in 7/14 patients and in all cases preliminary diagnosis was confirmed. In 3 patients with no malignant cells in effusions, FCM and cytomorphologicaly only reactive changes were observed, and diagnosis had to be made by tumor biopsy (BL 2 patients, DLBCL 1 patient). Out of 7 patients diagnosed only by FCM and cytological analysis, 6 achieved a remission of the illness. Conclusion: FCM detects NHL cells in malignant serous effusions fast and accurate. In combination with cytological analysis, FCM is sufficient for diagnosis in most cases, allowing rapid initiation of therapy.

Aim: We present a rare case of simultaneous occurrence Concurrent occurrence of acute myeloid leukemia (AML) and monoclonal plasmacitosis (MP): Introduction: The simultaneous occurrence of acute myeloid leukemia (AML) and monoclonal plasmacytosis (MP) in bone marrow (BM) biopsy in patient without previous exposure to chemotherapy is very rare. Case report: We reported the case of a 45-year-old woman who presented with fatigue, fever, maculopapular rush, weight loss and bone pain. She was admitted to the Haematology Department of University CHC Bezanijska Kosa in November 2015. The complete blood count showed: white blood cell count 33x109/l (neutrophils 31%, myeloblasts 12%, monoblasts, promonocytes and monocytes 53%), hemoglobin 77gr/l, platelet count 34x109/l and 5% blastic cells in the peripheral smear. Serum immunoelectrophoresis showed increased monoclonal IgA (IgA 9,98, IgM 2,29, IgG 10,1), kappa/lambda 0,84, Beta2microglobulin 3,16. Biochemistry showed elevated creatinin level 93umol/l, uric acid 412 umol/l, high LDH 876U/l and sedimentation rate (50mm/h). Urine electrophoresis showed monoclonal heavy and light chains lambda type. Liquor immunophenotyping showed neuroleukemia. Skeletal survey showed no lytic lesions. The BM aspiration revealed around 10% plasma cells and BM biopsy showed infiltration by 20% monoclonal plasma cells (kappa-/lambda ) with 30% infiltration by cells with monocytes differentiation (without excess of blasts). Biopsy of skin lesion 17 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. revealed myeloid sarcoma. Cytogenetic analysis detected normal karyiotype with FLT3 and NPM mutation. The patient was diagnosed as AML-M5, administered with induction therapy (DA Cytosar) and consolidation therapy (HiDAC) and she was successfully treated with allogenenic stem cell transplantation. She achieved and maintained complete remission. Conclusion: Monoclonal plasmacytosis of BM should be carefully examined due to possible association with other haematological malignancies

Aim: The aim of the study is to classify tumors into four main molecular subtypes using immunohistochemistry and in situ hybridisation methods, as well as to determine frequency of different carcinoma subtypes. Introduction: Four molecular subtypes of breast carcinoma can be identified: Luminal A and Luminal B (hormone receptors positive), HER2 enriched (HER2 overexpression) and Triple Negative / Basal-like (absence of HER2 amplification and steroid receptors expression). Materials and Methods: Cross-sectional study, conducted in Institute of pathology, Medical Faculty, University in Belgrade, during the ten months period in 2017, included 337 patients with diagnosed breast carcinoma. Using the methods of immunohistochemistry, all four markers (estrogen, progesteron, HER2, and Ki67) were evaluated on breast carcinoma tissues. As additional methods, “in situ” hybridisation (FISH, SISH) was used in cases if HER2 oncoprotein results on immunohistochemistry were equivocal. Results: Luminal A, Luminal B, HER2 enriched, and Triple Negative carcinomas were present in 133 (39,45%), 147 (43,62%), 22 (6,5%), 34 (10,08%) cases, respectively. For 55 (16,35%) cases, in situ hybridisation had been done in order to classify carcinomas in proper molecular subtype. Conclusion: Frequency of molecular subtypes of breast carcinomas in our study are similar to literature data for European countries. Breast carcinoma subtyping has prognostic and predictive values for further patient treatment, and should be done in institutions with adequately trained personnel and equipment, in order to achive best results in the shortest time.

Aim: We present an unusual case of malignant gastrointestinal stromal tumor filling the entire abdominal cavity. Introduction: Gastrointestinal stromal tumors are the most common mesenchymal neoplasms of the gastrointestinal tract. Small tumors are generally benign, but large tumors disseminate to the liver, omentum and peritoneal cavity. They rarely occur in other abdominal organs. Material and Metods: The operative material consisted of segment of ileum where a tumor mass of 8 cm was found originating from the wall and fragments of small intestinal serosa and omentum where multiple nodular tumor mases were found. Representative samples were taken, and were paraffin embedded and stained routinely with Hematoxyllin-Eosin. Additionally immunohistochemical analyses were performed using the antibodies c-kit, CD34, Vimentin, CKAE1/AE3, Ki67 and others. Results: Microscopic analysis revealed a malignant gastrointestinal stromal tumor with a high mitotic rate of 58/50 HPF mitoses. However clinical reports stated that an additional large 12 cm tumor mass was found in the liver that was not removed. Conclusion: Gastrointestinal stromal tumors are the most common gastrointestinal mesenchymal neoplasms but presence of multiple tumor masses on various organs and sites is rare. Presence of multiple tumors in various organs brings about the issues of possibility of multiple primaries or the proper detection of the original tumor mass.

Aim: The aim of our study was to evaluate status of estrogen and progesteron hormone receptors (ER and PR), and HER2 receptors in diagnostic core biopsy specimens, compared to surgical resection specimens of the same patients after NAC regimens all including trastuzumab. Introduction: Neoadjuvant chemotherapy (NAC) is associated with phenotypic alteration in breast carcinoma, especially with the change of molecular phenotype through the modulation of hormone and HER2 receptors expression. Material and Methods: The study included 35 patients with HER2-amplified locally advanced breast carcinoma that were treated with NAC regimens that included trastuzumab, and which had receptors status determined on pre-NAC core biopsy, and on surgical specimen after the completion of the therapy. Results: Pathological complete response (pCR) was observed in 4 cases (11.4%), while partial response to therapy was noted in 31 cases (88.6%). Invasive breast carcinoma of no special type (NST) was the most common histological type in 27 cases (87.1%), while the most common histological grade (HG) was HG3 in 27 cases (87.1%). There were no noted changes in histological type or grade of the carcinoma. The rates of ER and PR receptors positivity on diagnostic core biopsy compared to post-NAC surgical resection specimens were 61.29% to 67.74% and 48.39% to 64,52%, respectively. HER2 receptors status changed from positive to negative in 2 cases (6,45%). Conclusion: Changes in status of the receptors in breast carcinoma after NAC is significant due to implications in tailored therapy approach, and subsequent modification of adjuvant therapy regimens.

Assessment of relationship between density, perimeter and endothelial surface (“endothelial area”, EA) of vascular spaces (VS) using “ImageJ” analysis and morphological characteristics of adenocarcinoma. Introduction: Colorectal adenocarcinoma invasion involves complex reaction of tumor cells and stroma, whereas angiogenesis is essential for metastatic potential. Material and Methods: 70 resected specimens were reviewed. For each adenocarcinoma histological grade, pathological stage, lymphatic, venous and perineural invasion, growth pattern, metastases in lymph nodes and liver were determined. For visualization of vascular endothelial cells immunohistochemical staining CD31 antibody was used. From each tumor nine fields were photographed: three from invasive front, three from VS highest density and three selected randomly. Computer analysis of images was done using program “ImageJ”. For analysis of EA (EA representing surface area of CD31 positive endothelial cells) program recalculated them as percentage of tested fields (% area). VS density was shown as number VS per 1mm2, while perimeter of VS was shown in Results of all parameters were compared with all above described morphological characteristics. Results: There was no significant correlation between density and perimeter of VS and any of histopathological findings. EA from randomly chosen fields (minimum 0,69%, maximum 10,11%, p=0,016) correlates with the presence of venous invasion. There was no significant correlation between EA from the invasive front and areas of the highest density and any of the histopathological findings. Conclusion: Assessment of vascular endothelial surface area is only vascular parameter which positively correlates with prognostic parameters that could indicate worse outcome.

Aim: The goal was to examine the effects of direct exposure of stomach tissue to  acrylamide. Introduction: Acrylamide is a toxic chemical substance discovered in foods rich in starch, prepared at high temperatures. Material and Methods: The research was carried out 6 groups of 5 experimental animals (Wistar rats). Two control groups orally implicated distilled water. Two experimental groups orally administrated PLENARNE USMENE PREZENTACIJE 21 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. acrylamide in a daily dose of 25 mg/kg, and two dose of 50 mg/kg. Three groups were sacrificed after 24h and three after 72h, On histological gastric tissue material is applied qualitative and semi-quantitative histological analysis, stereological measurements of individual compartments of the stomach wall, linear measuring the number of mast cells in the mucosa and submucosa. Results: Slight damage of the surface epithelium, accompanying mild inflammatory reaction and degranulation of mast cells are results of histological damages in the stomach tissue. Reconstruction of the epithelium follows directly toxic effect on epithelium, and it is confirmed by the presence of immature form of mucoproductive cells. Examined inflammatory and degenerative parameters show a positive correlation with respect to dose and/or a time of exposition to acrylamide. Conclusion: Understanding the mechanism of action of acrylamide allows to apply adequate prevention and make an appropriate choice of therapeutic methods.

Aim: The aim of the study is to investigate the effect of prenatal treatment of metronidazole on the process of cerebellar foliation in the guinea pig. Introduction: The use of metronidazole in pregnancy still occupies very different attitudes among doctors who prescribe them. Neurotoxicity is a common side effect in the use of metronidazole, but there is still no clear animal study. Materials and Method: We studied 42 guinea pig fetuses obtain from 12 pregnant guinea pig dams.  Dams were separated in experimental (6) and control6) group(K). Experimental group(E) received from 42-49 day of gestation at every PLENARY ORAL PRESENTATION 22 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. 12 hour (at 8 a.m. and 8 p.m.) subcutaneous injection od saline solution of metronidazole in concentration of 28 mg/kg.  Control group received using same time protocol just saline solution. At 50th day of gestation all dams were sacrified and fetuses were used for further analysis. In each  fetuses (22 in E, and 20 K group). Firstly we make gross picture of the brain and secondary we separated vermis region for further histological and immunohistochemical analysis. Results: An analysis of macroscopic images of the dorsal side of the cerebellum in the experimental group of individuals revealed malformations in 15 out of 22 fetuses (68.2%). The most affected folia was VII and characterized with aberrant diagonal positioning along the entire dorsal side of the vermis. Other folia were partial presence, with micro, and agyria changes. Conclusion: Prenatal use of metronidazole in guinea pigs during the 42-49th day of gestation cause disturbance in cerebellar foliation.

Aim: The aim of our study was to investigate hepatoprotective properties of bile acids (BA), natural ursodeoxycholic (UDK) and semisynthetic 12-monoketocholic acid (MK) by testing the levels of liver enzymes, malonyldehyde (MDA) as an indicator of hepatocyte oxidative damage, activity of antioxidant enzymes, histological analysis of liver tissue and immunohistochemical analysis of hepatocyte proliferation, activation of nuclear farnesoid-X receptor (FXR), apoptosis and proliferation markers. Introduction: The hepatocyte metabolism is extremely intense, and the processes that take place in hepatocytes are diverse, which makes them highly susceptible to oxidative damage that is the basis of many diseases. Two popular models of oxidative stress induction are the use of ethinylestradiol and the cholestasis model, as well as the use of aloxan and aloxan-diabetes model. Results: Male Wistar rats were used in this study. Control group (K) received only saline (0.5 ml/kg). Three groups received ethinylestradiol (EE group, 0.5 mg/kg) as monotherapy or in combination with 12-monoketocholic (E-MK, 4 mg/kg) or ursodeoxycholic acid (EPLENARY ORAL PRESENTATION 24 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. UDK, 25 mg/kg). In order to increase the level of oxidative stress, in three groups monodose of aloxan was administered in addition to ethinyl estradiol, either alone (AE group), or treated additionally with 12-monoketocholic (AE-MK) and ursodeoxocholic (AE-UDK) acid at the above dosages. Liver tissue was sampled for histological analysis and biochemical analysis of liver enzymes and bilirubin (AST, ALT, GGT, bilirubin total (BLRu) and direct (BLRd)), oxidative damage (MDA), levels of antioxidant enzymes catalase (CAT), glutathione-peroxidase (GSH-Px), glutathione-reductase (GSH-R), glutathione-S-transferase (GSH-ST). In groups EE and AE, AST, ALT and GGT levels were significantly elevated compared to K group, while the use of bile acids led to a significant reduction in activity of all enzymes as well as total and direct bilirubin. Both acids have the strongest hepatoprotective effect in terms of AST activity and lowering the total bilirubin concentration, whose values using both UDC and MK were lowered to the level of K-group. In terms of ALT and GGT, bile acids showed slight selectivity. Histological analysis of liver in groups EE and AE showed hepatocytes with karyopycnosis and cytoplasmic acidophilia, which indicate the apoptotic cell death, as well as changes in individual hepatocytes corresponding to feathery degeneration. All of these changes weren`t observed in the liver of animals treated with UDK and MK. By analyzing Ki-67 in the tissue of the control group, it was found that up to 5% of the cells exhibited nuclear expression, and a value of 10% was taken as the limit for the enhanced proliferative activity of hepatocytes. In groups that received only hepatotoxic substances, ethinyl estradiol and aloxan, the expression of ki-67 was low as in controls. Animals whose hepatocyte damage was caused only by ethinylestradiol, reflected hepatoprotective properties of bile acids by enhancing hepatocyte proliferation (E-UDK in 50%, and E-MK in 100% of animal). Similar hepatoprotective effects of tested bile acids was seen when comparing group treated with both hepatotoxic substances (AE), and groups cotreated with UDK (all animals had an increased proliferative activity) and MK (80% of animals). Accumulation of hydrophobic acids in ethinylestadiol-induced cholestasis enhances oxidative damage and induces apoptosis, but also activates the expression of nuclear farnesoid-X-receptor. In the K-group, 50% of the animals showed low expression of FXR, and moderate and high expression was present in 25% of animals. In EE and AE groups, high FXR expression was significantly increased (75%, ie 60%) indicating the accumulation of hepatotoxic acids that raise the level of oxidative damage and induce apoptosis. The hepatoprotective effect of UDK and MK, has been demonstrated by the reduced content of hydrophobic acids, ie, the reduced expression of FXR. In groups in which hepatotoxicity and oxidative stress were induced only by ethinylstradiol and were treated with UDK and MK, not a single animal had high FXR expression, and a number with weak expression increased significantly. Similarly, in groups that received two toxic substances and analyzed acids (AE-MK, AE-UDK) 60% of animals had weak FXR expression, which is statistically not different from the control group. Groups treated with ethinylestadiol and aloxan had a significantly higher concentration of malonylaldehyde (MDA), an indicator of oxidative hepatocyte damage. The use of UDK and MK significantly reduced the concentration of MDA, but not to the control group level. The activity of antioxidant enzymes in the EE group did not differ statistically from control, and the use of UDK and MK resulted in significant changes. In the group AE where oxidative stress is enhanced, the activity of all analyzed enzymes is significantly decreased, and the application of UDK and MK significantly enhances the activity of the antioxidative enzymes. In order to examine the hepatotoxicity of oxidative stress and hepatoprotective properties of UDK and MK, the expression of the p53 and Bcl-2 family of proteins was studied. Hepatocytes of the control group did not express the proapoptotic Bax, while in the EE and AE groups, weak expression was present in 50% and 40%, respectively. Using UDK in both EE-UDK and AE-UDK models, Bax was completely reduced to the level of control group, ie no Bax expression was observed. The use of MK in both models led to suppression of Bax (EE-MK, 25%; AE-MK, 0%). The antiapoptotic Bcl-2 protein was detected in the control group in 25% of the individuals, unlike the EE and AE groups, where the expression was completely suppressed. By promoting the expression of Bcl-2 (E-UDK in 25% of individuals, AE-UDK in 40% of individuals), the UDK showed a stronger hepatoprotective activity compared to MK which in the AE-MK group only led to an increase in expression by 20%. In the control group p53, expression was recorded only in the cytoplasm, in 50% of animals, while in the EE and AE groups p53 had nuclear expression in 25%, ie 20% of the individuals, which speaks in favor of more intensive apoptotic action and induction of transcription of other proapoptotic proteins. In groups where UDK was administered, p53 was not present in either the nucleus or the cytoplasm, while MK reduced the expression to the control group level. Conclusion: Ursodeoxycholic and 12-monoketocholic acid have great antioxidative and antiapoptotic capacity, which explains their hepatoprotective effect.

The fourth Edition of the World Health Organization’s Classification of Tumours of Endocrine Organs from 2017 introduced some novelties based primarily on genetic research, compared to the previous edition from 2004. The most common primary tumors of the adrenal cortex are adenomas, and of the adrenal medulla are pheochromocytomas. As such, these tumors have their specificities in pathohistological diagnosis, by which they differentiate from tumors of other localizations. In the daily work of the pathologist dealing with the diagnosis of the tumor of the adrenal gland, differential diagnostic problems can occur, where biological behavior of the tumor cannot be predicted based on its morphological appearance; this primarily refers to cortical neoplasms and phaeochromocytomas. Scoring systems are used in these cases, Weiss and Lin-Weiss-Bisceglia criteria for cortical neoplasms, and PASS criteria for pheochromocytomas, where by recognizing the suggested morphological parameters, value or score is obtained which predicts the biological behavior of the tumor, i.e. differentiates benign from malignant neoplasms. The accepted algorithm of diagnostics of adrenocortical neoplasms also includes immunohistochemical staining for ki-67, which potentially can differentiate adrenocortical adenoma from carcinoma. Despite the introduction of these scoring systems, the only safe indicator of malignancy is still distant metastasis.

