Search

INTRODUCTION AND AIMS: Endemic nephropathy (EN) which was found in strict geographical areas in Balkan region is an environmental form of aristolochic acid nephropathy (AAN). Recently, we proved that the presence of aristolactam DNA adducts is the firm evidence for the exposure to aristolochic acid (AA), the confirmed etiological agent of EN i.e. AAN. Upper urothelial cancers (UUC) which occur in 50% of EN patients can be a solitary manifestation of exposure to AA. It has not been resolved why EN should be restricted only to several limited rural areas and it was questioned whether EN might unrecognized occur in villages out of established endemic focuses. Our aim was to confirm occurrence of this sporadic form of EN in villagers from Croatia and Bosnia analyzing the presence of aristolactam DNA adducts in kidney cortex of patients with UUC. METHODS: In this retrospective study (2005-2010), the samples of 72 UUC from 69 patients (27 men, 42 women) residing from Croatia and Bosnia were analyzed. The data on medical history, place of birth and residence as well as laboratory data were taken from hospital medical records. The histopathological data were reviewed by two pathologists. 46 patients had available specimens of renal cortex, and aristolactam (AL)-DNA adducts were quantified by (32)P- postlabeling and the adduct was confirmed by mass spectrometry. RESULTS: Out of 72 tumors, 37 and 35 cases were from EN and non-EN areas, respectively. Mean age of patients was 72 (41-84), no statistical difference was found between EN and non-EN patients (p=0.2398). Also no significant differences were found in tumor grade (EN (N): LG 9, HG 26 ; non-EN (N): LG 14, HG 18, v2=2.412 ; p=0.12) or tumor recurrence rate (EN vs. non-EN 16.2% vs.22.9%, v2=0.5064 ; p=0.47), as well as in patients’ kidney function (eGFR- MDRD) at the time of surgery (EN vs. non-EN 41.1 vs. 53.1 ml/min/1.73m2 ; p=0.2044). There were more women within the EN group (v2=3.9679 ; p=0.046) while hemoglobin values were higher in non-EN group (110 vs. 123 g/l ; p=0.0391). Positive AL-DNA adducts were found in renal cortex of 63% (29/ 46) patients with UUCs. As expected it was found in 18/28 patients from EN areas but importantly also in 11/18 patients from non- endemic villages. CONCLUSIONS: Based on the presence of aristolactam DNA adducts in 63% analyzed samples it can be concluded that aristolochic acid is the most frequent cause of UUC in this cohort of peasants from Croatia and Bosnia. Finding of aristolactam DNA adducts in patients residing out of established EN villages is the first molecular evidence for occurrence of EN i.e. AAN much broader than previously considered. This is an important alert for nephrologists, urologist and public health authorities which are taking care of villagers from all world regions where plants of Aristolochia genus are growing.

ntroduction and Aims: Upper urinary tract urothelial cancers(UUC) are rare in general population but occur in almost 50% of patients with endemic nephropathy(EN)/aristolochic acid nephropathy (AAN). Aristolochic acid(AA) was proven as the etiological factor both for EN/AAN and UUC. The goal of this study was to identify tumor suppressor genes(TSG) and oncogenes(OG) involved in AA induced carcinogenesis and their posttranscriptional regulation mehanisms by microRNAs. Methods: Paired samples of tumors and adjacent normal urothelial tissues of 13 patients from Croatian and Bosnian endemic regions were analyzed. MiRNA profiling was performed by high- capacity quantitative PCR, while mRNA expression profiling of the same RNAs was done using microarray technology. Immunohistochemistry was performed on tissue microarrays. Results: The broad involvement of known cancer genes in the process of UUC tumorigenesis was revealed by expression profiling analysis of the tumors, implicating on many OG(35 up-regulated ; 54 down- modulated), and TSG(17 up-regulated ; 3 down- modulated). We studied whether the upregulation of specific OG (MYC, FGFR3, HRAS and KRAS) and down regulation of TSG (PTEN and PTCH1) might be augmented in UUC by coordinated action of miRNAs. Figure 1. shows a correlative network implicating numerous down-modulated miRNAregulators of these OG, including tumor suppressor miRNAs miR-23b, miR-143 and miR-145, and also a broad inhibition of PTCH1 by increased oncogenic miRNAs including miR-21, miR-18a and miR-9, and of PTEN by oncogenic miRNAs of the miR- 17-92 family. Both PTEN and PTCH1 appear commonly inhibited by essentially all the members of the miR-200 family. The elevated MYC then represents a central node in these processes, functioning as the activator of miR-17, miR-20 and miR-9 and inhibitor of miR-23b, the repressor of FGFR3 and KRAS, and the tumor- suppressor miRNA let-7c. By means of immunohistochemistry a trend of lower expression of PTEN protein was observed in tumor versus the normal tissue. Conclusions: AAN-associated UTUC carcinogenesis is characterized by a major deregulation of oncogenes and tumor suppressor genes, whose activity may be broadly and modulated by coordinate action of miRNAs.

