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.

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: 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 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.

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.

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. 

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.

The design and construction of new biological systems in the way engineers design electronic or mechanical systems is the primary goal of synthetic biology. The ability to create and modify life forms and easy access to information to do so has raised a number of issues related to ethics and security. In the era of rapid development of biotechnology, and the perception of the consequent risks to the environment and health, the ethics of knowledge becomes a matter of practical significance. The concern about the misuse of knowledge from synthetic biology influences new risk reduction strategies, which can have significant effects on scientific progress. This paper will provide an overview of the main bioethical and biosafety issues of synthetic biology.