Pancreatic cancer is high aggressive malignant neoplasm with very poor prognosis and about only 5% a five-year survival. It is the fourth leading cause of cancer death in USA, as well in Serbia, although in terms of the incidence of this disease the mortality is rising and it takes seventh place. Speaking about the southern Backa, the mortality rate is slightly lower and it is the fifth place, according to the available literature. This can be explained by the lack of highly specific and sensitive diagnostic tests, which makes pancreatic cancer most often detected by an advanced, inoperable stage of the disease (>60%), although surgical resection is the only curative therapy. Analogous to other carcinomas, there is a gradual progression of the pancreatic duct epithelial cells, so every invasive carcinoma arises from the previous intraepithelial neoplasia. There are three different types of common precancerous lesions known for pancreatic cancer which clinical detection and treatment can stop the progression to invasive cancer and reduce mortality. The first one is Pancreatic Intraepithelial Neoplasia (PanIN), and the other two types of precancerous lesions are both larger fluid-filled types- Intraductal Papillary Mucinous Neoplasms (IPMNs) and Mucinous Cystic Neoplasms (MCNs). PanIN is an asymptomatic, small (usually <5 mm in diameter), nonfluid flat or papillary lesion arising in the small intralobular pancreatic ducts. It is the most common and important precursor of invasive pancreatic carcinoma. Histologically, it’s consisted of columnar to cuboidal cells with varying amounts of mucin. Accordingly to different degrees of cytological and architectural atypia, PanIN is classified into three grades- low, intermediate and high grade. PanIN-1A (flat) and PanIN-1B (papillary) are low grade lesions with minimal atypia, while PanIN-2 belongs to intermediate lesion showing mild to moderate atypia with frequent papillae. PanIN-3, also referred as “carcinoma in situ”, is characterized by severe cytological and architectural atypia amd it can be flat, papillary or cribriform pattern. The immunohistochemical profile of PanINs vary with the grade of dysplasia. Low grade lesions show positivity for gastric foveolar mucin MUC5AC, pyloric gland mucin MUC6, while MUC1 is almost exclusively expressed by high grade PanINs lesions. Among fluid-filled types of precancerous pancreatic lesions, IPMNs are the most common lesions. IPMNs vary in their location and size within the pancreas size, and these two features correlate with how dangerous they are. Because they can be detected by imaging procedures, it is possible to detect them before they become cancer. They are most often detected in patients who are routinely monitored due to a high familial risk, or incidentally in people who were imaged for another reason. Those lesions belong to group of the heterogeneous group of cystic pancreatic lesions because papillary epithelial proliferation and mucin production lead to cystic dilatation of involved ducts. SPECIJALNA SESIJA: KATEDRA ZA PATOLOGIJU MEDICINSKOG FAKULTETA, UNIVERZITETA NOVI SAD, SRBIJA 29 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. Those lesions are slightly more common in the head and uncinate process (55%) compared with the body and the tail of the pancreas and about 30% of them are multifocal. IPMNs are subdivided into main duct IPMN (MD-IPMN) which are localized in the main pancreatic duct and measured ≥5 mm, while the other branch duct type (BD-IPMN) is >5 mm in diameter andcommunicates with the main pancreatic duct but it is uninvolved by the process. Mixed IPMN combines both types. Microscopically, the lining epithelial component is represented by tall mucin producing columnar cells but lack the “ovarian-type” seen in mucinous cystic neoplasms. Like PanINs, IPMNs are graded on the basis of the greatest degree of dysplasia into low grade, moderate and high grade dysplasia or carcinoma in situ and IPMN with associated invasive carcinoma. Many studies have showed that approximately one-third of patients with IPMN are associated with invasive carcinoma, so precise basement membrane micro analysis is an imperative. Accordingly to their histological characteristics there are intestinal, pancreatobiliary, oncocytic and gastric subtype of IPMN with different immunohistochemical profiles. Intestinal-type IPMN is characterized by tall columnar cells with elongated nuclei and amphophilic cytoplasm and MUC2, MUC5AC, MUC4 and CDX-2 positivity. In contrast, pancreatobiliary subtype of IPMN is characterised by branched papillae with high grade intraepithelial neoplasms and MUC1 And MUC5AC immuno positivity. The third are oncocytic type, predominantly occurs in main duct and presented with a complex branched papillary structures covered by oncocytic cells mixed with goblet cells and mucin-containing cells. This type shows diffuse positivity for MUC5AC, MUC6 and focal positivity for MUC1 or MUC2. The gastric subtype is low grade lesions mainly found in branched ducts and characterised by papillae covered by foveolar glandular epithelium with MUC5AC and sometimes MUC6 positivity. The last and the most infrequent type of pre-cancerous pancreatic lesions are MCNs. The principle difference between IPMNs and MCNs is how they look under a microscope, and how they behave in the patient. These cystic lesions are almost solitary, typically located in the pancreatic body and tail. Grossly, this lesion can grow very large, it is usually septated and with fibrous pseudocapsule often with calcifications. Histologically, the epithelium og noninvasive MCN is consisted of columnar cells with varying degree of dysplasia (low grade, moderate and high grade dysplasia) and underlying ovarian-like stroma. Immunohistochemically, thode epithelial cells are EMA, CEA, MUC5AC, MUC2, cytokeratins 7, 8/18 and 19 positive, while the underlying ovarian-like stroma shows ER, PR, vimentin and SMA immune reactivity. Precancerous lesions of the pancreas are important changes whose visualization and detection in a significant number would reduce the incidence of pancreatic cancer and, consequently, the mortality of this highly aggressive neoplasm with an unfavorable therapy outcome.

Pathologists often have a dilemma is a lymph node biopsy reactive or corresponds to a lymphoproliferative or other malignant disease. In everyday routine work, we rely on morphologic criteria and immunohistochemical analyzes. In better-equipped labs additional cytogenetic and molecular methods are used if morphology and immunohistochemical analyzes are not sufficient for getting correct diagnoses. It is important to know clinical presentation and the opinion of a clinician who runs the case. In reactive lymph nodes general morphology is mostly preserved. Distribution of B and T cells, histiocytes, dendritic cells and proliferation is adequate. Foreign cells are not present. Ways of reaction in lymph nodes are follicular hyperplasia, paracortical expansion, sinus histiocytosis and granuloma formation. If metastases are present, most often from carcinomas and melanomas, the initial deposits are usually sub capsular or less often in sinuses. One should be careful to differentiate sinus histiocytes and metastatic tumor cells, what can easily be verified by immunohistochemical stains.If it is a lymphoma, one should decide is it a Hodgkin or a non-Hodgkin lymphoma. In non-Hodgkin lymphomas, one should decide between small cell and large cell lymphomas. In non-Hodgkin lymphomas, tumor cells are dominant and background inflammation is scant and mostly consisted of small T cells and rare histiocytes. In T cell lymphomas background inflammation can be quite various. In Hodgkin lymphomas background inflammation most often is various and almost always outnumbers tumor cells. Tumor cells are large, with lobulated or multiple nuclei and conspicuous nucleoli. The immunophenotype is usually clearly different from non-Hodgkin lymphomas. The differentiation of small cell and large cell non-Hodgkin lymphomas is easily made by comparing cell sizes. If tumor cell size is closer to size of histiocytes or endothelium it is a large cell lymphoma, but if it SPECIJALNA SESIJA: KATEDRA ZA PATOLOGIJU MEDICINSKOG FAKULTETA, UNIVERZITETA NOVI SAD, SRBIJA 31 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. is closer to small lymphocytes and red blood cells it is a small cell lymphoma. Differentiation of small cell lymphomas is based on morphology, distribution of cells and on immunophenotype. Differentiation of large cell non-Hodgkin lymphomas requires immunohistochemical analyzes because morphology is often very similar among entities. Correct diagnosis is important due to application of optimal therapy and reaching the best prognosis for the patient.

In preparation for the 8th edition of the TNM classification for lung cancer the International Association for the Study of Lung Cancer (IASLC) collected data on 94,708 cases of lung cancer diagnosed between 1999 and 2010, donated by 35 institutions in 16 countries. After exclusions, 77,156 remained for analysis: 70, 967 cases of non-small cell lung cancer (NSCLC) and 6,189 cases of small-cell lung cancer (SCLC). Analysis of the cases of NSCLC has allowed proposals for revisions to the T, N and M descriptors and TNM Stage groupings. Size remained an important determinant and a descriptor for all of the T categories. A new cut points at 1 and 4 cm have been proposed and as a result new T categories have been created: T1a ≤1 cm, T1b > 1 to 2 cm, T1c > 2 to 3 cm, T2a > 3 to 4 cm, T2b > 4 to 5 cm, T3 > 5 to 7 cm and T4 > 7 cm. However, measuring precise tumor size can be challenging since it is known that tumor gross size depends on whether the size measurement is performed on fresh or formalin-fixed specimen. In about 10% of cases, formalin fixation can cause down-staging of pathologic T category as a result of tumor shrinking. Tumors invading the diaphragm have been reclassified as T4, and tumors extending within 2cms of the carina without its invasion, or tumors associated with collapse or consolidation of the whole lung have been down-staged to T2. Tis and T1mi were introduced for adenocarcinoma in situ, squamous cell carcinoma in situ and minimally invasive adenocarcinoma, respectively. Visceral pleural invasion, defined as the involvement of its elastic layer, remains unchanged as T2 category, but specific analysis of visceral pleural invasion, showed that there is two types of invasion: PL1 where tumor invades beyond the elastic layer and PL2 where tumor invades pleural surface and that these two had different prognosis, PL2 being associated with the worst outcome. Elastic stains are recommended to clarify the status of visceral pleural invasion for cases in which initial hematoxylin-and-eosin-stained slides failed to show presence of invasion. Mediastinal pleura invasion disappears as a T descriptor. N categories remained the same as in 7th edition. 8th did not bring guidelines about the minimum number of lymph nodes that should be assessed for pathohistological analysis. In M descriptor category M1a retained, while M1b has been reassigned to describe a form of limited disease with a single metastatic deposit in one distant organ. A new category of M1c has been proposed and it is reserved for situations in which there are multiple metastases in one or more distant sites. Assessment of multifocal lung tumors and the distinction of synchronous primary tumors from intrapulmonary metastases represent an important problem as this decision significantly influences tumor staging, as well as treatment approach. Four different clinical presentation of lung cancer with multifocal lung involvement are described: second primary cancer, intrapulmonary metastasis, multifocal lung adenocarcinoma with ground glass/lepidic features, and pneumonic-type lung adenocarcinoma. The tumors are considered second primary tumor if it have clearly a different histology or have a different radiographic appearance, metabolic uptake growth pattern or different biomarkers. Each tumor is staged separately based on current TNM staging system. The nodules are considered to be intrapulmonary metastasis if exact matching breakpoints are identified by genetic hybridization or have similar clinical features such as radiographic appearance, growth pattern or significant nodal and systemic SPECIAL SESSION: DEPARTMENT OF PATHOLOGY, MEDICAL FACULTY, UNIVERSITY NOVI SAD, SERBIA 32 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. metastasis. TNM staging depends on location of the nodule relative to the primary tumor site. If it is in the same lobe, the tumor is designated as T3, if it is in the same lung, but in different lobe as T4, and it it is in the contralateral lung as M1a. Tumors are considered multifocal lung adenocarcinoma if there are multiple subsolid nodules with at least one suspected or proven to be a cancer. Ground glass nodule <5 mm or lesion suspected to be AAH is excluded. T stage is based on highest T lesion with indicating the multiplicity. Tumor is categorized as a pneumonic-type adenocarcinoma if there is a diffuse pneumonic infiltrate or consolidation with regional distribution. Stage IA is divided into IA1, IA2 and IA3 to accommodate T1a, T1b and T1cN0M0 tumors. All N1 disease is staged IIB except for T3-T4N1M0 tumors which are stage IIIA. A new stage IIIC is created for T3-T4N3M0 tumors and stage IV is divided into IVA (M1a and M1b) and IVB (M1c). In conclusion, multi-disciplinary approach and the close cooperation among medical and radiation oncologists, pulmonologists, surgeons, radiologists and pathologists is important in properly staging of lung cancer as well as, in treatment plans.

Lung cancers are the most common cause of morbidity and mortality from malignant tumors in the World. The neodjuvant therapy in patients with locally advanced (IIIA-IIIB) lung cancer and affected N2 lymph nodes is one of the modes of multimodal treatment of patients with non-small cell lung cancer (NSCLC) in order to improve the outcome of their treatment. This involves converting patients from a higher to a lower stage of the disease - “downstaging”. There has been no significant connection between some forms of tumor response and types of therapy. Given the importance of complete pathological responses and tumor regression in the prediction of treatment outcomes, finding this relationship is of importance for the design of future neoadjuvant trails. In determining the histological tumor regression is very important measurement of area of residual tumor (ART). As the size of the tumor is one of the prognostic factors in patients with NSCLC who did not receive neoadjuvant therapy so the measurement of ART, as opposed to the macroscopic size of the tumor, one of the prognostic factors in patients with NSCLC, who had received neoadjuvant therapy. The ultimate goal of neoadjuvant therapy should be resectability and “downstaging” that could provide overall oncology benefit in specific clinical situations. The main objectives of this research were: to objectively estimate the size of ART in tumor tissue of lung and lymph nodes; to estimate the relation between the surface of ART with the size of the tumor on postoperative surgical material after neoadjuvant therapy; to analyze and estimate the relation between histomorphological parameters in tumor regression induced by neoadjuvant therapy and spontaneous tumor regression in tumors of the lung and lymph nodes in the postoperative surgical material and depending on the histological type of cancer; to estimate the relation between clinical response to neoadjuvant therapy according to criteria of the World Health Organization and histological parameters in lung tumors and lymph nodes in the postoperative surgical material after neoadjuvant therapy; to estimate the correlation of the pathological ypTN with clinical ycTN stage of the disease and the degree of tumor regression induced by neoadjuvant therapy and pathological ypTN and estimation of the relation between clinical and pathological involvement of N2 lymph nodes after neoadjuvant therapy. Measurement of the total size of the preserved ART is the most important objective parameter in the assessment of the grade of tumor regression. Size of residual tumor did not correlate with the size of the tumor after neoadjuvant therapy. There was a significant difference in the histological picture of tumor regression induced by neoadjuvant therapy and spontaneous tumor regression. There was no significant difference between the histologic type of tumor and histological tumor regression. There is no significant correlation between clinical response and the grade of tumor regression after neoadjuvant therapy. There is no correlation between clinical and pathological staging of the diseaSPECIAL SESSION: DEPARTMENT OF PATHOLOGY, MEDICAL FACULTY, UNIVERSITY NOVI SAD, SERBIA 34 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. se after neoadjuvant therapy. There is no correlation between the grade of tumor regression induced by neoadjuvant therapy and ypTN stage of the disease. There is no correlation between the clinical and the pathological involvement of the N2 lymph nodes to neoadjuvant therapy. The grade of tumor regression and measurement ART after neoadjuvant therapy determined by histopathological analysis of the resected tumor is the most objective criterion for evaluation of chemotherapeutic response and prediction of treatment outcome in patients.

Lung carcinoma is the leading cause of increases in the morbidity and mortality rates of malignant diseases worldwide. Adenocarcinoma has been the most common histological type in the last decades due to: changes in the tobacco industry, smoking habits and the use of immunohistochemistry. Among more than half of patients, lung adenocarcinoma is diagnosed in an advanced stage of the disease. The discovery of mutations in epidermal growth factor receptor (EGFR) in lung adenocarcinoma is a major advancement in molecular pathology and a new approach to the treatment of these patients. Patients with EGFR mutated lung adenocarcinoma receive a targeted therapy (Tyrosine Kinase Inhibitors-TKI) which leads to improvements in disease prognosis and quality of life. Real-time polymerase chain reaction (PCR) is the most widely used and most reliable method since it requires a minimum amount of starting material and allows the amplification of the desired DNA segment up to a billion times. In this way, deletions in exon 19 are detected in approximately 90% of cases, more often in women, non-smokers and in the territory of Asia. The following may be used for EGFR testing: fresh tissue, fast-frozen tissue, tissue molded into paraffin blocks after fixation in formalin and cytological material obtained by scraping from glass tiles. Tissue processed by decalcination, acid treated or heavy metal treated tissue should be avoided. Although surgical samples represent the golden standard in determining EGFR mutations, the results obtained are compatible with the results obtained by bronchoscopic biopsy and thus eliminate the need for invasive diagnostic procedures. Bronchoscopy is an invasive diagnostic method, whose objectives are to diagnose lung tumors, determine the endoscopic spread of the disease and assess tumor operability. The presence of a tumor may be indicated by a different bronchoscopic aspect of the endobronial mucosa. The sensitivity and specificity of this method depends on: bronchologist’s skills, endoscopic findings, the number of biopsy samples, the professional competence of pathologist-cytologist and the obtained tumor amount. The tumor amount is generally small and depends on the histological type, endoscopic findings, sampling technique and the presence of other cells. It is recommended to take three to five biopsy samples, used for diagnosing but also for molecular testing. Targeted therapy is applied based on the obtained results. Given that biopsy samples molded in paraffin are cut into multiple histological sections, and that the tumor amount decreases, it is necessary to minimize the “consumption”. The concentration of isolated DNA does not differ among patients with wt EGFR and mutated EGFR adenocarcinoma. To date, there has been no consensus regarding the number of tumor cells necessary to determine EGFR mutations, and it is recommended to take samples with a minimum of 200 to 400 tumor cells. Invalid results obtained by using the PCR method are most commonly the result of a small number of preserved tumor cells in a biopsy sample. Blood and necrosis may be limiting factors for molecular testing, but not exclusion factors for the same. Bronchoscopic biopsy sample is adequate for the determination of EGFR mutations because the majority of biopsy samples have more than 100 tumor cells, the difference between the concentration of isolated DNA in EGFR mutated and wt EGFR adenocarcinomas is not statistically significant, EGFR mutations are also detected in samples with a small number of tumor cells when using highly sensitive tests.

Aim: The aim of this study was to investigate the association of local tumor survivin expression and serum concentration with clinical and histopathological parameters in melanoma patients. Introduction: Survivin is a multifunctional protein abundantly expressed in tumors of various types, including melanoma. There are still sparse data regarding relationship of melanoma cell survivin expression with accepted histopathological characteristics as well as serum concentration. Material and Methods: The level of survivin expression was determined immunohistochemically in tumor tissue and with ELISA test in the serum of 84 melanoma patients with melanoma. Results: Survivin expression was significantly higher in the patients whose tumor had ulceration, higher mitotic index, higher Clark and Breslow stage, that made vascular invasion or spread through lymphatic vessels in primary tumor, and in the patients with metastatic disease. The patients with high survivin expression score had almost double shorter disease free interval DFI comparing to those with weak local survivin expression and a small number of survivin cells (9 - 7 vs 19 - 13 months, respectively). The degree of tumor infiltrating lymphocytes presence in tumor tissue was significantly inversely associated with serum survivin concentration. Conclusion: Conclusion Survivin expression in tumor tissue and its serum concetration significantly correlate with clinical and histopathological parameters. Serum levels could be important in initial follow-up as indicators of those patients that would have aggressive local tumor growth and spreading.

Aim: To highlight the widespread metastatic potential of the cutaneous melanoma, as well as its tendency for unusual presentation of metastatic disease. Introduction: Melanoma is an aggressive, highly malignant disease that is derived from melanocytes. The incidence of melanoma is significantly increasing. Melanoma has a strong tendency for metastasis. After primary excision of tumour, about 30% of all patients shall develop distant metastasis within first 5 years after tumour diagnosis. Case report: A 48-year-old female patient had undergone a hysterectomy because of myomatous uterus. After pathohistological examination metastasis of melanoma was diagnosed in one of multiple leimyoma. Diagnosis was confirmed with positive immunohistochemical staining with MART1 and S100 protein. Insight into the medical records, revealed that patient was diagnosed with superficially spreading melanoma (Clark IV, Breslow III) on skin above her left breast, as well as 2 regional tumour-involved lymph nodes (pT3aN2bM0), 2 years prior to this hysterectomy. Uterine leiomyoma was the first diagnosed distant metastasis of cutaneous melanoma. Diagnosis of stadium IV melanoma was established. Conclusion: Melanoma is a particularly aggressive disease with unpredictable evolution, so the occurrence of metastases in unusual and unexpected localizations, as is the distant benign tumour in the presented case, shall probably happen more often in the future.