Dietary intake of Aristolochia plants containing aristolochic acid (AA, IARC Group 1 carcinogen), leads to renal disease and to recurrent urinary tract transitional cell carcinomas (TCC). Mutation screens identified frequent A>T transversions, alterations underrepresented in other cancer types, as the signature of AA exposure. AA- associated TCC is a preventable cancer yet millions are estimated at risk due to unregulated medicinal use of Aristolochia. Here we present an ultra-low coverage modification of whole-exome sequencing (ULC-WES) as a novel molecular epidemiology method that reliably detects the AA mutation signature in small amounts of DNA from FFPE tumor tissues and can identify new exposed populations. DNA was isolated from paraffin sections of 18 TCC tumors from endemic nephropathy (EN) patients from regions in Croatia and Bosnia linked to exposure to AA. Four non-EN upper tract TCCs were used as controls. 250 ng of DNA was used for sequencing libraries and exome enrichment (Nimblegen, Roche). Libraries were sequenced on Illumina HiSeq2500 at 10-12x coverage. After data processing, GATK was used to call single nucleotide variants (SNV). To assess platform independence, 7 samples were run on SOLID 5500XL (Life), using proprietary chemistry and pipeline for data processing and variant calling. Normal genetic variants were filtered out after SNV annotation. ULC-WES at 10x identified the A>T predominance in 17 of 18 EN tumors (94%), and at T pattern in 14 (78%) samples. In contrast, the A>T presence was T mutations in the genome (R^2=0.83 for mutation count correlated with the chromosome length) but also identified recurrent non-synonymous A>T mutations in tumor suppressors and oncogenes including TP53, ARID1A, BRCA1/2, AXIN1, GATA3 and NRAS, DNMT1, FLT3, CSF1R and PIK3CA. We next detected recurrent A>T mutations in a number of histone and DNA modifiers suggesting epigenetic deregulation of genome activity in the AA-associated TCCs. In 2 patients, pairs of distinct TCCs from the same surgery had markedly overlapping mutation patterns (624 SNVs shared between same-side pelvic and ureteric tumors in one case, and 191 shared between renal pelvic and bladder tumors in the other). Thus, ULC-WES provided evidence supporting the concept of tumor cell seeding along the urinary tract as a mechanism of TCC spread and recurrence. We conclude that ULC-WES is a sensitive and specific method to identify AA exposure in tumor tissue and is directly applicable to identification of new populations at risk. The method can ultimately facilitate evidence-based preventive measures in the affected regions.

Endemic nephropathy is a syndrome that comprises two entities: chronic interstitial nephropathy and urothelial cell cancers predominantly of the upper urinary tract. The etiological agent for the disease is aristolochic acid, a compound found in the plants of Aristolochia spp. The development of urothelial cancers is characterized by the formation of aristolactam DNA adducts leading to mutations, predominantly A: T->T: A transversions. In order to comprehensively understand the gene regulation programs in upper urothelial cancers we performed integrated miRNA and mRNA expression profiling of paired tumours and unaffected urothelium samples. The obtained data will help us to understand the carcinogenesis caused by aristolochic acid and might be the source for the design of a diagnostic biomarker.

Background: Dietary intake of nephrotoxic and carcinogenic aristolochic acid (AA) leads to endemic nephropathy (EN) marked by chronic tubulointerstitial nephropathy (CTN) and urinary tract transitional cell carcinomas (TCC). The role of AA in malignancies other than TCC is unexplored. We aimed to investigate a role of AA in the etiology of renal cell carcinomas (RCC), usually not related to EN, by conducting whole exome sequencing (WES) of tumor-enriched DNAs to detect the presence of the AA mutational signature. Methods: Four clear cell RCC patients from EN area linked to AA exposure and 1 non-EN RCC case were studied. Of the EN patients, 3 were farmers baking bread from grain from locally grown wheat ; 1 patient was diagnosed with CTN and 2 with TCC. All patients were in CKD stages≥3b.Tumor DNA was isolated by macrodissection from FFPE sections, processed for WES libraries, exome capture and sequencing on Illumina HiSeq2500 (multiplexed paired-end 50 bp run, >10x coverage/sample). Reads were aligned by BWA, variants called by GATK, annotated by ANNOVAR and genetic variants observed in normal population removed. The detection of the AA signature was performed by customized R functions. Results: High A:T>T:A transversion rates (0.9 to 1.8 /Mb) were found in all EN RCC samples. Three cases had A:T>T:A (48.1%, 38.2% and 21.6%) and 1 tumor was borderline positive (15.2% A:T>T:A). The non-EN RCC control was negative (6.4% A:T>T:A). In the positive samples the mutations occurred mainly in the CAG context and on the coding strand, meeting all definition criteria for the AA mutational signature. Conclusions: Our study finds AA mutational signature in 3 EN patients with RCC, suggesting a possible causal involvement of AA in the RCC etiology, with new implications for the worldwide RCC incidence due to widespread unregulated use of AA-containing herbs. Extension of this study is underway to validate the preliminary results in a larger cohort of cases. Supported by grant 04-38 Cro Nat Foundation

The aim of this study was to correlate the expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor- 1α (HIF-1α) with pathological renal artery changes in patients with renal cell carcinoma (RCC). Moreover, we correlated intratumoral microvessel density (MVD) with VEGF and HIF-1α expression and prognostic factors for RCC, including tumor necrosis. Formalin-fixed and paraffin-embedded tissue blocks from 150 patients with RCC and 50 patients with nontumorous kidney diseases were analyzed. The control group consisted of specimens from both renal arteries obtained from 25 decedents at routine autopsy (50 cases in total). Immunohistochemistry was performed using primary antibodies to VEGF, HIF-1α and CD31. Results: Pathological renal artery changes are found more common in patients with RCC and nontumorous kidney diseases when compared to control group. MVD was higher in the RCCs of patients with pathological renal artery changes. Tumors with higher HIF-1α expression had higher MVD. However, the VEGF expression was not associated with MVD. Significant association was also found between MVD and the extent of tumor necrosis--less necrotic tumors had higher MVD. Association between renal artery changes and VEGF and HIF-1α expression was not established. Considering the results of this study the evaluation of renal artery changes in forthcoming research of RCC would be appreciated for several reasons: to estimate their incidence in a larger number of patients, to clarify their connection with RCC and to reveal their relationship with MVD in RCCs.

Objectives:To describe the clinical, radiologic, cytologic and histologic features of a case of mammary pseudoangiomatous stromal hyperplasia (PASH). Materials and Methods:The authors reviewed the clinical, radiologic, cytologic and histologic results in a case of PASH in 31-year old, nonpregnant woman presented with a painless nodule in left breast.The findings were compared with those in the literature. Results: Clinicaly PASH is indistinguishable from a fibroadenoma. Sonographic findings included ovoid, cirscumscribed, hipoechoic solid mass which is similar to fibroadenoma. Fine needle aspiration cytology was performed under sonographic guidance and the cytologic features were consistent with atypical fibroadenoma. The mass was surgicaly excised. Histologic analyses was performed and a diagnosis of PASH was made. Conclusion: A definitive diagnosis of a PASH is unlikely on the basis of the cytologic examination alone. To confirm the diagnosis of a PASH a biopsy is required. PASH needs to be distinguished from low-grade angiosarcoma and some borderline lesions which often have a similar cytologic appearance. Management of PASH is similar as for fibroadenoma – periodic mammographic and sonographic surveillance or surgical excision.