Aim: Report of two cases of benign skin tumors of different histogenesis with anaplastic lymphoma kinase (ALK) gene rearrangement. Introduction: ALK gene mutation or rearrangement positive tumors (ALKomas) are heterogeneous group in which such genetic finding has diagnostic or predictive value. ALK gene fusions are associated with tumorigenesis of some cutaneous tumors, e.g. plexiform Spitz nevus (PSN) and epithelioid fibrous histiocytoma (EFH). Cases reports: Histological, immunohistochemical (IHC) and genetic characteristics were analyzed in two skin tumors with ALK gene rearrangement proved by fluorescent in situ hybridization (FISH). First case is PSN localized on face in a 8-year-old boy. Tumor was consisted of spindle cells arranged in plexiform growth pattern throughout epidermis and dermis, with following IHC characteristics: HMB45  focally, p16 , Ki67  in 1% of cells. Deletion of CDKN2A gene was not detected in significant number, while ALK gene rearrangement was positive in 37/50 (74,2%) of cells. Second case is EFH localized on shoulder in a 16-year-old girl. This dermal based tumor was consisted of tightly packed large epithelioid cells, which were CD68 , p16 , S100-, HMB45-, SOX10-, CD1a-, Ki67 (<5% of cells). ALK gene rearrangement was positive in 24/100 (24%) of cells, and focal chromosome 2 polysomy was noted. Neither mitoses nor necrosis were present in any of presented cases. Conclusion: Skin ALKomas are tumors with heterogeneous histological and immunohistochemical characteristics, and also with variable extent of ALK rearrangement.

Aim: Our aim is to examine the expression of CD34 in the endometrioid carcinoma (EC) of the uterine body and association of neoangiogenesis with classical clinical and prognostic parameters. Introduction: The incidence rate increases with age, so in about 75% of cases it occurs in postmenopausal women.Material and Methods: On the biopsy samples obtained after the hysterectomy of 36 patients operated from EC of the uterine body, operated in the General hospital Pirot, where applied routine H E and the imunohistochemical ABC method with anti-CD34 antibodies. Based on the expression of CD 34, the microvascular density per mm2 of the tissue was calculated stereometrically. For statistical analysis SPSS software package were used (version 19.0). Results: The significantly pronounced expression of CD34 is present in all cases of tumors in pT2a to pT3a, and in all cases of the IIA-IIIA FIGO stage. A significantly high neoangiogenesis index is present in 72% of moderately differentiated tumors, in about 70% of tumors with moderate/expressed nuclear atypies, in 92% of cases with microvascular invasion and in 76,5% of tumors affecting over 50% of myomterium. CD34 expression is in positive, moderate and significant correlation with invasion of lymph vessels, pathological stage of the tumor and invasion of blood vessels and myometrium (kk=0,636 - 0,587). The correlation between this marker and the histological and nuclear tumor grade is weaker (kk=0,444 and 0,410). Conclusion: Significant association of CD34 with the aggressive phenotype of the EC of uterine body has significant prognostic implications.

Aim: Lobular endocervical glandular hyperplasia is very rare, differential diagnostic difficult entity, which often mimics more aggressive endocervical lesions, with worse prognosis. Introduction: Lobular glandular endocervical hyperplasia(LEGH) is benign lesion with lobular proliferation of endocervical glands and endocervical epithelium without or with minimal nuclear atypia. Lobular structure can be deep into the cervical tissue without stromal invasion, which is important difference between adenocarcinoma of minimal deviation(MDA), well-differentiated endocervical type of adenocarcinoma, similar to LEGH. Case report: The patient is 50 years, with demarcated, nodular, multicystic tumor, localized deep in cervical tissue. Microscopic analysis shows numerous cystically dilated endocervical glands with lobular presentation, surrounded by hyperplastic fibrous stroma. Glandular acini are diffusely localized, with centrally dilated acinus and peripheral smaller tubular structures. Glands are without the presence of architectural changes and cytonuclear atypia, with uniform, endocervical epithelium.Immunochemical analysis showed:CEA- /CA125focal /p16 focal /Er and Pr focal epithelium, diffusely in stroma. Because structures were described deep in the cervical tissue, MDA is considered diagnostically differential. However, glands do not show signs of distortion(architectural atypia), which is important feature of this entity, as well as no pronounced surrounding desmoplastic stromal reaction. Macroscopic tumor is well demarcated, and the CEA marker is negative, which also suggested LEGH. Conclusion: LEGH and MDA show a quiet clinical and histological similarity. Association of LEGH(50%) with MDA and 40% with cervical adenocarcinoma has been demonstrated, suggesting the possible precancerous potential of LEGH. Research of the lesion microenvironment could be an interesting subject for further research.

Aim: Evaluation of pathological estimation results of therapeutic response of primary breast carcinoma to applied neoadjuvant therapy. Introduction: Breast carcinoma is heterogeneous disease, that could be classified into several molecular subtypes by using immunohistochemical analysis and in situ hybridization. Neoadjuvant therapy is applied in cases of locally advanced breast carcinoma and for tumor chemosensitivity evaluation, which is very significant for disease prognosis. Material and Methods: The research was conducted on 35 female patients, that were treated with radical mastectomy in 2017, after IV cycle of neoadjuvant therapy, in Clinical Centre of Montenegro. Before treatment, all patients underwent core biopsy (confirmed invasive breast carcinoma, molecular subtype determined).Pathohistological response of primary tumor to applied therapy was estimated as complete response (pCR), partial response (pPR) and no response (pNR). Results: Average age in examined group of patients was 59,25 years.In 68,58% of cases it was invasive ductal, in 17,14% invasive lobular and in 14,28% mixed invasive carcinoma. Representation of molecular subtypes was (without change in respect to the core biopsy): 34,28% Luminal A, 42,85% Luminal B Her2 negative, 5,71% Luminal B Her2 positive, 8,58% Her2 positive and 8,58% triple negative. In 6 patients (17,15%) pCR was obtained, in 21 (71,42%) pPR, and in 4 (11,43%) no response. In group of patients confirmed for pCR, it was Her2 positive or triple negative carcinoma. Conclusion: Applying of neoadjuvant therapy leads to tumor response to applied therapy. Most commonly it is partial response, while complete response most commonly occurs in Her2 positive or triple negative carcinomas.

Aim: To investigate concordance rate between the results of expression of steroid receptors, human epidermal growth factor 2 receptors (Her2) and determined molecular subtypes in surgical specimens (SS) and samples obtained by core biopsy (CB). Introduction: CB is widely accepted method in the initial diagnosis of breast cancer, but its reliability in determining the status of steroid and Her2 receptors, Ki67 index and molecular subtypes is still a matter of debate. Material and Methods: We analyzed 54 cases of invasive breast cancer, in which the expression of estrogen (ER), progesterone (PR) and Her2 receptors and Ki67 index were determined both in CB and SS. Concordance rate for ER, PR and Her2 receptors expression and molecular subtypes, between CB and SS, was calculated using k-test (p<0.001). Results: The average age of patients was 62. In SS, Luminal A subtype was most commonly diagnosed (48%), followed by: Luminal B Her2-(31%) and TNBC (13%), while Luminal B Her2 and Her2-enriched subtypes were represented by 4% each. Frequencies of molecular subtypes in CB were: Luminal A (41%), LuminalB Her2- (33%), TNBC (15%), Luminal B Her2 (7%), Her2-enriched (4%). Concordance rate for ER receptors was 93.8%(Kappa=0.936), for PR 77.5%(Kappa=0.773), for Her2 80.0%(Kappa=0.78) and for molecular subtypes 80.9%(Kappa=0.753). Conclusion: Statistical analysis showed very good agreement in terms of determined molecular subtypes and ER receptors expression and good agreement for the expression of PR and Her2 receptors. CB represents reliable method for determining the status of expression of steroid and Her2 receptors, as well as tumor molecular subtypes.

Aim: To compare sensitivity, specificity and accuracy of bronchial brushing (BB) and transbronchial needle aspiration (TBNA) cytology in diagnosing lung cancer and to correlate and compare them with corresponding histopathology. Introduction: Lung cancer is a leading cause of death worldwide in both genders. Various cytological techniques such as BB and aspirate, TBNA, bronchoalveolar lavage, transthoracic and pleural puncture can aid in early diagnosis of lung malignancies. Material and Methods: One-year retrospective study included 359 patients with suspected lung cancer who underwent bronchoscopy. During bronchoscopy, cytopathological samples were obtained for smears using BB and TBNA as well as biopsy for histopathological examination that was considered the  gold standard .  All the samples were microscopically examined and statistically analyzed using descriptive methods and non-parametric Kendal-tau correlation coefficient (the level of significance p<0.05). Results: Sensitivity of BB and TBNA cytology comparing to histopathology was 97.17% and 98.32% respectively whereas specificity was 97.26% and 97.75 % respectively. Positive predictive value was 97.14% in BB and 99.66% in TBNA and negative predictive value was 93.23% in BB and 98.77% in TBNA. The accuracy of BB was 96.51% and 99.14% of TBNA cytology. Discordance of BB cytological and histopathological diagnosis was in 3.21%, whereas discordance of TBNA was in 2.03% cases. There was no statistically significant difference neither between BB (p=0.550) nor between TBNA (p=0.602) cytology and histopathological diagnosis. Conclusion: Cytology is valuable and useful in establishing lung cancer diagnosis, which yields almost the same information as histopathology no matter which method of cytological sampling is used.

Aim: To determine the degree of correlation between TTF-1 (+) expression and EGFR mutation status in lung adenocarcinoma. Introduction: Adenocarcinoma of the lung is mainly diagnosed based on standard morphological criteria. The thyroid gland transcription factor (TTF-1) is currently the most commonly used immunohistochemical marker in the differentiation of invasive adenocarcinoma of the lung from another primary and metastatic carcinoma, has a prognostic significance and is a predictor of the EGFR mutation status. Material and Methods: This retrospective study enrolled 60 patients with histologically confirmed primary lung adenocarcinoma who underwent lung cancer surgery at Institute for lung disease Vojvodina between 2010 and 2015. Tumor specimens of these patients were investigated for TTF-1 expression and mutations in EGFR using immunohistochemistry and PCR analysis. Statistical analysis is in statistic software Statistica 12. Results: The study included 35 men and 25 women, with an average age of 61.8 ą 8.08 years. Of the 60 cases, TTF-1 ( ) expression was recorded in 52 (87%) (p <0.001), the statistical difference is not significant when comparing smoking habitsby gender, and tumor size among them. EGFR ( ) mutation status was found in 3/60 (5%) cases [egzon 21 (2) and exon 20 (1)], of which TTF-1 (+) expression was in two cases. Conclusion: There is a statistically significant difference between the TTF-1 (-) and TTF-1 (+) adenocarcinoma and a high degree of correlation between EGFR mutation status and TTF-1 (+) expression.

Aim: We present the case of a woman with endobronchial pulmonary synovial sarcoma. Introduction: Primary pulmonary synovial sarcoma is an extremely rare tumor that has the same histomorphological characteristics and chromosomal translocations as the synovial sarcoma of soft tissue origin. Material and Methods: A woman aged 58 years, the smoker, without the current symptoms of the disease, came to our institution because of the nodus that was seen on the CT chest. The polip (26 mm) was located in the lumen of the lower right lobar bronchus. The mediastinal and hilar lymph nodes were not increased. The radiological examination was done as part of a routine control after the meningeoma surgery three years ago. Right lower lobectomy and resection of regional lymph nodes were performed. Results: By a macroscopic examination, in the lumen of the bronchus for the lower right lobes, a clearly limited, nonencapsulated, grayish-white node of 2.6 x 2.6 x 1 cm was found. Histologically, the tumor was showed interweaving fascicular uniform spindle cell with ovoid, pale staining nuclei, and inconspicuous nucleoli, scant cytoplasm and the cell borders indistinct. Immunohistochemical tumor cells were positive for CD99, bcl2 and vimentin. Surgical margins and regional lymph nodes were not affected. A detailed clinical and radiological examination confirmed the primary lung origin of the diagnosed synovial sarcoma. A year after surgery the patient feels good. Conclusion: Morphological and immunohistochemical analysis with detailed clinical and radiological examination confirms the primary lung origin of synovial sarcoma

Aim: To evaluate surgical lung biopsies in patients with a clinically and radiologically set diagnosis of ILD. Introduction: Interstitial lung diseases (ILDs) are a group of lung diseases affecting the lung interstitium. These entities share similar clinical and radiological features and are distinguished primarily by the histopathologic patterns on surgical lung biopsy. Material and Methods: The study included 30 patients with a surgical lung biopsy performed in 10-year period at the Institute for Pulmonary Diseases of Vojvodina in Sremska Kamenica. Standard H E stain, special stains for conective tissue and smooth muscle, as well as immunohistochemistry in some cases were used. The patient’s age, sex, clinical symptoms, surgical biopsy type and histological findings were analyzed. Results: Of the 30 patients who underwent surgical lung biopsy, an open lung biopsy according to Claassen was performed in 14 patients, in 12 biopsies biopsy according to Maassen was obtained, while in 4 patients material for histopathological analysis was taken by VATS (Video - Assisted Thoracoscopic Surgery). The most common biopsy site was upper lobe in 16 cases, then lingula in 10, middle lobe in 2, and lower lobe and lung base in 1 patient. By histopathological analysis, diagnosis of UIP in 8, PLCH in 7, sarcoidosis in 6, hypersensitivity pneumonitis in 3, NSIP in 2, LAM, LIP, DIP and ACIF in 1 patient. Conclusion: Diagnosis of ILD is based on history, physical examination, high-resolution CT imaging, pulmonary function tests, and lung biopsy which presents golden standard in diagnostic approach.

Aim: Analysis of two cases of IPA with an emphasis on the radiological and pathohistological findings of this entity. Introduction: Aspergillus spp. can cause a wide range of lung diseases, depending on the current state of immunity and the existing pulmonary diseases. Invasive pulmonary aspergillosis (IPA) is severe form of pulmonary mycosis, with the appearance of granulomatous inflammation with the development of necrosis and suppuration, as well as the invasion of hyphae into pulmonary parenchyma and the blood vessels and spreading the disease out of the lungs. Material and Methods: In the five-year period, two cases of IPA were diagnosed at the Institute of Pulmonary Diseases of Vojvodina. Material for pathohistological analysis, obtained by surgical method and on autopsy, was stained with standard H E staining, as well as with special staining methods: PAS and Grocott. Results: Patients were 67 and 48 years old and both were treated for acute lymphoblastic leukemia. They were admitted to our hospital in respiratory insufficiency and severe neutropenia with a radiologically diagnosed IPA based on HRCT finding of “halo sign”. This sign pathohistologically corresponds to foci of necrosis of lung parenchyma surrounded with the zone of hemorrhage. In addition to these foci of necrosis, in the wall and lumen of blood vessels, numerous septate hyphae with dichotomous branching at 45° were found. Conclusion: Although the pathohistological diagnosis is golden standard for diagnosis of IPA, given the invasiveness of the techniques for obtaining material for analysis, diagnosis can be made based on HRCT finding of “halo sign”.

The significance of pathohistological analysis of operative material and tumor immunophenotype in differential diagnosis of sarcomatoid carcinoma and carcinosarcoma rare malignant lung neoplasms. Introduction: Lung carcinosarcoma, biphasic tumor, composed of non-small caracinomatous and sarcomatous heterogeneous components, is a rare neoplasm and represents less than 1% of all malignant lung tumors. Case report: A 64-year old man reported to physician complainig of fever that lasted for two months. Chest CT showed signs of right lung infiltration, close to the hilus and mediastinal lymphadenopathy. After bronchoscopy and biopsy of the suspected changes pathohistological finding was: carcinoma squamosum invasivum. Lobectomy of upper right lobe and mediastinal lymphadenectomy were performed. On gross examination, distanced 5 mm from the resection margin, withish tumor (22x7x5mm) infiltrating the wall of the lobar bronch was found. Histological analysis found that tumor tissue was partly consisted of atypical epithelial cells, with larger, mitotically active nuclei with focuses of keratinization and partialy of atypical oval and spindle cells,with hyperchromatic, pleomorphic nuclei, with high mitotic index. Lymph nodes were negative for tumor tissue. Results of immunohistochemical tumor tissue analysis: Cytokeratin - positive in carcinomatous component, Vimentin - positive in sarcomatous component, Actin, Desmin and S100 - negative. According to the results of morphological and immunohistochemical analysis of the tumor tissue lung carcinomsarcoma was diagnosed. Conclusion: For establishing definitive pathohistological diagnosis in biphasic tumor cases, a pathohistological examination of operative material is necessary, and morphological assessment should be suported by immunohistochemical analysis.

Aim: To investigate immunohistochemical expression and the localization of connexin-43 (Cx-43) in primary lung cancer and its metastases. Introduction: Connexins are transmembrane proteins forming gap junctions that allow intercellular communication. Significance of gap junctions and connexins in lung cancer are not clear enough. Material and Methods: We analyzed autopsy samples of primary and metastatic lung carcinoma from Institute of Pathology in Belgrade. There were 11 primary lung carcinomas, 7 lung cancer metastases in lymph nodes, and 12 haematogenic metastases. We performed immunohistochemical staining for connexin-43 (Cx43) and measured expression (percentage of positive cells and intensity of staining) and localization of Cx43 in primary tumor and its metastases. Results: Lymphatic and hematogenous metastases of lung cancer showed a stronger expression of connexin-43 than primary tumor itself. Unlike 9% of primary carcinoma, 28% of lymphatic metastases and 50% of hematogenous metastases had expression of connexins in more than 50% of tumor cells (p=0.11). The intensity of connexin-43 expression was statistically significantly less in primary lung cancer than in all the metastases together(p=0.04). The expression of this marker was different in different histological types, where small cell carcinoma rarely expressed connexin, while the squamous carcinoma was mostly positive to immunohistochemical staining on Cx43. Dominant localization of expression was the combined cytoplasmic-membranous. Conclusion: Our results showed that lung cancer expresses connexin-43 mostly in cytoplasm as well as on the cell membrane. Further research on a larger sample is required to establish whether Cx-43 could be used as a prognostic biomarker in lung cancer.

Aim: We present a case of a patient with pneumothorax and subcutaneous emphysema as the first manifestation of miliary tuberculosis. Introduction: Miliary tuberculosis is the result of hematogenous dissemination of Mycobacterium tuberculosis in patients with weak immuno-defensive mechanisms. Pneumothorax and subcutaneous emphysema are possible complications of miliary tuberculosis. Case report: A woman aged 64 years old reported to the regional institution because of breathing difficulties. On the radiograph of the chest, pneumothorax was observed left, and the left thoracic drain was placed. Subcutaneous emphysema and global respiratory insufficiency were reported an hour later after which the patient was transferred to our facility. At the admission the patient was in poor general condition, intubated, hemodynamically unstable, markers of inflammation were elevated with the presence of electrolyte imbalance and severe anemia. On the chest radiogram, there was recorded: pneumothorax left, pneumonia right and generalized subcutaneous emphysema, and thoracal drain that was placed. Intensive therapy had improved the condition of the patient, after which she was extubated. Progression of respiratory insufficiency and lethal outcome occurred on the second day of admission. An autopsy was performed. A macroscopic examination and pathohistological analysis found: massive subcutaneous emphysema in the chest, well-placed thoracal drain, bilateral pleural effusion, bilateral acute tuberculous caverns in the lungs and necrotizing granulomas in: the lungs, liver, spleen and larynx which have led to asphyxiation and aviation outcome. Conclusion: In poorly-fed patients with the development of pneumothorax, subcutaneous emphysema and severe respiratory disorders, it is necessary to suspect tuberculosis.

Aim: To examine the quality of tested lung carcinoma samples, frequency and type of EGFR mutations, and their correlation with patients clinical characteristics (gender, age, smoking habits, clinical stage). Introduction: Mutations in Epidermal Growth Factor Receptor (EGFR) have a role in lung carcinoma development and they are more prevalent in women and non-smokers. Evaluation of EGFR mutations in lung carcinomas in mandatory for targeted therapy with tyrosine kinase inhibitors. Test performance depends on the quality of tested samples and a test type. Material and Methods: We evaluated reports of EGFR mutation real-time PCR analyses in lung carcinoma samples performed from June 2017 till February 2018. Presence of mutations was correlated with clinical characteristics of lung carcinoma patients. Results: A total of 341 samples was received for testing, among which 40 (11.7%) was unsuitable for analysis due to a low tumor cell content (<5%). Three types of mutations were detected in a total of 24 (8%) cases: L858R in 12 (50%) cases, exon 19 deletion in 10 (41.7%) cases, and G719A/C/S in two cases (8.3%). Mutations were more prevalent in women (13.7%) then in men (4.3%) (p=0.004). Patients with EGFR mutated tumors were older (67,6ą9,4 years), compared to those with non-mutated tumors (62,3ą8,8 years) (p=0,003). Smoking habits and clinical stage were not associated with mutation status in lung carcinomas. Mutations were detected only in adenocarcinomas. Conclusion: Our results suggest the low frequency of EGFR mutations in tested patients, but they are more prevalent in women and older patients.

Aim: The aim of this research is to analyze the profile of Ezh2 expression in superficial urothelial bladder cancer, to investigate its correlation with clinicopathological parameters, as well as to determine the prognostic significance of Ezh2. Introduction: Superficial urothelial bladder cancer, without invasion of muscle layer, is associated with frequent recurrence, and represents significant burden for health care system. Ezh2 is epigenetic regulator with a major role in urothelial oncogenesis. Clinical investigations of Ezh2 inhibitors in treatment of solid cancers have already given encouraging results. Materials and Methods: Tumor samples from 410 patients with superficial bladder cancer (172 pTa, 238 pT1), obtained by transurethral resection, were incorporated in tissue microarrays, and analyzed immunohistochemically to Ezh2 expression. Correlation analysis with clinicopathological parameters was performed using SPSS 18.0. Results: High nuclear expression was found in 33.4% tumors, and it was significantly more frequent in pT1 (46.6%), compared to pTa tumors (15.1%) (p<0.001). Ezh2 expression was associated with high histologic grade, presence of carcinoma in situ, and cancer specific death (p<0.001, respectively). In Kaplan-Mayer survival analysis high Ezh2 expression was significantly associated with poor prognosis and shorter patients survival (p<0.001). There was no significant correlation between Ezh2 and recurrence of the disease, and recurrence free interval (p<0.05). Conclusion: Immunohistochemical expression of transcription repressor Ezh2 in superficial urothelial bladder cancer indicates aggressive behavior of the tumor, and poor prognosis. Ezh2 could be used as pronostic marker in selection of the patients that might require more intense clinical treatment, and as potential target of anticancer therapy.

Aim: Case report for rare complication rectorrhagia induced by rectal lipoma incarcerated in the anus . Introduction: Colorectal lipomas are rare tumors that are commonly diagnosed in the right colon, accidentaly during colonoscopy. When the lipomas are larger then 2 cm, they cause pain, bleeding, obstruction, incarceration and torsion. Material and Methods: We present the case of 50-year old man who comes to emergency ambulance with abundant rectorrhagia and blood presented on underwear and thighs. It is noted prolapse of the soft structure through the anus which is reponated into the anus. Anoproctoscopy was performed, which determines that it is polyp of rectum, although it seemed to be incarcerated hemorrhoids, due to the fact that the patient has been suffering from hemorrhoids with bleeding for several years,which is treated conservatively. It was found that it was not hemorrhoids prolaps or bleeding from them. Flexibile rectoscopy was performed on the untreated gut. The polypoid structure on peduncle,was verified in the distal rectum,3,5 cm from the pectinate line. Polypoid formation was electroresected and sent for pathohistological examination. Results: The patient was well tolerated intervention. Resected specimen revealed sessile pseudopolypoid tumor,eroded mucosa , diameter 28x25x24 mm.Histopathology revealed submucosal lipoma . Eroded mucosa is accompanied by focuses microbloods. Microcircuits of fatty necrosis are visible inside the lipoma. Conclusion: Lipom of the rectum is rare entity which is accidentaly diagnosed during colonoscopy. Extremly rare, lipom causes bleeding, which we present here.

Aim: To reported extremely rare case of collision adenocarcinomma et small cell neuroendocrine carcinomma of the gallbladder (SCNEC). Introduction: Collision cancers are malignancies in the same organ or anatomical site that comprises et least two different tumor components, with no mixed or transitional area between two component. Case report: 76 year old woman with abdominal pain, underwent ultrasonography evaluation which demonstrated cholelithiasis and gallbladder wall thickening. Cholecystectomy due to cholelithiasis was performed.The macroscopic analysis revealed 2,5cm sized round nodular lesions in the fundus of the gallbladder.Formalin fixed, paraffin embedded tissues were stained with H.E. Selected samples were stained immunochistochemically with chromogranin, synaptophysin et CK7. Microscopicaly, the tumor was composed of two components. Dominant component is adenocarcinomma, composed of tubular glands lined predominantly by columnar cells with pseudostratified et ovoid or elongated nuclei.In the area close to this component there was neuroendocrine carcinomma that came in touch with the previous one, but didnt infiltrate it. Neuroendocrine carcinomma was composed of round or fusiform cells, arranged in sheets, nests and cords.Tumor cells have round hyperchromatic nuclei with inconspicuous nucleoli. Neuroendocrine tumor cells were immunoreactive for chromogranin, synaptophysin. Epithelial cells were positive for CH7.The final pathologycal diagnosis was SCNEC. The tumor stage was II, T2, Nx, Mx. Conclusion: Prognosis for patient is poor.About 40-50 percent of patients have disseminated disease at the time of the diagnoses.SCNEC appear to be highly responsive to chemotherapy as well as radiotherapy and survival time more than one year have been reported.

Aim: We present the case of a rare localization of the alveolar soft part of the sarcoma in the visceral organ. Introduction: Alveolar soft part sarcoma (ASPS) is a rare mesenchymal tumor typically occurring in young patients, more frequently in females. Common localization of ASPS is skeletal musculature of lower extremities. ASPS in visceral organs usually represents a metastasis from the more common primary location in skeletal muscles. ASPS is characterized by a tumor-specific translocation which causes the fusion of the TEF3 with a ASPL gene (also known as ASPSCR1). Case report: Female 47 years old was admitted to hospital due to abdominal pain. Urgent surgery was performed due to ileus. Ileal tumor was detected intraoperatively as a cause of ileus. Tumor was infiltrated whole intestinal circumference, with dimension 70mm x 47cm and evident perforation. Histology showed well-defined nests of pleomorphic cells separated by delicate fibrovascular septae. Within described nests there is a prominent lack of cellular cohesion, representing for the distinctive pseudoalveolar pattern. Immunohistochemical stadies were diffusely positive for TFE3 and focally positive for CD34 and alpha-SMA and negative for panCK, DOG-1, CD117, S-100, HMB45, Desmin. Immunopositivity for Ki67 was present in 20% of tumor cells. FISH analysis was done using locus specific dual color break-apart TFE3 (3 and 5 ) probe and rearrangement in the TFE3 gene was confirmed. Conclusion: Despite the fact that ASPS is rare mesenchymal tumor in visceral organs it have to be considered as possible diagnosis especially in cases with typical histological features and immunohistochemical profile. Definitive diagnosis of ASPS must be confirmed by FISH analysis.

Aim: We present a rare case of gastrointestinal tumor in a boy. Introduction: Gastrointestinal stromal tumors (GISTs) represent rare mesenchymal tumors in pediatric population. Most of these tumors originate in the stomach and very rarely in the oesophagus. They are more frequent in young girls. As opposed to adult GISTs, those that occur in childhood show mutations in PDGFRA receptors or so called “Wild type” mutations. Case report: A fifteen years and 2 months old boy with symptoms of dysphagia and stenocardia for the last six months. Endoscopy showed nodular solid lesion at the distal part of the thoracal oesophagus, later surgically removed. Dimensions of the lession were 35x25x20mm with whitishPOSTER SESIJA 57 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. yellow cut surface and areas of hemorrhage. Histology showed predominant spindle cell pattern with minor epithelioid component, with pronounced cellular and nuclear pleomorphism, rare areas of necrosis and hemorrhage, with the mitotic count more than 18 per 50 high power fields and numerous atypical mitotic figures. Immunohistochemical studies were CD117 and DOG-1 negative, but positive for PDGFRA. Tumor expressed focal alpha-SMA and CD34 positivity. Conclusion: Informations regarding pediatric gastrointestinal stromal tumors are still insufficient. Even though these tumors are rare in childhood, when analysing mesenchymal neoplasms potential GISTs should be considered.

Aim: To present a case report of choristoma of the stomach. Introduction: Ectopic pancreas is defined as the presence of pancreatic tissue outside its normal location without anatomic or vascular connections with the pancreas. It mostly occurs in upper gastrointestinal tract, predominantly in the stomach. It s likely to occur in men, at the age of 30-50 years. Case report: A thirty-year old male patient presented to the hospital for evaluation of epigastric symptoms. After clinical examinations, he underwent an esophago-gastroduodenoscopy which showed intramural tumor located along the greater curvature. The biopsy was taken which revealed normal gastric mucosa so it was suspected for gastrointestinal stromal tumor. The patient was submitted to laparoscopic partial gastrectomy and the material was received for pathohistological analysis. Macroscopically, there were several fragments, gray-pink coloured and lined with pink mucosa. In the largest fragment it was noticed irregular, oval gray-white node, measured 4x2.5x0.5cm. On serial cutting it was yellow. Histologically, described fragments revealed normal fundic mucosa and irregular node was consisted of pancreatic tissue localized in submucosa and lamina muscularis. It was formed of normal acini and Langerhans islets, with focally dilated pancreatic ducts and presence of mixed inflammatory infiltrate. Dysplasia wasn t found in the pancreatic tissue. The POSTER SESIJA 58 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. established diagnosis was Ectopic pancreatic tissue in stomach Choristoma of the stomach. Conclusion: Choristoma is rare condition and usually incidental finding important to be diagnosed because of the serious complications depending on which tissue is present in the organ, such as inflammation, bleeding and mailignant transformation.

Aim: We present an interesting case of pancreatic ectopic tissue in the gallbladder. Introduction: Ectopic or heterotopic pancreas is defined as the presence of pancreatic tissue outside the boundaries of the pancreas that show no anatomical or vascular connection with the main body of the pancreas. Case report: We present a rare case of ectopic pancreas found in a 38 year-old man s gallbladder. Male patient was admitted to the Surgical Department Clinical Hospital Center Zvezdara, as scheduled for an elective laparoscopic cholecystectomy. One year prior to the surgery he had had abdominal ultrasonography done during a routine hospital check up. Ultrasonographic examination of the whole abdomen had showed no abnormality, except for cholelithiasis. Laparoscopic cholecystectomy was done. On gross examination, the gallbladder measured 9 cm in length and 3.5 cm in circumference, with a wall thickness ranging from 0.2 to 0.4 cm. On cutting open, one yellowish round stone, measuring 0.6 cm in diameter, was noted in the fundus. The mucosa was velvety flattened. A nodule of 1,5 cm in diameter was seen in the neck region, which on microscopic examination, showed a well circumscribed rest of heterotopic pancreatic tissue, composed of lobules of exocrine pancreatic acini and an occasional duct. Islets of Langerhans were also present. Conclusion: Ectopic pancreatic tissue in a gallbladder is a very rare condition which is usually diagnosed incidentally. Up to the presents, only about 30 cases have been reported. The clinical significance of the ectopic pancreas remains unclear and it requires further exploration.

Aim: Pathohistological diagnostics (PD) of the Yolc Sac tumor (YST) of primary mediastinal localization. Introduction: YST are malignant germ cell tumors of primitive endodermal gonadal differentiation. In 1% -5% of cases their localization is extragonadal, including mediastinal in 50% -70% published cases. Case report: A 30-year-old man was admitted to the Thoracic Surgery Clinic for diagnosis and treatment of mediastinal tumor with lungs and liver metastases. Laboratory analyzes revealed high elevations in serum alpha-fetoprotein (AFP), in addition moderate elevation of beta-chorionic gonadotropin (ß-HCG). No pathological changes in testis and retroperitoneum have been found. An open biopsy of the mediastinal change was made for PD. Histologically, tumor tissue showed a significant degree of necrosis. There were reticular, microcystic and pseudopapillary forms of neoplastic cell growth, surrounded by myxomatosis stroma. Tumor cells cytoplasm was scarce and vacuolated with high nuclear polymorphism and hyperchromasia. Positive immunohistochemical (IMH) reaction was obtained for panCK, AFP, PLAP. Immunoreactivity for TTF-1 was found in about 50% of tumor cells, as well as the focal reaction for ß-HCG in rare multinuclear cells. Reactions for D2-40, CD5, CK-7, CK-20, p63, CD117, CD30, napsin A and calretinin were negative. Conclusion: The results of our analysis showed the expression of divergent endodermal linear markers in the mediastinal YST, especially TTF-1 expression. The diagnosis of YST in small biopsies can be difficult and requires using of a wide range of IMH markers in order to closer the differentiation of primary tumor localization and the application of appropriate chemotherapy

Aim: Reporting of autopsy case of fatal accidental poisoning by colchicine from meadow saffron. Introduction: Meadow saffron (Colchicum autumnale) is a perennial herbaceous plant similar to the edible wild garlic (Allium ursinum). Toxic substance in meadow saffron is alcaloid colchicine. Colchicine poisoning is a very dangerous condition, which can lead to fatal outcome. Case report: A 50-years-old male was admitted to the hospital complaining of weakness, abdominal pain, nausea, vomiting and diarrhea without blood. Day before he ate two plants thinking they’re wild garlic and seven hours after ingestion he felt first symptoms. During the course of the hospital stay, he had gastroenterocolitis, acute renal failure, hepatic lesion and cardiorespiratory insufficiency with fatal outcome. Post-mortem examination reveled:brain oedema, lung oedema and congestion, heart weighing 700 grams, ventricular hypertrophy, myocardial fibrosis, liver congestion and steatosis, spleen congestion, pancreatic fibrosis. Organs sections were taken for pathohistological analysis. Body fluids and parts of organs were toxicologically analyzed. Pathohistological findings:brain oedema, diffuse perivascular and interstitial myocardial fibrosis, myocardial haemorrhage, lungs congestion and edema, micro- and macrovesicular liver steatosis, centrilobular liver necrosis, lymphocytic inflammatory infiltrate in liver portions, spleen’s red pulp congestion, kidney congestion and interstitial bleeding, coagulation necrosis of the proximal tubules of the kidney. Toxicological analysis showed colchicine in blood-0.011 mg/L, urine-0.051 mg/L, liver with gallbladder-0.007 mg/ kg, kidney-0.008 mg/kg. Conclusion: Ingestion of the meadow saffron can lead to poisoning with fatal outcome due to colchicine. Colchicine intoxication should be suspected in patients with gastrointestinal symptoms after consuming wild plants.

Aim: The present a rare case intraocular primary malignant choroidal melanoma. Introduction: Melanoma choroideae is most common intraocular primary tumor of adults. Incidence is 7 cases per 1 million people per year and incidence of this tumor increases with age. Case report: We report 76 years old women who had progressive visual field defects at left eye at least two years. Diagnostic examination of eye was detected a greyish-brown mass of the posterior choroidal pole which was indication for enucleation. Macroscopically bulbus oculi was dimension 28x25x25mm, with optic nerve 5mm length. At the intersection on posterior pole nearby papila of optical nerve was detected ovoid dark lesion diameters 8x3mm. Histopathology examination was showed sharply demarcated solid tumor islands which consists of epithelioid tumor cells with pleomorphic, atypical nuclei, conspicuous nucleoli. Other type of cells were spindle look which formed fascicular shapes. Pigmentation was absent also. There was no significant vascularisation in tumor mass. Sclera was superficially infiltrated by tumor cells. Mitotic activity was absent (0/40HPHF). Immunohistochemistry staining was confirmed melanoma cells: S-100, MelanA , HMB-45 /-. Conclusion: Melanoma in this region is rare, indolent lesion, with bad prognosis. Early detection could prevent further dissemination of disease and many complications. Metastatic choroid melanoma is a highly malignant disease with a limited life expectancy.

Aim: The aim of this case was a correct diagnosis of mediastinal tumor in a 41-years old female patient. Introduction: The rarity of primary extraneural ependymomas, its great variations in morphology and rare occurrence of metastasis, increase chances of misdiagnosis. Case report: Macroscopic examination of received specimen was performed, followed by histological and immunohistochemical analysis of the tissue samples. In presented case, onset of the disease was 14 years ago, when after right salpingo-oophorectomy, patient was diagnosed with malignant mesothelioma. In following years patient had multiple and extensive surgical procedures, resulting in different patohistological diagnosis, and after seven years, a diagnosis of extraneural ependymoma was established. Later on, patient was surgically treated in several medical centers across the region, again with different patohistological diagnosis. At present, tumor metastasized to mediastinum, presenting as grey to brown, multicystic formation, with cysts filed with clear serous fluid or red-brown hemorrhagic fluid. Inner surface of the cysts had smooth to partly papillary appearance. Tumor cells exhibited several architectural paterns (solid, pseudorosette or rosette formations, papillary and pseudopapilary structures), and immunophenotype specific for extraneural ependymoma (GFAP, ER, PR positive, calretinin, WT-1, S100, synaptophysin, chromogranin, CK7 and pan-cytokeratin negative). Conclusion: This case demonstrates an important principle in tumor pathology. Neoplasms may occur in unusual and unexpected primary and metastatic sites. Pathologists need to be familiar with histologic features of a wide range of neoplasms and not just the appearance of neoplasms within their own limited subspecialty area.