Endemic (Balkan) nephropathy (EN), a chronic renal disease affecting residents of rural villages situated near tributaries of Danube River, is strongly associated with transitional cell (urothelial) carcinoma of the upper urinary tract (UUC). Aristolochic acid (AA), a powerful nephrotoxin and human carcinogen, was shown recently to be the causative agent in EN. In EN, exposure occurs through ingestion of bread prepared from flour contaminated with AA. After metabolic activation AA forms covalent DNA adducts in renal cortex and urothelial tissues. Aristolactam-DNA adducts generate unique mutational spectra in p53 tumor suppressor gene, which together with the presence of DNA adducts in the renal cortex serve as biomarkers for aristolochic acid nephropathy and associated urothelial carcinomas. TP53 mutation spectrum was dominated by A:T→T:A transversions located almost exclusively on the non-transcribed DNA strand with unique „hot spots“ at several splice sites and at codons 131 and 209. TP53 gene mutations at this position have not previously been reported. The mechanism underlying the observed strand bias appears to be a selective failure to excise AL-DNA adducts by global genomic nucleotide excision repair. This factor also may account for the remarkable persistence of these adducts in human tissues (in some cases more than 50 years). In summary, aristolochic acid joins vinyl chloride and aflatoxin as human chemical carcinogens with a definitive mutational signature. This important information, coupled with the use of AL-DNA adducts as a biomarker, should prove useful in establishing the role of AA ingestion in countries with a high prevalence of UUC.

Introduction. Gastric polyps are uncommon, found in about 0, 4% of adult autopsies. The great majority are non-neoplastic and appear hyperplastic nature. Hyperplastic polyps form as a reaction to chronic inflammation in the cells that line the inside of the stomach. They are usually benign, but they can become cancerous if left untreated. Aim. The aim of this study in find out frequency of Helicobacter pylori infection in gastric polyps. Material and method. For cytology air dried, May-Grumvald Giemsa stained imprint of polyp biopsy were prepared. For histology parafin- embedded, 5um thick sections stained with hemalaun-eosin of polyp biopsy were prepared. Results. In period of 1st January 2000 to 31st December 2002 in our hospital 38 endoscopically removed gastric polyps were examined cytological by imprint method. All material were completely histologically examined. Polyps hyperplasticus was diagnosed in 37 histological slides, and there was one Adenoma tubular. In our speciment Helicobacter pylori was not found in 73, 7% gastric polyps, and was not found in 26, 3%. But, 70% polyps with Helicobacter pylori, i.e. 18, 4% of all polyps were situated in prepyloric region. Conclusion. In course of 3 years 38 gastric polyps were examined cytologically and histologically. Only 26, 3% of these patients had accompained Helicobactery pylori infection and 70% of these were situated at prepyloric region. So, we found that majority of gastric polyps ; mainly have not positive correlation with accompained Helicobacter pylori infection.