Aim: We present a case wherein the information obtained from autopsy examination was of critical importance to a doctor and a family. Introduction: A 24-year-old,multipara,delivered a term born male baby with a birth weight 3100gr and body length 46cm.Soon after birth the neonate developed signs of POSTER SESIJA 63 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. a respiratory insufficiency and died within 2 hours.Anamnestic data from the mother revealed uncontrolled pregnancy. Material and Methods: Standard autopsy technic and standard procedure of paraffin embedded section routinely stained with H E was performed. Results: At autopsy,the external examination revealed characteristic facial features suggestive of Potter s face including posterior rotated low-set ears,flat nose,widely separated eyes,micrognatia and short neck. Autopsy revealed presence of bilateral hypoplastic lungs with total weigh of 28g,less than the expected range (normal 49g).On dissection lungs were airless, non-crepitant and sank in the water. Histologically findings were consistent with a diagnosis of pulmonary hypoplasia.The abdominal cavity was completely filled with markedly symmetrical enlarged kidneys.Total weight of both kidneys was 156g (normal 25g).On the dissection section showed multiple small cysts measuring 1-3 mm in size,completely replacing the cortex and medulla giving it a spongy like appearance and typical honeycomb structure.On microscopical examination we found cysts uniformly lined by cuboidal to flattened epithelium. Zaključak: We consider that this is a Potter Syndrome Type I due to Autosomal Recessive Polycystic Kidney Disease and is linked to a mutation in the gene PKHD1.2. Through this case,we are aware of the importance and benefit of autopsy,although the trend of autopsies in the world is decreasing.

Aim: Determining the immunohistochemical characteristic of parathyroid glands (PG) proliferative activity in patients with primary and secondary hyperparathyroidism (HPT) using cell cycle and proliferation immunohistochemical markers, Ki -67 and PCNA. Introduction: A few studies results have shown A few studies results have shown significant detection of Ki-67 in hyperplasia due to secondary hyperparathyroidism (sHPT), whereas it s demonstrated only in adenomas in primary HPT (pHPT). The highest PCNA expression is detected in hyperplastic PG in sHPT and in adenoma in pHTP, but in pHTP hyperplasia it s extremely low. Material and Methods: We analyzed the surgically removed PG of 96 patients with HPT. In POSTER SESIJA 64 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. addition to standard histopathological parameters the results of immunohistochemical reaction of Ki-67 and PCNA in 23 normal, 73 hyperplastic PG and 23 adenoma were analyzed. Results: 41 (42.7%) patients had pHPT, and 55 (57.3%) sHPT. Within pHPT adenoma was diagnosed in 23 (56.1%) patients and hyperplasia in 18 (43.9%). Detection of PCNA was 94.4% in pHPT hyperplasia, 91% in sHPT hyperplasia, and 83% in adenoma. 22 (98%) of the normal PG didn t have PCNA expression. The expression of Ki-67 was found in 13 (56.5%) adenomas and in 11 (18.3%) nodular hyperplasia. The high statistical significance for Ki-67 (p <0.0001) was found between adenoma and pHPT and sHPT. Conclusion: The results of our analysis showed high Ki-67 and PCNA expression in parathyroid adenomas. Increased Ki-67 expression corresponds with increased cellular proliferation and contributes to tumorigenesis in many organs, but doesn t distinguish accurately benign from malignant PG tumors.

Aim: We present an interesting case of unexpected metastatic deposit “signet ring cell” of the gastric cancer in the ureter. Introduction: Ureteral obstruction caused by gastric cancer may occur by any of the following three mechanisms: direct extension from the primary site, peritoneal deposit or lymph node metastasis. Material and Methods: We report the case of a patient with ureteral metastasis as the first and sole manifestation of gastric cancer dissemination two years after he was first diagnosed with gastric cancer. Results: A 52-year old male patient was admitted to the Department of Urology, Clinical Hospital Centre Zvezdara in Belgrade, with a 5-day history of right colic flank pain and high temperature. He had a partial gastrectomy for gastric cancer two years ago and received 8 cycles of chemotherapy. Routine blood test results were normal except elevated creatinine and C-reactive protein levels. An abdominal ultrasound examination and computed tomography revealed grade 3 hydronephrosis and hydroureter. Cystoscopy indicated a tumor measuring 2 cm in size which involved left ureteral orifice. A left nephroureterectomy and transurethral resection of bladder tumor were performed. Histopathological examination of surgical POSTER SESIJA 65 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. specimen revealed signet ring cell carcinoma infiltrating lamina propria and tunica muscularis of the left lower ureter. Histopathological examination of the bladder specimen revealed signet ring cell carcinoma identical to those of the ureteral tumor. Conclusion: Ureteral metastases from gastric cancer are extremely rare. Shimoyama reviewed 27 cases of the true ureteral metastasis from gastric cancer. The prognosis is generally poor and the survival for more than 2 years has not been reported in literature.

Aim: Reporting a patient with unusual metastatic site of invasive lobular breast cancer (ILC) as initial presentation of the disease. Introduction: Due to specific growth pattern, ILC rarely forms an apparent tumor, which makes diagnosis very challenging at early stage. ILC is also known for unconventional metastatic spread, with deposits being discovered prior to the primary tumor in 3-10% of cases. Case report: While evaluating renal function in 51-year old female patient hospitalised at the Urology Clinic (Clinical centre of Montenegro), static scintigraphy revealed left kidney functional capacity of 7-8%. Nephrectomy was indicated. Kidney, 11x6x4cm in size, with slightly reduced, paler parenchyma, firmly attached fatty capsule and pyelocaliceal system and ureter of regular gross appearence, was delivered to the Centre for Pathology. Analysis of H E sections revealed chronic pyelonephritis. In a few sections taken from urether, pyelon and subcapsular parts of parenchyma, infiltrates of small, cuboid, atipical cells, mostly arranged in one-cell-thick files, were noted. Immunohistochemistry reveiled strong pozitivity for EMA, CK(ae1/ae3), CK7, estrogen and mammaglobin, with Ki67<10%. A few cells were progesteron positive, while vimentin, CK20 and neuroendocrine markers were negative. ILC metastasis was suspected. ILC, with axillary lymph POSTER SESIJA 66 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. node involvement, was confirmed later, although there was no macroscopically apparent tumor in the breast. Tumor cells were estrogen and progesterone positive, HER2 negative, with Ki67 of 3%. Conclusion: While assessing metastatic deposits in unconventional sites in women, primary ILC should be considered. Special diagnostic algorhytm is required for efficient initial detection of the primary tumor.

Aim: Immunohistochemistry findings along with clinical features, are significantly important in differentiating the Extrapleural SFT in the neck, from other well-circumscribed mesenchymal neoplasms at this locations. Introduction: We present a rare case of a Extrapleural SFT in a 57 years old man in the neck, without significant past medical history. Material and Methods: The patient had a painless slow growing tumor, in right sight of the neck, diagnosed with physical examination. Total excision with local anesthesia was done, without previously biopsy of the tumor and other clinical investigations. Standard procedures for histology and immunohistochemical stains were done. Results: Tumor was well circumscribed, encapsulated measuring 5,5x4x4 cm. On section, the cut surface had a multinodular, whitish and firm appearance. On microscopic examination tumor was composed of alternating hypocellular and hypercellular areas separated from each other by thick bands of collagen and branching haemangiopericitoma like vessels. The tumor cells were round to spindle-shaped with little cytoplasm, indistinct borders, dispersed chromatin within vesicular nuclei. Area of myxoid change and subcapsular focus of hemorrhage was present, and 2 mitoses/10 HPF were found. Immunohistochemistry revealed diffuse positivity for CD34, Vimentin and BCL2, focal positivity for CD99, S100, SMA, and negativity for CKWS and EMA. Ki67 showed low proliferating index 3-5%. Conclusion: Although most cases of SFT are benign, there is no strict correlation between morphology and behavior, so patients with extrapleural solitary fibrous tumor have need of long-term post-resection follow-up. Further studies are needed to determine the optimal management of these neoplasms.

Aim: To present three cases and review literature of splenic myoid angioendothelioma (SMA) focusing on immunohistochemical features. Introduction: SMAs are very rare, mainly in middle-aged and elderly patients of both sexes and are characterized by a mixed proliferation of cord capillaries and myoid cells, distinguished from other splenic angioendotheliomas by additional myogenic/myofibroblast differentiation. Material and Methods: Three cases of SMA from the Department of Histopathology Registry of Clinical Centre of Serbia were detected during last 12 years (2006-2017) in one male and two female patients (42,5 ys average age). Histomorphological findings were revised by reviewing all serial HE sections, histochemical trichrome stains and immunohistochemical stainings for CD8, CD31, CD34, CD68, SMA, desmin and Ki-67. Results: All cases showed sharply demarcated non-encapsulated solitary tumors with diameters 42, 55 and 20 mm. Histologically there are dense network of capillary blood vessels intermingled with polygonal myoid stromal cells and at least focally expressed non-homogeneous cellularity of stromal and lymphoid cells with focal sclerosis. Neocapillaries show distinctive CD8- / CD31 / CD34 immunophenotype (differing them from splenic hamartomas) and characteristic mixture with myogenic elements (differing them from cord capillary hemangiomas): intense SMA (3/3), rare focal desmin (2/3) and focal CD68 (1/3) immunoexpression. Conclusion: SMA is underrecognized type of vascular neoplasia, which has a clinico-pathological differential diagnostic significance because it radiologically imitates splenic metastase. Cellular form of SMA must be distinguished from hamartoma, but also from hemangiopericytoma and well-differentiated angiosarcoma of the spleen

Aim: Purpose of this report is to present metastasis of lymph nodes melanoma with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) as an example of collision tumor of an uncommon synchronic occurrence in the lymph node. Introduction: Synchronic, composite tumors are rare and their simultaneous and synchronic occurence within the same tissue/organ is even more rare. CLL/SLL is an indolent, clonal disease of mature B-lymphocytes which occurs mostly with adults. After the treatment with chemotherapeutic agents the occurence of the secondary malignancies (melanoma,squamous-cell carcinoma). Material and Methods: We present a 77 year old male who, after right sided nephrectomy caused by clear-cell carcinoma was diagnosed with CLL/SLL with bone marrow and lymph node infiltration. After three years and chemotherapy, skin changes were excised which histomorphologically resembles melanoma. After the immunotherapy for melanoma, the enlarged lymph nodes were extripated from the neck and histomorphologically and immunohistochemically treated. Results: Histomorphologically, a diffused infiltration of small B-lymphocytes was found in the lymph node, with round nuclei, condensed chromatin, inconspicuous nucleoli, scant cytoplasm unique immunophenotype: CD20 ,PAX-5 ,CD5 ,CD23 ,Cyclin D1-,CD3-. A part of lymph node was infiltrated by epitheloid cells with immunohistochemic profile: S-100 ,Melan A ,HMB-45 . Histomorphological and immunohistochemical CLL/SLL with melanoma infiltration as an example of a collision tumor was proved. Conclusion: Lymphoproliferative neoplasms including CLL/SLL represent an risk of synchronous, metachronous development of secondary malignancies including melanoma itself and they can uncommonly present themselves as synchronic collision tumors within the same organ.

Aim: We present the case of a patient aged 68 years who died of chronic heart failure caused by untreated cardiac sarcoidosis (CS). Introduction: Sarcoidosis is a multisystem disorder of unknown etiology, characterized by granulomatous infiltration and the development of noncaseating granulomas in many organ systems. Although the lungs, eyes, and skin are most commonly affected, virtually any body organ can be involved. Clinical evidence of CS is seen only in 5% of patients and the disease may present with tachyarrhythmias, conduction disturbance, heart failure, or sudden cardiac death. Case report: The patient was received in the hospital due to symptoms and signs of global cardiac decompensation with difficulties in the form of dyspnea, orthopnea, and edematous legs. Echocardiographic, cardiac cavities are very dilated with globally reduced systolic function, severe mitral regurgitation and ejection fraction about 20%. Very soon after receiving the patient is died. At autopsy, the heart was dilated, primarily the left ventricle. Histologically, the myocardium was infiltrated by numerous granulomas built of lymphocytes, epitheloid cells, and giant multicellular cells of the Langhans type. As a consequence of severe chronic heart failure, the lungs were edematous with both sides of massive plural effusions. The coronary arteries were non-significantly stenosed. Conclusion: Early diagnosis and treatment can prevent significant morbidity and mortality in patients with CS. It is very important that patients with CS in the early stage of the disease be treated with corticosteroids.

Aim: To analyze age and sex distribution of the cardiac tumors, the most common clinical symptoms, POSTER SESIJA 70 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. pathohistological types, tumor localization and compare the echocardiographic with the pathohistological size of the tumor. Introduction: The incidence of primary cardiac tumors are very rare and amounts to about 0.02%. Incidence of secondary cardiac tumors are significantly higher. Primary malignant cardiac tumors are very rare and represent to the rarest tumors in the human organism. Materials and Methods: The study covered 49 patients who were operated in the period from 2008 to 2017. Patients data are obtained from the history of the disease, the information system and pathohistological findings. Results: The average age of the patients is 53.9 years. The most common symptoms were light fatigue, heart palpitations, vertigo, dizziness, dyspnea, exhaustion, cough and stenocardia. The most commonly diagnosed are myxomas (67.3%), papillary fibroelastomas 28.6%, cavernous hemangiomas 4.1%, and metastatic tumors 2.04%. The most common tumor localization is in the left atrium 63.3%, aortic cusps 16.3%, right atrium 8.2%, mitral valve 8.2%, left ventricle 2.04% and interventricular septum 2.04%. The difference in mean echocardiographic tumor size and tumor size after surgical extirpation was not statistically significant (p = 0.706). Conclusion: Although, cardiac tumors are rare, they have a large clinical importance, primarily because of the potential for severe complications such as embolization of the arteries of the brain with the development of cerebral infarction and even the appearance of sudden death. However, timely diagnosis and surgical removal of tumors lead to patient curing in most cases.

Aim: We present an illustrative autopsy case of thrombosed giant bilateral vertebral artery aneurysms. Introduction: A 61-year-old male died at Department of Infective disease with a clinical diagnosis for bilateral bronchopneumonia, cerebral aneurism, fibro muscular dysplasia, CVI, HTA, chronic CMP, cardiac arrest. Material and Methods: Standard autopsy technic with neuropathology brain dissection and standard procedure of paraffin embedded section routinely stained with HE was performed. Results: Gross examination of the brain at the autopsy showed saccular giant bilateral vertebral artery aneurysms that measured 58 mm on the right and 40 mm on the left artery. They were tumor-like and compressed the medulla and pons. Both were thrombosed. The right was ruptured with subacute subarachnoid bleeding. Sections of the cerebral vessels exhibited minimal atherosclerotic plaques, with mild stenosis (focally up to 10%) of the left internal carotid artery. We found mild dilatation on ventricles and minimal cortical atrophy of the brain. During the microscopic examination angiodysplasia with abnormally dilated blood vessels on visceral organs, predominantly on brain and heart was detected. The cause of death was central type of cardiopulmonary insufficiency with pulmonary edema. Conclusion: We presented an extremely rare case with bilateral giant vertebral aneurysms. Giant cerebral aneurysms are ones that measure >25 mm in greatest dimension and account for ~5% of all intracranial aneurysms. They occur in the 5th-7th decades and are more common in females. Vertebral artery aneurysms constitute 0.5 to 3% of intracranial aneurysms and 20% of posterior circulation aneurysms.

To present pathological processes of the TG with histological detection of granulomas, analysis of morphological forms of granulomas, and their diagnostic significance. This paper is based on literature review and insight into the archival materials of the Institute of Pathology and Forensic Medicine of the Military Medical Academy.The presence of granulomas in the thyroid gland (TG) includes specific pathological processes such as subacute thyroiditis (SAT) and palpation thyroiditis (PT). The clinical manifestations of the granulomas may be accompanied by symmetrical or asymmetrical enlargement and palpatory pain in the gland, which requires further clinical examination. Granulomas in the TG can be associated with various benign and malignant processes. There are two large groups of granulomas: foreign-body giant cell granulomas (FBG) and immune granulomas (IGR). FBG are histiocytic reactions to chemically inert, exogenous or endogenous materials. Etiologically, IGRs arise in the framework of infectious, autoimmune, toxic, drug-induced or pathological processes of unknown etiology. According to the presence of necrosis IGRs can be further divided as necrotizing or non-necrotizing type. TG granulomas of the infectious, autoimmune or inflammatory nature of the unknown etiology are extremely rare. 1. Granulomas in specific pathological processes of the TG Subacute (de Quervain’s) thyroiditis or granulomatous thyroiditis is an inflammatory process that is clinically presented as enlarged and painful TG. In most cases, the result is a complete recovery of the TG function. Permanent hypothyroidism is found in about 5% of patients. SAT is usually preceded by upper respiratory tract infection. The disease is etiologically related to viral infections, genetic predisposition and the use of immuno therapy. Macroscopically, TG is usually symmetrically enlarged, but there are also localized forms with nodular morphology, which imitate neoplastic lesions. The microscopic characteristic is the presence of multifocal and diffusely distributed folliculocentric granulomas. They are found in different phases and consist of epitheloid histiocytes, lymphocytes, plasma cells, neutrophils, and multinuclear giant cells (MGC). At the center of the granuloma, the colloid is reduced or absent. In later phases, fibrosis can develop perifollicularly. In terms of differential diagnosis (DDG), it is important to differentiate SAT from other granulomatous inflammations. Palpation thyroiditis (Multifocal granulomatous folliculitis) is the most common pathological process in TG with microscopic detection of granulomas. It is an incidental microscopic finding involving individual or minor follicular groups. Changes arise as a result of mechanical microtrauma after the palpation of the TG. Microscopic changes are characterized by damage to the follicles with interfollicular APSTRAKTI 73 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. accumulation mainly of histiocytes in the presence of lymphocytes, plasma cells and MGCs. In the DDG of PT, the following conditions must be considered: SAT, primary and secondary microscopic foci of papillary microcarcinoma, C-cell hyperplasia, and focal forms of Langerhans histiocytosis. 2. Foreign-body giant cell granuloma is frequent incidental microscopic finding in TG. It arises as a reaction to the accumulation of endogenous substances in the areas of spontaneous or degenerations induces by fine-needle aspiration biopsy (FNAB). The most common forms of FBGs on endogenous material are cholesterol granulomas. These FBGs are composed of MGCs, foamy histiocytes, and hemosiderophages arranged around crystal deposits. Depending on how old the lesion is, there may be a focal necrosis, a different degree of fibrosis, extracellular deposits of hemosiderin, and other inflammatory cells. The presence of FBGs and histiocytic aggregates is not only important in the preoperative cytological diagnostics, but also in the post-operative pathohistological analysis of TG nodules. Large nuclei of histiocytes with hypochromasia, nuclear membrane irregularities and the presence of MGC can imitate the cytological features of papillary thyroid carcinoma (PTC). Exogenous biomaterials are rarely cause of FBGs in TG. After thyroidectomy, in cases of diagnosed TG malignancies, the presence of suture FBGs in thyroid bed imitates recurrence or the rest of malignancy and is the cause of repeated surgeries. 3. Necrotizing granulomas (NGR) in TG Granulomas with necrosis may be of infectious and noninfectious etiology. Tuberculosis is the most common cause of NGR in TG. Tuberculosis in TG can be presented as a solitary nodal lesion, diffuse microlesions, nodular goiter, and rarely as an abscess or a chronic skin sinus. As a infectious cause of NGR in TG, sporadically reported cases have been caused by histoplasmosis, coccidioidomycosis and nocardiosis. Rare non-infectious NGR in TG or in the TG bed, of autoimmune etiologies, have been described as part of Wegener’s granulomatosis and rheumatoid arthritis. Post-operative necrotizing granulomas also represent NGR of non-infectious cause. Microscopically, there is a morphology that matches post biopsy granulomas in other organs (prostate, urinary bladder). 4. Non-necrotizing granulomas (NNGR) in TG Sarcoidosis is a multi-systemic chronic granulomatous inflammation of unknown etiology. Thyroid is rarely affected by sarcoidosis. Macroscopically, the gland is diffusely or nodularly enlarged or reduced in volume. Interstitially localized NNGR represent a typical histological presentation. Sarcoidosis of TG should be distinguished from the sarcoid-like stromal reactions of PTC in the gland or regional lymph nodes. In these cases, it is necessary to clinically exclude the systemic disease. 5. Granulomas and histiocytic reactions in neoplastic processes of TG Apart from the described FBGs, PT and sarcoid-like reactions, in epithelial tumors of the TG histiocytic aggregates (not granulomas) may also be seen as secondary changes after FNAB. Interfollicular/ intraluminal presence of MGCs with or without the presence of histiocytes and granuloma-like morphology represents a characteristic finding in PTC. The cytological and histological detection of MGCs is one of the diagnostic criteria for PTC. Their presence in tumors may be due to a reaction to an altered colloid produced by PTC or as a non-specific immune response to due tumor cells. Conclusion: Granulomas in the TG are not rare. Knowing the morphology of granulomas, pathological processes and the circumstances in which they occur is significant in DDG of primary tumors of the TG, their recurrence and metastases in the cervical lymph nodes. The diagnosis of granulomatous inflammation in TG can be based on the histological characteristics of granulomas in correlation with clinical and laboratory findings.

Ova mala kliničko-patološka kategorija tumora se odlikuje nepredvidljivošću kliničkog toka i/ili ishoda bolesti, a tome u prilog često govore i patohistološka obeležja tumora koja pružaju samo neke odlike maligniteta, sugerišu mogući maligni potencijal ili pokazuju odlike između benignih i malignih proliferacija1. Boljim definisanjem dijagnostičkih kriterijuma danas su ovi tumori smanjeni na manje od 5% svih tumora u GI i HBP sistemima, a njihov maligni potencijal se često posredno izražava kao proliferativni, recidivni ili metastatski potencijal2,3. Najveći broj njih odnosi se na dobro diferentovane neuroendokrine tumore (NET), gastrointestinalne stromalne tumore (GIST) i mucinozne cistične neoplazije (MCN). Od epitelnih neoplazija nejasnog malignog potencijala u digestivnoj cevi se posebno izdvajaju mucinozne neoplazije apendiksa i cekuma (do 0,2% tumora GIT) često praćene rupturom i peritonealnim pseudomiksomom kao tzv. „diseminovana peritonealna mucinoza“4. One se odlikuju „cistadenomskom“ morfologijom, ponekad kompleksne acinusne i mikrocistične organizacije sa displastičnim epitelom i mucinoznom (pseudo)invazijom zida, ali bez invazivnosti epitelnih elemenata5. Slična histološka obeležja vide se u kod mucinoznih cističnih neoplazija (MCN) u biliopankreatičnom traktu i jetri, gde se displastične epitelne promene duktusa pankreasa (analogne pankreasnoj intraepitelnoj neoplaziji PanIN I-III), i bilijarnih duktusa (analogne bilijarnoj intraepitelnoj neoplaziji BilIN I-III) u perifernim zonama razgranavanja moraju jasno razlikovati od rane invazije, tj. od adenokarcinoma6. Kad su u pitanju adenomatozni polipi sa pseudoinvazijom u digestivnom traktu, oni se moraju razlikovati od pravih „malignih polipa“ tj. adenomatoznih polipa sa ranom invazijom koji pokazuju disekciju stromalnih elemenata mukoze i submukoze i izazivaju dezmoplastičnu reakciju. Pseudoinvazija se odlikuje odsustvom infiltrativnog tipa rasta, dezmoplazije, uočavanjem strome (lamine proprije) oko prolabiranih glandularnih struktura, hemoragijom, hemosiderofagijom, a nekada i invertnim rastom, bilo pulzionim ili limfovaskularnim, kao i stvaranjem limfoglandularnih kompleksa sa fokalnim limfoidnim agregatima7. Kada su u pitanju dobro diferentovani NET, danas se pored klasične i imunohistohemijske dijagnostike obavezno moraju odrediti i korelirati mitotski indeks (MI) izražen na 10 polja velikog mikroskopskog uveličanja, ali određen na najmanje 50 polja, kao i procentualni proliferativni Ki-67 indeks određen u području najveće aktivnosti na najmanje 500 ćelija. Na taj način određuje se stepen maligniteta NET koji po novoj klasifikaciji iz 2017. godine koji pored NET-G1 i NET-G2 podrazumeva i novouvedeni dobro diferentovani NET visokog stepena (NET-G3) za tumore sa MI preko 20 /10HPF i Ki-67 indeks preko 20%, ali koji se moraju razlikovati od slabo diferentovanih neuroendokrinih karcinoma (NEC) istih ili viših vrednosti ovih parametara8. Značajna je i grupa visceralnih mezenhimalnih tumora nejasnog malignog potencijala koji se pojavljuju u digestivnom sistemu, posebno za GIST kao najčešći od njih i koji čini oko 2% klinički značajnih tumora u abdomenu. Danas su jasno definisani AFIP kriterijumi malignog potencijala (Miettinen, 2005) i NIH kriterijumi metastaskog potencijala (Fletcher, 2002) koji čine i sastavni deo aktuelnih TNM klasifikacija. Oni uključuju obavezne podatke o preciznoj lokalizaciji i veličini tumora i mitotskom indeksu izraženom na 50 polja velikog mikroskopskog uveličanja9. Na osnovu toga se prepostavljaju stepeni rizika malignog toka bolesti i uključivanja adjuvantnih terapijskih protokola, dok je za lečenje metastatske i/ili neresektabilne bolesti neophodno određivanje mutacionog statusa KIT, PDGFRA, SDHA i SDHB gena. Takođe je značajno prepoznavanje i nalčin dijagnostike inflamatornog miofibroblastnog tumora (IMT) koji najčešće zahvata adolescente i mlade ženske odrasle osobe (75% intra-abdominalno i ponekad multinodularno), a histološki se odlikuje karakterističnom mešavinom inflamatornih (posebno limfoplazmocitnih i histiocitnih) i stromalnih (posebno miofibroblastnih) elemenata10. Karakteristična nuklearna imunoreaktivnost za ALK-1 protein se vidi samo u oko 50% slučajeva. Ovaj tip tumora ima nepredvidiv tok, ali se recidivi (29%) i metastaze (4%) češće javljaju kod epiteloidnih hipercelularnih i mitotski aktivnijih tumora sa pojavom nuklearne atipije stromalnih ćelija11. Hepato-biliopankreatični tumori nejasnog malignog potencijala osim pomenutih MCN podrazumevaju i intraduktalne papilarne mucinozne neoplazme (IPMN) sa teškom epitelnom displazijom (analogne BilIN III, PanIN III) ali bez pridružene mikroinvazije ili jasnih adenokarcinoma12. Takođe, hepatomi nejasnog malignog potencijala su hepatoidni tumori koji odgovaraju hepatocelularnim adenomima (HCA) s ati- 75 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. pijom neoplastičnih ćelija i koji ne ispunjavaju sve ostale parametre maligniteta (elementi invazivnosti, trabekularna dezorganizacija i povećanje broja gredica, evidentna mitotska aktivnost) a po pravilu su “beta-katenin aktivišući tip” HCA koji se imunohistohemijski može dokazati i koji pokazuje visok stepen maligne transformacije13. Usled toga je neophodno ekstezivno uzorkovanje velikih HCA tumora da se ne bi previdela fokalna maligna alteracija. U jetri se retko može videti i epiteloidni hemangioendoteliom (EHE), nepredvidivog kliničkog toka, koji se često pogrešno interpretira kao (adeno)karcinom. Po pravilu se javlja kod osoba srednjeg životnog doba, kao multipla nodularna promena (80%) i javlja se s ekstrahepatičnim širenjem u 50% slučajeva. Neophodno je da se kod mladih jasno razlikuje od infantilnog hemangioendotelioma. Mikroskopski se odlikuje zonalnošću sa centralnom miksohondroidnom i sklerotičnom promenom u kojoj se vide dendritične tumorske ćelije, dok se periferno uočavaju delom obliterisani a delom prošireni sinusoidi sa papilarnim projekcijama atipičnog endotela14. Često se u tumorskim ćelijama vide intracitoplazmatske vakuole koje mogu sadržati eritrocite. Mora se razlikovati od dobro diferentovanog angiosarkoma visokom celularnošću i jačom atipijom, što je povezano i sa malignom alteracijom EHE. Pankreasna solidno-pseudopapilarna neoplazija se odlikuje češćim benignim kliničkim tokom nego recidivima i veoma retkim metastaziranjem. Opisala ju je Virginia Franz (1959) kao pedijatrijski benigni tumor, ali se danas izdvaja boljom mikroskopskom karakterizacijom kod adolescenata i mladih (10-45 god.). Obilno se javlja u distalnom pankreasu, promera od 3-15 cm kao jasno ograničen, lobuliran, hemoragičan tumor. Histološki ga odlikuju papilarnost, pseudorozete, holegranulomi i fokalna hijalinizacija15. Maligni potencijal i rizik metastaziranja se povezuju sa hipercelularnošću, atipijom i izraženijom mitotskom aktivnošću. 

Case 1: Clinical History -The patient was an 18-year-old woman with a history of primary amenorrhea. The physical examination revealed normal female external genitalia, moderate breast development, scarce pubic hair, and scant axillary hair. Pelvic examination revealed an absence of a uterus. The vagina ended in a blind pouch and measured 6cm in length. An abdominal ultrasonogram showed solid bilateral gonadal structures measuring 31x15mm right, and 30x 6mm left in the pelvic cavity without any evidence of a uterus. These findings were confirmed by MRI scan. Serum levels of estradiol and testosterone were 45.7pg/mL and 7.5nmol/L, respectively. FSH and LH levels were 8.46mlU/ml and 20.2mlU/ml. Cytogenetic analysis of peripheral blood lymphocytes revealed a 46,XY karyotype, with translocation t(13,14) inherited from her father. The molecular investigation confirmed active SRY with amplification of 6 nonpolymorphic loci on AZFa, AZFb, and AZFc regions on Y chromosome. Three months after her first referral the patient underwent bilateral prophylactic laparoscopic gonadectomy. The patient is being followed up regularly and she has no complaints after five years. Pathological Findings- Grossly, the right gonad was 4.3x1.8x1.8cm and the left gonad was 4.5x2.0x1.7cm. The right fallopian tube was 3.8cm long and 0.2cm in diameter, while the left one was 1.5cm long and 0.2cm in diameter. In addition, 4 cysts measuring 0.3- 1cm in diameter were found in the fat tissue on the surface of the left fallopian tube. The cut surface of the right gonad resembled testicular tissue showing two discrete, firm, well-demarcated, slightly bulging, homogenous, light grey-tan colored nodules, measuring 0.1 and 0.9cm in maximal dimension. Three similarly looking nodules were present in the left gonad measuring 0.4-0.5cm in diameter. In addition, a white whorled, firm, smooth muscle body fused to one pole of the gonad was found bilaterally measuring 1.2cm right, and 1.3cm left. Microscopically, both gonads were composed of varying proportions of small and solid, or less frequently larger, with early lumen formation seminiferous tubules lined by immature Sertoli cells, surrounded with fibrous, focally edematous stroma in which prominent Leydig cells were present. Rare tubules contained Sertoli cells with abundant cytoplasm filled with coarse, eosinophilic granules. The nodules were composed of solid immature tubules lined by cylindrical immature Sertoli cells separated by fibrous stroma containing fewer Leydig cells. Fallopian tubes were hypoplasic, while the cysts were lined by a single layer of cuboidal to columnar cells some of which had cilia. Thus, the clinical, laboratory, imaging, genetic and histological findings confirmed the diagnosis of complete androgen insensitivity syndrome with bilateral testicular hamartomas.Discussion Androgen insensitivity syndrome (AIS) is a disorder where there is resistance to androgen actions influencing both the morphogenesis and differentiation of androgen-responsive body structures. It is the most common type of male pseudohermaphroditism, characterized by an absence of androgen receptor activity due to a mutation at Xq11–q12 localization on the androgen receptor gene. the. In the largest series of 43 patients with the AIS published by Rutgers and Scully hamartomas were present in 63% of the cases, while Sertoli cell adenomas were reported in 23% and malignant tumors including two seminomas, one intratubular germ cell neoplasm with early stromal invasion and a malignant sex cord tumor in 9% of the cases. Case 2: Clinical History- The patient was a 17-year-old girl with primary amenorrhea. On external examination, she had an unambiguous female phenotype albeit with poor breast development. Her external genitalia had a normal appearance but internal examination showed a hypoplastic uterus. Ultrasonography failed to show follicular activity in the gonads. Her gonadotropins were high (FSH, 55.6mU/ml; LH, 18.3mU/ml) and estradiol was low (20.0pg/ml). Nonmosaic 46,XY karyotype was detected in patient’s leukocytes by cytogenetic analysis. Under the tentative clinical diagnosis of pure gonadal dysgenesis, a prophylactic bilateral laparoscopic gonadectomy with bilateral salpingectomy was performed. A biopsy specimens from the gonads were sent for cytogenetic analysis. The sequencing of the SRY gene of the proband revealed a C/G substitution at the first nucleotide of codon 133, leading to Arg/Gly replacement in the SRY protein. The mutation was also present in patient’s father, who is a phenotypically normal male. APSTRAKTI 77 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. The patient is alive and well 12 years after the operation without any evidence of disease. Pathological Findings- Macroscopically, both fallopian tubes were 4cm long and 0.4cm in diameter, while the gonads were measuring 3x2.2x1.2cm the right, and 2.2x0.8x1cm the left. Upon dissection, both gonadal streaks had a similar, variegated appearance with multiple areas of microcalcification. The histological evaluation of the gonads confirmed a gonadoblastoma of 3cm in diameter in the right gonad and a predominantly “burnt-out” gonadoblastoma of 2.2cm in the left gonad with only a microscopic focus of recognizable gonadoblastoma less than 1mm in diameter. The tumors were composed of primordial germ cells intimately admixed with sex cord elements, which surrounded round spaces filled with eosinophilic basement membrane-like material. The atypical germ cells had abundant clear or pale, slightly granular cytoplasm and round vesicular nuclei with prominent nucleoli. The small, often fusiform sex cord element cells had uniform round or oval nuclei with indiscernible nucleoli and scant cytoplasm. The uninvolved gonadal tissue bilaterally had the morphology of a streak gonad composed largely of ovarian-type stroma, with extended calcification and small germinal inclusion cysts present, without any follicles. The fallopian tubes were infantile. Therefore, the histopathological findings confirmed the diagnosis of bilateral gonadoblastoma in streak gonads, FIGO Stage IB, in a patient with a XY gonadal dysgenesis (Swyer syndrome). Discussion: XY gonadal dysgenesis (GD) is a result of abnormal testis development in utero. Pure 46,XY GD should be considered when an adolescent presents as a phenotypic female with delayed puberty and primary amenorrhea. Rarely, they may present with a detectable abdominal or pelvic gonadoblastoma mass. When the presumptive diagnosis suggests GD, further criteria may strengthen the diagnosis, while ultimately, the most definitive diagnosis is through biopsy of the gonads. Bilateral streak gonads are seen in pure XY GD. The risk of otherwise undetected gonadoblastoma in XY GD patients is high, and prophylactic or therapeutic gonadectomy is therefore often indicated when XY GD is diagnosed. 