Endemic (Balkan) nephropathy (EN), a chronic renal disease affecting residents of rural villages situated near tributaries of Danube River, is strongly associated with transitional cell (urothelial) carcinoma of the upper urinary tract (UUC). Aristolochic acid (AA), a powerful nephrotoxin and human carcinogen, was shown recently to be the causative agent in EN. In EN, exposure occurs through ingestion of bread prepared from flour contaminated with AA. After metabolic activation AA forms covalent DNA adducts in renal cortex and urothelial tissues. Aristolactam-DNA adducts generate unique mutational spectra in p53 tumor suppressor gene, which together with the presence of DNA adducts in the renal cortex serve as biomarkers for aristolochic acid nephropathy and associated urothelial carcinomas. TP53 mutation spectrum was dominated by A:T→T:A transversions located almost exclusively on the non-transcribed DNA strand with unique „hot spots“ at several splice sites and at codons 131 and 209. TP53 gene mutations at this position have not previously been reported. The mechanism underlying the observed strand bias appears to be a selective failure to excise AL-DNA adducts by global genomic nucleotide excision repair. This factor also may account for the remarkable persistence of these adducts in human tissues (in some cases more than 50 years). In summary, aristolochic acid joins vinyl chloride and aflatoxin as human chemical carcinogens with a definitive mutational signature. This important information, coupled with the use of AL-DNA adducts as a biomarker, should prove useful in establishing the role of AA ingestion in countries with a high prevalence of UUC.

Publications report on the Bosnian endemic focus as more active than the Croatian during the last decades. Our aim was to assess the current state and to compare upper urothelial cancer (UUC) patients’ characteristics between Bosnian and Croatian foci. Design and Methods: Study included 40 patients undergone surgery for UUC in the period from 2005 to 2010 in General Hospital „Josip Benčević“ in Slavonski Brod. The definition of UUC comprises calices, pyelon and/or ureter cancer. Aristolactam (AL-I)-DNA adducts were detected and quantified in renal cortex using 32P-postlabeling method and additionaly confirmed by mass spectrometry. Modified MDRD (Modification of Diet in Renal Disease) formula was used to calculate glomerular filtration rate, and patients were classified in chronic kidney disease (CKD) stages groups according to the KDOQI (Kidney Disease Outcomes Quality Initiative) guidelines. Statistical analysis was performed using SPSS 16.0 at significance level p

Objectives and study: Approximately 50% of endemic nephropathy (EN) patients develop upper urothelial tract cancers (UUC) at some point during the course of their disease. Aristolochic acid (AA) was proven as the etiological factor both for EN and UUC. The goal of his study was to establish patterns of gene expression and posttranscriptional gene regulation mechanisms involved in AA induced carcinogenesis. Methods: Paired samples of tumors and adjacent normal urothelial tissues of 10 patients from Croatian endemic region were analyzed. miRNA profiling was performed by high-capacity qPCR using Applied Biosystems megaplex RT primer pools for 754 human miRNAs and the microfluidisc TagMan Low Density Arrays, while miRNA profiling of the same RNAs was performed using Affymetrix HG U133 Plus 2.0 arrays. Results: A signature of 50 miRNAs differentially modulated (19 elevated and 31 reduced) and 4801 significantly modulated mRNAs (2005 elevated and 2796 reduced) were identified in tumor vs. unaffected tissue. Using R-scripting, miRNA and mRNA dana were integrated, identifying 643 predicted, inversely correlated targets oft he 19 upregulated miRNAs and 483 predicted, inversely correlated targets oft he 31 downregulated miRNAs. Cancer-related miRNAs were upregulated in UUC while their tumor suppressor targets (PDCD4, PTEN, TPM1, PTCH1) were reduced. Additionally, the anti- metastatic miR-200 family was elevated in tumors, concurrently with low levels of its targets, the ZEB1/2 repressors of E cadherin. Inhibition of invasive/migratory process was documanted by increased miRNAs targeting cellular movement, migration and invasion (FGF1, COLIA2, CTGF, IGF1, MMP2, PTEN, RHOB, THBSI and TGFBR2). Comprehensive biological interpretation and mining oft he integrated miRNA:mRNA dana set (Ingenuity Pathway Analysis) identified high-scoring pathways of renal necrosis, renal damage and failure, cancer regulated cell growth and proliferation, G2/M DNA damage checkpoint regulation and DNA repair, inflammatory disease and suppression of cellular movement and migration. Conclusion: UUC tumorigenesis appears to involve primarily miRNA- mediated process of repression of tumor suppressor genes, tumor growth, induction of inflammatory processes and repression of anti- invasion/metastasis programs rendering the non- metastatic nature of these carcinomas.