Sarcomatoid differentiation in renal cell carcinoma (sarcomatoid renal cell carcinoma – sRCC) represents a dedifferentiated high grade neoplasm of the kidney that contains carcinomatous and sarcomatous component. sRCC is an uncommon tumor, with the incidence of 5-8% of renal carcinoma. Sarcomatoid differentiation is a result of divergent differentiation of malignant epithelial cells. During this process tumor cells lose their epithelial features, and acquire mesenchymal characteristics, which provides them higher APSTRAKTI 79 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. capacity for migration and metastasis.This phenotype can be encountered in all RCC subtypes, including clear cell, papillary, chromophobe RCC, and carcinomaof collecting duct. Sarcomatoid component may resemble fibrosarcoma or undifferentiated pleomorphic sarcoma. Less often heterologous differentiation can be found, with histologic patterns resembling rhabdomyosarcoma, chondrosarcoma, or osteosarcoma. Rhabdoid phenotype is an aggressive form of divergent differentiation which has been described in clear cell RCC, papillary RCC, as well as in chromophobe RCC (CRRCC). CRRCC comprise about 6% of RCC, and has better prognosis compared to other RCC subtypes. Sarcomatoid differentiation is found in 8% of these tumors, and is an indicator of poor prognosis, as it is in other RCC types. Sarcomatoid component may represent terminally dedifferentiated clone which can arise from any RCC subtype, or can develop from a special clone. Although sRCC is most frequently found in high grade tumors, the occurrence of sarcomatoid differentiation in RCC, Fuhrman 1/2, detected in 30% of the cases, denies the claim that sarcomatoid differentiation is a continual process of classic RCC dedifferentiation, but a consequence of sarcomatoid stem cell activation within the tumor. In a case of sarcomatoid differentiation in renal carcinoma it is necessary to detect focuses of any morphological type of renal cell carcinoma, and to verify the immunohistochemical positivity of sarcomatoid component to epithelial and mesenchymal markers. Their epithelial origin is confirmed by the expression of cytokeratins, and EMA. PAX8, CAIX, and CD10 can be used to confirm the renal origin, which is of particular importance in finding of sarcomatoid cells in needle biopsy, as well as in metastatic RCC. CAIX, p53, and Bcl-2 can play a major role in transformation of RCC to highly malignant neoplasm with sarcomatoid differentiation. The expression of CAIX, p53, and Bcl-2 is indicator of poor prognosis and worse survival of the patients with sRCC. Significant micromorphological parameters that may influence the clinical course of the disease comprise percentage of sarcomatoid differentiation (>50%), presence of vascular invasion, extent of necrosis, and advanced TNM stage. Type and grade of sarcomatoid component, as well as RCC subtype do not influence the clinical outcome. The prognosis of patients with sRCC is worse compared to prognosis of other histologic forms of RCC. Sarcomatoid differentiation in RCC is a significant cause of mortality, with average survival of 4-9 months after diagnosis. The majority of unclassified RCCs contains variable percent of tumor cells with sarcomatoid morphology, without recognizable epithelial component. About 3% of RCC are exclusively sarcomatoid. These tumors are larger, more often invade adrenal glands and other adjacent organs, metastasize more frequently into regional lymph nodes and bones, and have significantly shorter survival (average is 4.3 months). Differential diagnosis of sRCC should exclude primary renal sarcoma, sarcomatoid urothelial carcinoma, and angiomyolipoma. Micromorphologically sRCC may resemble classic sarcoma, however it should be taken into account that primary renal sarcomas are extremely rare in adults, and comprise less than 1% of renal malignancies. The half of primary renal sarcomas are leiomyosarcomas, consisting of smooth muscle component, which is extremely rare in sRCC. In sarcomatoid urothelial carcinoma, the location of tumor is of significance, also the presence of carcinoma in situ, recurrence of urothelial carcinoma, and immunohistochemical positivity to CKhmw and p63. In angiomyolipoma, immunoreactivity to HMB45, as well as the presence of focuses with classic morphology, excludes sarcomatoid RCC. Conclusion: Sarcomatoid differentiation in renal cell carcinoma is a result of divergent differentiation of malignant epithelial cell, and can be found in all renal cell carcinoma subtypes. During this process tumor cells lose their epithelial features, and acquire mesenchymal characteristics, which provides them higher capacity for migration and metastasis. It is very important to recognize the presence of sarcomatoid differentiation in renal cell carcinoma, since it is an indicator of poor prognosis irrespective of RCC subtype.

Introduction: Hamartomas of the bile duct named von Meyenburg complex are benign liver lesions consisting of dilated bile duct structures with a surrounding fibrous stroma. Their incidence is age-dependent and they are observed about 1% in children and 5%-6% in adults. Von Meyenburg complexes are infrequently observed lesions, characterized by multiple small nodular lesions located below the Glisson’s capsule, and ranging from 0.1 to 1.0 centimeters in diameter. Von Meyenburg complex of the liver are usually detected during laparotomy or autopsies an incidental finding. Multilocular occurrence is possible although they are rarely spread throughout the whole liver, as it was observed in our patient. They are normally asymptomatic, and are incidental findings in asymptomatic patients. They may be found in normal liver tissue, but also in association with Caroli’s syndrome, congenital hepatic fibrosis, autosomal dominant polycystic renal diseas, cholangiocarcinomas and cholangitis. Cholangiocarcinoma which arise from these lesions are usually lower stage and better differentiations than other type of cholangiocarcinoma. The sonographic findings of von Meyenburg complex are variable, including multiple, small, hyperechogenic images, with poorly delimited margins, or even hypoechogenic images with a “target” pattern with a hyperechogenic center and a hypoechogenic periphery, and well delimited margins. A magnetic resonance cholangiography is the best imaging examination of hamartomas of the bile duct, which can distinguish the different forms of dilatation of the bile duct. Histology of von Meyenburg complexes consists of a variable number of dilated small bile ducts, embedded in a fibrous, sometimes hyalinizing stroma. Microscopically, they are characterized by cystic dilatations of the bile duct or clusters of mature bile duct of various sizes, peri-ductal glands, and encompassed by fibrous stroma. The ductules are lined by small cuboidal or flattened cells, with round to oval nuclei. Bile duct hamartomas contain cysts that are more irregularly shaped then normal ducts, and they may also contain eosinophilic debris or inspissated bile. Case report: A 68-year-old male patient with multiple hepatic lesion which ultrasonic and MSCT appearance suggestive of multiple liver metastases was accepted for surgical exploration and liver biopsy. The patient had one mounts symptoms of vomits and weight loss. During surgery numerous whitish irregular lesions of various sizes scattered in the hepatic surface imitating metastatic deposits were noted trough both liver lobe and trough all liver quadrants. Explorations of the rest of abdominal cavity not found any pathological changes or peritoneal carcinomatosis. Liver biopsy was done and taken three samples for analysis. Tissue was brown-yellow-gray color and medium-firm consistency. Histological analysis demonstrated multiple lesions composed of biliary ducts incorporated in fibrotic tissue (Figure 1).There are usually cystic dilatations of some intrahepatic biliary ducts with irregular shape lined with uniform epithelium (Figure 2). The epithel of biliary ducts in von Meyenburg complex were immunohistochemicaly Epithelial Membrane Antigen positive, Pan-Cytokeratin positive, Cytokeratin 7 negative, Cytokeratin 5/6 negative and Carcinoembryonic antigene negative. Also present were signs of cholestasis with small lakes of bile.Conclusion: Von Meyenburg complexes are an important differential diagnosis of liver metastases. Differential diagnosis of liver metastases also includes other benign liver lesions, including hemangiomas, adenomas or infectious lesions e.g. miliary tuberculosis. As the existence of liver metastases is crucial for therapeutic decision making in malignant diseases, this differential diagnosis must be carefully clarified. Since VMC are usually less than 5 mm in size, they can escape preoperative radiologic diagnostics. The macroscopic appearance of von Meyenburg complexes can mimic liver metastasis as demonstrated in our reported patients.

Introduction: Pancreatic lesions, made of spindle cells, are a heterogeneous group of lesions, ranging from reactive, inflammatory changes to tumors. Differentiation of an individual lesion is difficult and requires the use of additional analytical methods (histochemical, immunohistochemical and molecular), and a comparison of morphological characteristics with other characteristics of the changes (radiologic and laboratory characteristics). We will present two cases of benign spindle cell lesions of the pancreas, with reference to the differential diagnosis. Material and Metods: The first patient was a female, aged 51 years, with a change localized in the pancreatic head, diameter of 9.5 cm. The second patient was a male, aged 35 years, with a change in the pancreatic tail, with maximum diameter of 5.5 cm. Results: In a female patient, the lesion was an inflammatory myofibroblastic pancreatic tumor, built of fascicles of mostly spindle cells (fibroblasts/myofibroblasts). The cells had uniform, elongated, spindle nuclei and eosinophilic cytoplasm. They were arranged in short fascicles that occasionally made storiformn formations. Mitotic activity of spindle cells was low (0 - 2 mitosis/ HPF 10, FD 0.65). In the stroma, there was a mixed inflammatory infiltrate, consisting of lymphocytes, plasma cells, histiocytes, eosinophils and neutrophils. In between, there were fascicles of collagen, together with the parts of the pancreas (excretory ducts, lobules, acini, and parts of the endocrine pancreas) (Figure 1). Immunohistochemically, spindle cells showed a diffuse immunohistochemical positivity to: Vimentin, SMA and Desmin. Negative immunohistochemical reaction was showed to S-100, p53, CDX2 and ALK-1.

Aim: The aim of this study is to present a case of benign lymphadenitis that cytologically was interpreted as suspicios for malignant lymphoma. Introduction: This study represents a case of a patient with cervical lymph node swelling in which the cyto-diagnosis performed by fine needle aspiration cytology was suspicious for lymphoma. The correct diagnosis was assessed by subsequent histology after the removal of the enlarged cervical lymph node. Material and Methods: For cytologic study the material was obtained by fine needle aspiration biopsy and syringe washings, air-dried smears and alcohol-fixed smears, which were prepared and appropriately stained by PAP and May Gruenwald-Giemsa stains. For correct diagnosis an extirpated lymph node was properly fixed and processed with routine haematoxylin eosin staining as well as with an additional immunohistochemical analyses. Results: The cyto-histologic features were characterized by a polymorphous population of cells, germinal center cells with large nuclei, a few epithelioid-type cells and histiocytes with intracellular inclusions The cytological diagnosis implied suspicion for malignant lymphoma probably of Hodgkin type. Histologic features revealed a reactive lymph node architecture that immunohistochemically revealed the diagnosis of Toxoplasma lymphadenitis. Serologic testing for toxoplasma in other institution revealed elevated titres that established the histopathological diagnosis. Conclusion: Lymphadenitis due to Toxoplasma infection is common and should be considered in the diagnosis of unexplained lymphadenopathy at all sites, especially the cervical region. Serologic confirmation should be recommended for all suspected cases and unlike in this case, fine needle aspiration cytodiagnosis can eliminate the need for hospitalization and surgery.

Aim: The aim of this paper is to point out the importance and the role of immunohistochemistry in diagnosing rare benign epithelial tumours of the lung and a very similar malignant tumour of well-differentiated lepidic adenocarcinoma. Introduction: In pulmonary pathology diagnostic dilemmas are frequent. One of the most complex challenges is to differentiate between benign tumours of pneumocytes and other forms of similar tumours. In particular, it is difficult to differentiate between the tumours of the same or similar histogenetic origin and morphological characteristics. However, dilemmas can also be related to whether a tumour has benign or malignant potential. In order to be able to have proper diagnostics, we need to have a detailed insight in the morphological and immunohistochemical features of these tumours. One of the best KRATKI KURSEVI APSTRAKTI 85 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. examples of this are two very rare and morphologically very similar benign epithelial tumours: sclerosing pneumocytoma (according to the 2015 World Health Organisation Classification of Lung Tumours; new terms changed or entities added since 2004; the 2004 World Health Organisation Classification called it sclerosing haemangioma)1 and alveolar adenoma on the one hand; and well-differentiated lepidic adenocarcinoma on the other hand. These are most often cited as the most problematic in terms of their differential diagnostics. When it comes to first two tumours, as it can be concluded from their original names, they were considered to be the tumours of completely different histogenetic origin. However, their immunohistochemical profile and all current data show that they have identical structure and origin. Immunohistochemical diagnostics enabled the demystification of neoplastic processes, as is the case with rare benign tumours of pneumocytes. This diagnostics can also point out the biological potential and help differentiate between benign and malign tumours. Additional dilemma is posed by the fact that sclerosing pneumocytoma may even give metastases into regional lymph nodes, which do not affect disease prognosis 2,3. Histopathological differential diagnosis includes, apart for the above mentioned, other benign epithelial tumors, hemangioma, primary and metastatic carcinoma4. Materials and methods: We analysed two very rare and morphologically very similar benign epithelial tumours, (sclerosing pneumocytoma and alveolar adenoma) and welldifferentiated lepidic adenocarcinoma. It was also performed their immunohistochemical analysis using the following markers: Cytokeratin 7 (CK7), Thyroid transcription factor 1 (TTF-1), Epithelial membrane antigen (EMA), Pan-cytokeratin (CK), Carcinoembryonic Antigen (CEA), FVIII, Ki67 and p53. Results: The first tumour, at the microscopic level, showed sclerosing and haemorrhagic arrangement, with ectatic spaces filled by blood and solid areas and papillary-like formations. Basic cell population was epithelial cells, dominantly with eosinophilic and partially with granular cytoplasm. The nucleus was in the centre, round, without prominent nucleoli and mitoses. Stroma was moderately pronounced and centrally it was denser and composed of bundles of oval and spindle-shaped fibroblasts. Some of the cavities within the tumour had wide, cavernous space, lined with endothelium-like attenuated cells. Mainly in the middle part of the tumour, we could see the areas of hyalinisation of connective tissue. The tumour borders were expansive. The tumour did not infiltrate the pleura. On the final histopathological slides, the second tumour had a microcystic appearance. In central parts there was pale amorphous, homogenous content. Spaces were lined with cylindrical cells containing acidophilic and clear cytoplasm. Stroma was scarcely developed and sometimes with more pronounced parts and composed of groups of elongated spindle-like fibrocytes and fibroblasts. Immunohistochemical analysis of both tumours showed very similar reactivity: Ck7, TTF-1, EMA and CK showed diffuse positivity, k67 showed low proliferation index <1%. Cea in the major part of sclerosing pneumocytoma was negative and focally individual cells had reactivity, while alveolar adenoma was negative in its entirety. P53 and FVIII in both cases showed negative results. After all analyses, the definitive diagnosis of the first tumour is pneumocytoma and for the second one alveolar adenoma. The third tumour showed similar morphology as the previous two. At the microscopic hematoxylin eosin stain, it was dominantly composed of alveolar-adenoid formations. Tumour cells were bulky, cubic or polygonal; foamy, pale acidophilic, with homogenous cytoplasm and hyperchromatic roundish nuclei without prominent nucleoli. The immunohistochemical analysis of the third tumour showed positive reactivity with Ck7, TTF-1, CK, Cea, EMA, k67 proliferation index > 32%, while p53 proliferation index ≥1%, while the FVIII had a negative result. Final diagnosis for this tumour is well-differentiated lepidic adenocarcinoma. Conclusion: Due to almost identical histopathological and immunohistochemical characteristic, there may be a diagnostic dilemma: are these two separate tumours or this is the same tumour. Taking into consideration that sclerosing pneumocytoma give positive epithelial immunohistochemical reaction, their earlier name is wrong. Previous examples are good indicators of how we should adapt the names of tumours to their real nature and this is a good recommendation in terms of how we should organise future classifications. All of the above mentioned points to the fact that with these tumours it is necessary to have immunohistochemical evaluation and that we have to introduce new immunohistochemical predictive and prognostic markers. It is necessary to determine the cut off values for proliferative markers.

The skin is the largest organ in our body.Receiving information from the environment allows the role barrier between the human organism and the environment. The histological structure of the skin is variable depending on the part of the organism. The diseases of skin also vary depending on the region. The reason is the local immune status and the external physical, chemical and microbiological environment. The histopathological diagnosis of skin lesions request detailed clinical information. On the other side the histopathological diagnosis of skin lesions provides information on local disease , as well as potential other associated pathological conditions.˝ Extramammary Paget‘s vulvar disease is rare. The pathology of the skin of the vulva is specific both for specific localization, as well as for specific friction and microbiological flora. It is clinically presented as erythematosus or eczematous lesion. It can be local or extensive. Histopathologically, it is characterized by relatively large glandular atypical Paget cells, which are pathognomonic for this disease. These cells have abundant cytoplasm that is granular or vacuolated. Tumor cells are typically localized individually or in groups in the basal and parabasal layers. Immunochemical analyzes are necessary in the diagnosis of Paget‘s disease. The main reason is that Paget‘s disease can be a primary skin disease or associated with non-cutaneous carcinoma primarily of the urothelial or rectal carcinoma . Therefore, it is necessary to recommend that a detailed clinical trial of a patient be conducted in all diagnosed cases of vulvar Paget‘s disease. Mycobacterium marinum is etiological factor of Fish tank granuloma.This Mycobacterium more often causes diseases of fish, both marine and river fish. The humans become infected with Mycobacterium marinuim after contact with skin after micro/trauma Infection is usually localized to the skin.In immunocompromised patients the infection may disseminate or spread to the subcutis and bone. After incubation of few weeks after infection the lesion appear as solitary nodules or plaques.In some cases the disease progresses like suppurative ulcers. Histopathologically there is a wider range of histopathological presentations. Most often you can see abscess, necrosis of wheat and fatty tissue. However, granuloma inflammation is a key morphological substrate. After granuloma inflammation, fibrosis occurs. Since the microscopic image may be somewhat non-specific, clinical data are of great importance for the diagnosis. Multidisciplinarity in the final diagnosis includes microbiological confirmation of the presence of Mycobacterium marinum.

Introduction: A pleural effusion describes an excess of fluid in the pleural cavity, usually resulting from an imbalance in the normal rate of pleural fluid production or absorption, or both. Pleural effusions are associated with a number of pathological medical conditions. In the vast majority of these cases the cellular response is quite nonspecific and will show varying proportions of mesothelial cells, macrophages, erythrocytes, lymphocytes, neutrophils and other leukocytes. Certain diseases may occasionally show cellular changes in the effusion that will reflect their presence. Examples of these include infections such as tuberculosis, blastomycosis, aspergillosis, viral infections, echinococcosis and cryptococcosis. Noninfectious, nonneoplastic diseases diagnosable by cytopathologic methods include rheumatoid arthritis, luAPSTRAKTI 89 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. pus erythematosus, eosinophilic pleural effusion, pleural endometriois and sickle cell disease. In spite of these occasional specific cytopathologic diagnoses which can be rendered for the nonneoplastic diseases named above, the most significant role of cytopathology by far is in the examination of effusion fluid for the presence of cancer cells. Cytologic examination of pleural effusions is an important initial step in management of malignant effusions. Although most patients with a malignant effusion have a known history of cancer, a positive effusion may be the first sign of an unsuspected malignancy. Cytologic examination of a serous effusion may offer the possibility of an early and accurate diagnosis by using a minimal intervention. The presence of pleural effusions typically signals an advanced stage of disease so it is associated with poor prognosis. The accuracy of cytologic examinations of malignant pleural effusion is in high range (from 40% to 87%). Lung, breast, ovarian, and gastrointestinal cancers are most likely to cause malignant effusions. The histologic type of cancer most commonly seen in serous effusions is adenocarcinoma but a variety of other cancers can cause effusions. Less common malignancies are squamous cell carcinoma, small cell carcinoma (SCC), hematopoietic malignancies, melanoma, germ cell tumours and sarcomas. Mesotheliomas often present with recurrent serous effusions. One of the most important diagnostic doubts in pleural effusion analysis is distinguishing adenocarcinoma from mesothelioma and mesothelioma from benign mesothelial cells. In most cases the diagnose is based on regular cytomorphology analysis and immunocytochemistry is necessary in low percent of cases to establish the diagnose. The specimens are collecting by thoracentesis, processing in a routine fashion and staining by hematoxylin-eosin (H&E) and Papanicolaou stain after wet fixation with 95% ethanol and May-Grunwald-Giemsa stain after air drying. It is important to evaluate the same cells when constructing coordinate immunoreactivity pattern for various immunomarkers. Since the same cells cannot be followed on different cytology smears, they are not ideally suited for routine immunocytochemical evaluation of effusion fluids. Serial sections of cell-blocks show the same cells in adjacent levels which allow proper evaluation of the coordinate immunoreactivity pattern. Conclusion: Pleural fluid cytological analysis has important diagnostic role in most cases so primary aim should be to establish the correct diagnosis with minimal investigation. Cytology is the only method by which one may retrieve cells from fluid, often in the absence of solid lessions or when obtaining a biopsy from the latter is more difficult and costly and entails much greater discomfort or risk for the patient. Effusion cytology is important for: primary diagnosis of cancer, staging, diagnosis of recurrence or disease progression. Evaluation of effusion cytology is one of the most challenging areas in diagnostic cytopathology. A remarkably wide cytomorphological spectrum of reactive mesothelial cells overlaps with various benign and malignant processes. Due to these limitations, a significant proportion of effusion fluids are difficult to interpret with objective certainty by cytomorphology alone. The use of immunocytochemical panel of antibodies with combination of high sensitivity and specificy is the only way for precise distinguishing of adenocarcinomas from mesothelioma, especially in cases with scant tumor cells particularly in metastatic disease from well differentiated carcinoma, and reactive mesothelial cells from mesothelioma. Other stains are used to confirm pulmonary origin and should be selected as a complement to the panel once adenocarcinoma is confirmed. Molecular diagnostics are feasable on routine cytopreparations, primary diagnostics as well as molecular monitoring during targeted therapy.