Endemic (Balkan) nephropathy (EN) is characterized by an insidious onset, slow progression to end stage renal disease and a strong association with upper urinary tract cancers (UUC). EN occurs only in rural villages, situated near tributaries of the Danube River ; epidemiologic studies reveal a familial aggregation, but not an inherited disease. Various environmental agents, including ocratoxin A have been proposed as putative causative factors of EN but strong evidence in support of these hypotheses in lacking. We have proposed that chronic dietary exposure to aristolochic acid (AA) is genetically susceptible individuals is uniquely responsible for EN and UUC. This hypothesis is supported by the unquivocal detection of 7-(deoxyadenosin_N6-yl) aristolactam DNA adducts in renal cortical and UUC tissues of patients with EN and/or UUC.

Hyperparathyroidism is a syndrome caused by adenoma, hyperplasia or rarely by cancer of the parathyroid glands. Secondary hyperparathyroidism is a common complications in cases of chronic renal insufficiency, with clinical and biochemical signs of increased function of parathyroid glands. It may be so pronounced that surgery is necessary. Sometimes medical treatment of hyperparathyroidism consists of percussing inactivity of tumor of the parathyroid glands with alcohol. Both procedures demand preoperative location of the tissue of the parathyroid glands. Preoperative location of abnormal, nonpalpable parathyroid glands in patient with hyperparathyroidism is largely dependent on use of a high-resolution sonography. However, such a technique has some limitations and aid of a fine-needle biopsy (FNAB) may be relevant.

The activation of protooncogenes and inactivation of tumor suppressor genes are considered to be the main molecular events in the multistep process of carcinogenesis. Mutations of the TP53 tumor suppressor gene have been found in nearly all tumor types and are estimated to contribute to more than 50% of all cancers. Most mutations lead to the synthesis of highly stable, inactive proteins that accumulate in the nucleus of cancer cells. Alterations of codons 175, 248, 273 and 282 correspond to 19 % of all mutations and are considered general hot spot mutations. Dietary exposure to aristolochic acid (AA), an established nephrotoxin and human carcinogen found in all Aristolochia species was shown to be the causative agent of aristolochic acid nephropathy (previously called Chinese herbs nephropathy). This syndrome is characterized by proximal tubular damage, renal interstitial fibrosis, slow progression to the end stage renal disease and a high prevalence of upper urinary tract urothelial carcinoma (otherwise a highly unusual location). AA preferentially binds to purines in DNA and is associated with a high frequency of A → T transversions in the p53 gene. Rats treated with AA develop A:T → T:A mutations in codon 61 of c-H-ras gene. The pathological and clinical features of endemic (Balkan) nephropathy closely resemble those associated with aristolochic acid nephropathy except for the slower progression to end stage renal disease and longer cumulative period before the appearance of urothelial cancer. Recently, we reported the presence of AA-DNA adducts in renal cortex and A → T p53 mutations in tumor tissue of patients from Croatia and Bosnia with endemic nephropathy. These data support the hypothesis that dietary exposure to AA is a major risk factor for endemic (Balkan) nephropathy.

It was known for several decades that populations of endemic areas have significantly higher risk for upper urothelial cancers than non-endemic populations.

Introduction: Endemic nephropathy (EN) is associated with an increased frequency of urothelial carcinoma (UC) of upper urinary tract (UUT). Histologically, atrophy of proximal tubules and diffuse cortical fibrosis is the most prominent finding. Aim: Aim of the study was to analyze renal parenchyma of kidney specimens from patients with UC of the UUT from endemic region. Materials and Methods: We analyzed consecutive series of 17 cases (M:F=9:8) of UC of UUT from EN region who underwent nephrectomy in the period from June 2006 to December 2007. These cases were compared with 15 cases (M:F=5:10) of UC of UUT from non-endemic region in Zagreb. In the group of patients from endemic region, 10 (M:F=5:5) patients were from endemic region in Bosnia and Herzegovina, and 7 (M:F=5:2) patients were from endemic region of Slavonski Brod county. Specimens were routinely fixed, embedded in paraffin, cut and stained with haematoxylin and eosin and Mallory trichrome method, and examined by light microscopy. Results: In the group of patients from endemic region 8 patients had grade 2, and 9 had grade 3 of UUT carcinoma. There were 9 cases of UC of ureter and 8 cases of the UC of pyelon. Extensive tubular atrophy was found in 9 cases. In the group of patients from non-endemic region extensive tubular atrophy was found in 2 cases. Prominent interstitial fibrosis characterized 8 cases, compared to 2 cases from non-endemic region. Glomerular sclerosis was observed in 8 cases, compared to 2 cases in non-endemic region. We found signs of thyreoidiziation with prominent interstitial cell infiltration, mostly with mononuclear inflammatory cells in 7 cases compared to 3 cases in non-endemic region. Conclusion: Renal pathomorphology of patients with UUT carcinomas from endemic region differs from patients from non-endemic region. Extensive tubular atrophy and prominent interstitial fibrosis are characteristic findings in patients suffering of UUT carcinomas in endemic region. There were more cases of UC from endemic region from Bosnia and Herzegovina. Due to war and population migration, patients with UC and EN could become patients in other hospitals in Europe. Key words: endemic nephropathy, upper urinary tract carcinomas, patomorphology

Endemic nephropathy (EN) is a chronic tubulo-interstitial disease of unknown etiology, and it is associated with increased frequency of urothelial carcinoma of upper urinary system. Histologically, atrophy of proximal tubules and diffuse cortical fibrosis is the most prominent finding. Hyperplastic arteriopathy of small and middle-sized arteries is also reported as a constant finding in EN cases. Renal artery changes are observed in high frequency in patients with renal cell carcinoma, as well as in patients with urothelial carcinoma of renal pelvis from non-endemic region. Aim of this study was to analyze renal artery changes in patients with urothelial carcinoma of the upper urinary system and endemic nephropathy.

Malignant melanoma is very rare in the gastrointestinal system ; it comprises about 0.2- 3% of all melanomas and has extremely poor prognosis. Surgical treatment ranges from radical resection with lymphadenectomy to local excision alone. Primary melanoma is most frequently found in anorectal region of the gastrointestinal system. Aim of the study was to analyze frequency and sites of primary gastrointestinal melanoma in our bioptic material collected during a 10 year period. We used the computer database from Ljudevit Jurak University Department of Pathology and Surgical Department, Sestre milosrdnice University Hospital, Zagreb in the time period from 1996 to 2005. During the analyzed period there were 2831 patients (M:F=1748:1083) with diagnosed gastrointestinal tumors. Female patients were between 22-86 years old (mean 62, 0) while males were between 26-84 (mean 68, 0). In the examined time period malignant melanoma was diagnosed only in 16 (0, 6 %) patients (M:F=12:4). At the time of diagnosis male patients with melanoma were between 22-79 years old (mean 54, 0), while females were between 42– 70 (mean 59, 0). Out of all patients with gastrointestinal melanomas only 5 (31.3%) cases (M:F=3:2) were primary gastrointestinal, while 11 (68.7%) cases were metastases from skin melanoma. Two cases of primary melanomas in gastrointestinal system were originated from gallbladder (M:F=1:1), while 3 cases were diagnosed in the anorectal region (M:F=3:1). Female patients with primary gastrointestinal melanoma were 52 and 57 years old, while males were between 52 and 75 (mean 66, 0). Metastastatic melanomas usually involved small intestine (9/11 cases), while only one case was found in stomach and one case in colon. Although primary gastrointestinal melanomas are very rare, they are almost never suspected on clinical examination. In our bioptic material primary melanomas were found in anorectal region and gallbladder, while metastatic melanomas were usually found in the small intestine.