The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC), which attempts to standardize reporting and cytological criteria for fine-needle aspiration of thyroid nodules and was first introduced in 2009, has been updated. Although much of the original TBSRTC remains the same, several “enhancements” have been introduced in the 2017 version based on new data and developments in the field. The 2017 revision reaffirms that every thyroid FNA report should begin with one of six diagnostic categories, the names of which remain unchanged since they were first introduced: nondiagnostic or unsatisfactory; benign; atypia of undetermined significance (AUS) or follicular lesion of undetermined significance (FLUS); follicular neoplasm or suspicious for a follicular neoplasm; suspicious for malignancy; and malignant. In the frst edition of TBSRTC, the implied risk of malignancy (MOP) for each diagnostic category was calculated and provided as a range based on a review of the literature at that time: 0–3% for benign, ~5–15% for atypia of undetermined signifcance (AUS) or follicular lesion of undetermined signifcance (FLUS), 15–30% for follicular neoplasm or suspicious for follicular neoplasm, 60–75% for suspicious for malignancy, and 97–99% for the malignant category. In the second edition, these ranges have been revised, especially for the so-called “indeterminate” categories, representing estimates calculated primarily from studies of large case cohorts and meta-analyses of ultrasound-guided thyroid FNA published after 2007. Notably, the new document reinterprets the previous version in one major way, and that is TBSRTC’s careful accommodation of the new noncancer category of noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) tumors, which prior to April 2016 were categorized as thyroid cancer. NIFTP tumors have nuclear changes on cytologic evaluation that are identical to other forms of thyroid cancer, but on close long-term clinical follow-up they do not appear to recur or metastasize, and therefore, they do not behave clinically like thyroid cancer. NIFTP tumors typically fall into categories 3, 4, or 5, and can only be diagnosed as “not cancer” after a full surgical excision is performed and the entire tumor specimen is examined under a microscope. There have also been a number of other enhancements with the 2017 update: • The option of molecular testing in the standard management of AUS/FLUS and FN/SFN has been included. • The definition and diagnostic criteria for FN/SFN has been modified: cases demonstrating mild nuclear changes associated with papillary thyroid carcinoma are now included. The definition and diagnostic criteria for the papillary thyroid carcinoma subset of the malignant category now suggest limiting use to cases with “classical” features of papillary thyroid carcinoma. TBSRTC is now the most common classification worldwide for the reporting of thyroid FNA specimens. In view of this, ABSTRACTS 92 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. new suggestions will be useful for various aspects of thyroid FNA including nomenclature, differential diagnosis, the potential impact of NIFTP on the indeterminate diagnostic categories, utility of molecular and IHC markers, and clinical management.

Breast cancer (BC) is the most prevalent cancer in the world among women and there are nearly 1.7 million new cases worldwide each year. Due to a number of remarkable advances made in both diagnosis and therapy, the survival rates for BC patients have increased in those regions with adequate medical facilities. According to contemporary recommendations, any pathological diagnosis of breast lesions, before any treatment, should be based on a Core Needle Biopsy (CNB), or on a Fine Needle Aspiration Cytology (FNAC), if CNB is not available. The prognosis of the newly diagnosed breast cancer patient depends on a number of factors, among which the most important is the extent of the spread of the disease to the axillary lymph nodes. Because any further treatment is influenced by the presence and number of axillary lymph nodes involved, a complete evaluation of the axillary lymph nodes is performed on every patient that is able to tolerate it, after a formal diagnosis of invasive carcinoma. At the very least an ultrasound with guided fine needle aspiration or core biopsy of suspicious lymph nodes should be undertaken.Although CNB is the main method employed in breast lesions diagnostics, FNAC still plays a significant role in the evaluation of pathological processes in the breast, a fact that has been well documented in the relevant literature in the last 20 years. The advantages of FNAC are: the sampling is quicker; the sampling technique usually does not require the use of anaesthetics; the trauma is small, and therefore more convenient for women using anticoagulant therapy; complications are rare; the availability of the results is within a few hours; skilled operators and pathologists regard this method as being highly sensitive in the detection of any malignant cells and the equipment is less expensive. The United Kingdom National Health Service Breast Screening Program (UK NHSBSP), began in 1988. Its guidelines have been published with regards to the mode of categorizing cell changes that may be seen in cytological samples obtained by needle aspiration. Five categories have been identified: C1 (unsatisfactory specimen - non-representative), C2 (benign), C3 (atypical - most likely benign), C4 (suspected - most likely malignant) and C5 (malignant). In 1996, the American National Cancer Institute (NCI) also suggested 5 categories for cytological diagnostics of breast lesions: benign, atypical, suspected, malignant and unsatisfactory. Patients with C3 and C4 categories, namely, atypical and suspected, which carry the risk of a malignant tumour, need to undergo further examination. C1 and C2 categories have to be correlated with the results of clinical and radiological examinations. C3 and C4 categories should not be represented in more than 5% of all analyzed aspirates. Currently, there is no individual morphological criterion that cytological diagnostics of malignant breast tumours could be based on. The most important cytological criteria that indicate whether it is a benign pathological process or a malignant tumor are: cellularity of the sample (a very important criterion, but it should be carefully interpreted), loss of cell cohesiveness (characteristic of malignant tumors), cellular arrangements, cell size, biphasicity in smear, the characteristics of the nucleus (size, contour, the appearance of chromatin, the appearance of nucleolus), characteristics of cytoplasm, nuclear-cytoplasmic ratio, APSTRAKTI 93 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. mitotic figures, background appearance (necrosis, peripheral blood cells, mucus…) and the presence of inflammatory cells. It is also possible to perform immuno-histochemical staining on cytological samples, flow cytometry and molecular analyses. The FNAC treatment is characterised by solid sensitivity, specificity and predictive value. The sensitivity of FNAC ranges from 89% to 98% and the specificity is between 98% and 100%. Major shortcomings of this method are the impossibility of diagnosing in situ carcinoma and lesions followed by any abundant production of connective tissue. The CNB treatment has gained remarkable popularity since the 1980s and in many institutions has replaced FNAC. The limitations of both methods are; atypical ductal hyperplasia, fibroepithelial tumours, radial scarring and papillary lesions. In the diagnosis of breast lesions apart from aspiration cytology, other sampling techniques for cytological analysis are also applied. In the era of breast conservation therapy, breast tissue is most commonly sent for intraoperative consultation. A frozen section analysis is performed through freezing and sectioning the surgical specimen with subsequent staining, in order to obtain an extemporaneous assessment of the margins. Although this technique is extensively used by many surgeons to avoid the need for a postponed rescission, some pitfalls have been reported, such as the occurrence of artefacts due to the freezing and thawing of the adipose tissue in the specimen. A different intraoperative method for margins evaluation is imprint cytology, which consists of pressing each of the 6 faces of the specimen on 6 different slides, so that any malignant cell on any involved margin is theoretically present on the cytology of the respective slide, because of the tendency of tumour cells to adhere to glass as compared to adipocytes. Imprint cytology can also be used in assessing the representational value of the CNB samples. A significant number of authors suggest that the application of the imprint of cytology reduces the number of inadequate samples obtained by CNB and can also provide a preliminary diagnosis, especially in cases of adequately sampled malignant tumours. Nipple discharge (ND) accounts for approximately 5% of the breast-related symptoms and is the third most common reason women seek medical attention. Approximately 7% to 15% of unilateral NDs are caused by malignant lesions, primarily ductal carcinoma in-situ (DCIS). A cytological examination of the obtained content is significant in the final treatment decision. Cytological analysis, in particular FNAC, continues to play an important role in the diagnoses of breast cancer. Skilled professionals can determine breast cancer through an analysis of the cytological sample as a reliable and accurate method.

Aim and introduction: Pediatric nodal marginal zone lymphoma (NMZL) is a rare, but distinct subtype of NMZL with characteristic clinical presentation, pathohistological and molecular features, therapy and prognosis. Results: We report the case of a 15-year-old boy with no remarkable past history, presented with painless enlargement of left submandibular lymph node (LN) for three months. He was admitted to the University Children’s Hospital in Belgrade in May 2016. The cervical ultrasound demonstrated moderate left submandibular lymphadenopathy, but also mild enlargement of two right submandibular LNs (17x7mm, 14x7mm). Physical examination, chest radiography and abdominal ultrasound revealed no hepatosplenomegaly and lymphadenopathy elsewhere. The result of blood count test was normal. Biochemistry showed elevated uric acid 499 umol/l, AST 45U/l, ALT 98U/l, and sedimentation rate (65mm/h). Urea, creatinine, alkaline phosphatase, LDH and CRP were normal. The patient underwent left submandibular LN excisional biopsy. The size of the LN was 47x37x20mm. The histopathological examination revealed partial architectural effacement: follicular hyperplasia and nodular B-cell infiltration with features of progressive transformation of germinal centers (PTGC) in the form of fragmentation of follicles. A CD20 immunostain shows an abnormal expansion of the marginal zone with infiltration of interfollicular space. These B-cells were negative for CD3, CD5, CD23, EBV-LMP1, bcl-6, CD10, EMA, CD30, CD15, MUM-1, and positive for bcl-2 and IgD. A CD21/ CD23/ fascin immunostain showed an expanded and disrupted follicular dendritic cell meshwork. Ki-67 highlighted residual follicular polarisation and a low proliferation rate in the interfollicular areas. Based on these pathohistological findings it was concluded that LN likely represent reactive follicular hyperplasia with atypical marginal zone hyperplasia or possible PNMZL, with APSTRAKTI 95 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. recommendation of polymerase chain reaction (PCR) analysis of clonality. Additional IGH PCR analysis demonstrated biclonal heavy chain gene rearrangement. These findings were consistent with PNMZL. After consultation with members of International BFM study group for non-Hodgkin lymphomas, followup was recommended without any treatment. The patient has remained disease free for 22 months since diagnosis. Conclusion: We presented a rare case of PNMZL with morphological features of PTGC, but immunohistochemistry and additional PCR clonality analysis were crucial for final diagnosis. This case represents a diagnostic and therapeutic challenge because of their rarity in the pediatric population.

Adnexal tumors of the skin comprise heterogenous group with over 40 defined entities, classified by predominant differentiation into lesions with apocrine and eccrine, follicular, sebaceous, or multilineage differentiation. Some, but not all these entities are represented by benign and malignant counterparts. Their occurrence may be sporadic or as a part of inherited syndromes (e.g. Muir-Torre syndrome, Brooke-Spiegler Syndrome, or Cowden’s syndrome). Adnexal tumors may arise de novo or within hamartomatous lesions such as nevus sebaceous of Jadassohn. Adnexal carcinomas are very rare tumors (the incidence is less than 0.001%), with variable local recurrence, metastatic potential, and survival. Porocarcinoma, hidradenocarcinoma and sebaceous carcinoma (especially ocular type) are considered to have a poor prognosis, with the highest risk of local recurrence and distant metastases. Mortality of the patients with porocarcinoma is very high (65-80%) if regional or distant metastases are present. The treatment of malignant adnexal tumors is usually surgical or less frequently with radiation therapy. Patients with metastases are usually treated with chemotherapy, mostly with cytotoxic reagents, and rarely with estrogen receptor antagonists. The detailed knowledge of genetic features of adnexal tumors is still lacking. Most of the studies examined only few of the genes using low throughput techniques. Development of new generations of genome sequencing methods led to better understanding of tumors with apocrine and eccrine differentiation. For many of their subtypes, it is still unknown whether there are specific genetic changes, that could even be of diagnostic significance. Hotspot mutations in HRAS (p.G13X and p.Q61X) were found in a subset of eccrine poromas and porocarcinomas. These mutations were found in tumors with other lines of differentiation and suggesting overlapping genetical characteristics among adnexal tumors. Due to their similar histological features, cylindroma and spiradenoma are usually considered as phenotypic variations of the same entity. Their histological features can be mixed, in which case a diagnosis of spiradenocylindroma is made. In cylindroma, MYB is upregulated either by mutations in CYLD gene (syndromic cases) or due to a rearrangement of MYB gene (sporadic cases). Genetic characteristics of spiradenomas, including the status of CYLD and MYB genes, are largely unknown. It is still unclear if these two are both histological and genetical “relatives” and what is the level of heterogeneity among tumors arising sporadically or within syndromes. The presence of chromosomal rearrangements in adnexal tumors is also unexplored. TORC1-MAML2 and EWSR1-POU5F1 fusions were found in significant number of hidradenomas. Initially it was thought these fusions could be characteristic for clear cell variant of hidradenomas, but no true correlation with histology was found. Molecular alterations that differ between benign and malignant counterparts and could enable targeted therapy of adnexal carcinomas are unknown. Mutations in TP53, often UV-associated, are frequent in malignant tumors with eccrine and apocrine differentiation and can drive malignant transformation in such tumors. Porocarcinomas and ABSTRACTS 96 MATERIA MEDICA • Vol. 34 • Issue 1, suplement 1 • april 2018. hidradenocarcinomas harbor various molecular alterations affecting PI3K-AKT or MAPK pathways that could enable targeted therapy in the future. Actionable mutations in EGFR were not found in carcinomas with eccrine and apocrine differentiation thus far. Her2 amplifications are rarely found, mostly in hidradenocarcinomas, but its therapeutic potential has only been utilized only once.16,25 Mutations of PTCH1 and TCF7L1 in hidradenocarcinomas could also enable the treatment with the inhibitors of Hedgehog and WNT/Hippo signaling pathways. It seems that current knowledge gained from genomic studies of adnexal tumors is only a scratch on the surface. In addition, there is no data on epigenetic characteristics or transcriptome of adnexal skin tumors. Taken altogether, further and detailed investigation of genome, epigenome and transcriptome of adnexal tumors is necessary. Such integrated knowledge could explain mechanisms of their development, malignant alteration and progression, so the treatment of patients with metastatic adnexal carcinomas could be changed toward targeted therapy.

Lung transplantation remains the definitive treatment for end-stage lung diseases and an option when
medical and surgical care has been exhausted. The first human single lung transplant was performed in
1963, and the patient, survived for 18 days. From 1963 to 1978, multiple attempts at lung transplantation
failed because of rejection and problems with anastomotic bronchial healing. It was only after the invention of the heart-lung machine, coupled with the development of immunosuppressive drugs, that organs
such as the lungs could be transplanted with a reasonable chance of patient recovery. The first clinically
successful long-term single lung transplant was performed in 1983, and since then over 25,000 lung transplants performed worldwide.

The presentation will reflect on a one-month period of education that the author spent with the Cleveland Clinic soft tissue pathology team. Cleveland is a US city in the state of Ohio. One of its nicknames is
“The Rock and Roll Capital of the world”, due to the fact that the term R’n’R was coined in the 1950s by
a Cleveland-based disc jockey Alan Freed. The city hosts the Rock and Roll Hall of Fame, established in
1983. It is also home to the Cleveland Clinic, a multispecialty academic hospital currently ranked as the
#2 hospital by U.S. News & World Report1. In 2014, Cleveland Clinic had a total revenue of $11.63 billion, making it the #2 hospital in US on the Becker’s Hospital Review revenue list2. The author spent one
month on a UICC ICRETT fellowship in November 2016 with the Cleveland Clinic soft tissue pathology
team. The main strength of the soft tissue team is the presence of several internationally known experts
with diverse interests within the field of soft tissue and beyond, with team philosophy highlighting the
synergy of team work and individual reputation. Among various topics that were covered during the
one-month fellowship, certainly one of the most interesting was differentiation among different fibrohistiocytic neoplasms. Fibrohistiocytic tumors are among the most frequent soft tissue tumors and they
are most commonly encountered in the skin. “Fibrohistiocytic” is in fact a merely descriptive term for
cells that resemble both normal fibroblasts and histiocytes, and not a true line of differentiation3. Like
other soft tissue tumors, fibrohistiocytic neoplasms are divided into benign, intermediate and malignant
categories. In presentation, the author will reflect on the key points in the pathology diagnosis within this
category of tumors, and these are:
- being able to give a common denominator to numerous variants of benign fibrous histiocytoma
- awareness of the pitfalls in the diagnosis of dermatofibrosarcoma protuberans
- discrimination of malignant fibrohistiocytic skin-based tumors from other, more adverse cutaneous
malignancies.

The Norvegian University of Science and Technology is the largest educational institution in Norway. It was founded in 1760 as the Trondheim Academy. The Faculty of Medicine and Health Sciences is part of the St Olav’s Hospital in Trondheim, and being there, as participant of the Annual Cytology Tutorial of the European Federation of Cytology Societies, was an outstanding experience. Colleagues from all over the world had the opportunity to meet and learn from experts in various fields of cytology. Particularly, differences between conventional and Thin Prep Pap smears, as well as immunocytochemistry of air-dried smears were thoroughly discussed.

Aim: To review current terminology of mixed exocrine and endocrine tumors of the large intestine. Introduction: Previous classification of colorectal tumors contained category called “mixed adenoneuroendocrine carcinoma” (MANEC) which encompassed neoplasms of the large intestine with features of both adenocarcinoma and a neuroendocrine carcinoma. Indeed, the vast majority of the mixed colorectal tumors have these two malignant components. However, this designation is no more suitable as other combinations of neuroendocrine and non-neuroendocrine tumors are recognised. Material and Metods: A detailed review of the literature on classification of mixed neuroendocrine-nonneuroendocrine tumors has been done. Results: The nonneuroendocrine component in a mixed colorectal tumor can be either exocrine or squamous and can be either benign or malignant. The histological grade of the nonneuroendocrine component may also vary. Therefore in several recent papers a new term has been coined “mixed neuroendocrine-nonneuroendocrine neoplasms” (MiNENs) in order to convey all possible combinations of the two components. According to the histologically estimated malignant potential, MiNENs are further subdivided into three categories low grade, intermediate grade and high grade. Conclusion: The new terminology is much more comprehensible than the previous ones and ensures a more accurate assessment of biological behaviour of the mixed colorectal tumors thus avoiding overtreatment of clinically innocent lesions.