INTRODUCTION: Schwannoma is a benign, usually encapsulated nerve sheath tumor originating from Schwann cells. Typically, schwannomas affect the cutaneous nerves of the head and neck, upper and lower extremities and trunk. They predominantly occur in females between the second and fifth decade of life. Visceral schwannomas are very rare, and according to literature were described in the heart, kidney and lung. The retroperitoneal space is another localization of schwannomas. Except in cases of von Reclinghausen´s disease, the adrenal localization of schwannoma is particularly rare. CASE REPORT: We present a case of a 55 year-old female patient with previously diagnosed breast cancer treated with mastectomy followed by lymphadenectomy. Histologically, breast cancer was classified as ductal invasive breast cancer, histological grade 2, without lymph nodes metastases. Hormone receptors (estrogen and progesterone) were highly positive while HER2/neu was negative. Clinical examination did not find any signs of neurofibromatosis. Patient was treated with hormonal therapy. Three months after mastectomy ultrasound examination revealed a hormonally inactive tumor in the left adrenal gland that was suspected for a metastatic process. After adrenalectomy, macroscopic examination revealed a partially encapsulated tumor within the adrenal gland with yellowish solid cut surface, measuring 5x4x3 cm. Microscopically, the tumor was composed of solid sheets and bundles of uniform spindle shaped cells, along with various vascular and inflammatory elements embedded in an extensive extracellular matrix. No Verocay bodies were found. Immunohistochemicaly, tumor cells were positive for S-100 protein. Extensive histological analysis revealed no ganglion cells. DISCUSSION: Histological and immunohistochemical analysis of described adrenal gland tumor was consistent with the diagnosis of schwannoma. Adrenal tumors are frequently incidental and asymptomatic discoveries, and their therapy is a subject of controversial discussion. The combination of adrenal gland schwannoma and primary breast cancer is very rare, very probably incidental, but preoperatively difficult to distinguish from metastasis.

AIM OF THE STUDY: Gastrointestinal tract is the most commonly involved extranodal site for non- Hodgkin lymphoma that usually affects the stomach, less frequently small intestine and very rarely colon or esophagus. Aim of the study was to analyze frequency and types of gastrointestinal lymphoma in our bioptic material during the last 10 years. MATERIAL AND METHODS: We used the computer database (Thanatos) from Ljudevit Jurak University Department of Pathology for the time period from 1996-2005 for all patients who underwent gastrointestinal tract surgical procedures at the Department of Surgery, Sestre milosrdnice University Hospital, Zagreb. RESULTS: In the examined time period there were 2828 patients (M:F=1750:1078) presenting with gastrointestinal tumors. Esophagus was involved in 108 cases, stomach in 720 cases, and 2000 were in the small intestine and colon. Gastrointestinal lymphoma was diagnosed in only 18 (0, 6 %) cases (M:F=9:9). Male patients were between 50 and 73 years of age (mean 60, 0 years) ; while females were between 49-82 (mean 65, 0). The majority (16/18) of gastrointestinal tract lymphomas were found in the stomach (M:F=7:9), while only two cases were diagnosed in the small intestine. Both patients with small intestine lymphoma were male, 64 and 67 years old. Histologically all examined lymphomas were classified according to the World Health Organization (WHO) as diffuse large cell lymphoma, B phenotype. CONCLUSION: Gastrointestinal lymphoma was more commonly diagnosed in the sixth decade of life, with stomach being the predominant site of involvement. In patients having MALT lymphoma diagnosed on endoscopic biopsy, surgical operation was not performed. Therefore confirmation of MALT lymphomas was impossible. Lymphomas involving the gastrointestinal tract, although rare, should be considered in differential diagnosis when dealing with gastrointestinal tract malignancies.

Tumor necrosis in renal cell carcinoma (RCC) is found to be one of the independent prognostic factors, associated with decrease in microvessel density and an increase of VEGF protein expression, manly within the perinecrotic rim (1). Main renal artery may show different lesions such as arteriosclerosis or fibromuscular dysplasia (FMD) but connection between renal artery changes and other prognostic factors in RCC is still unknown (2, 3, 4, 5, 6). We analyzed a consecutive series of 55 patients (M:F=35:20) with RCC who underwent nephrectomy in the year 2003. Patients were aging from 35-79 years (mean 59.8) and the tumor size was 2.5-16 cm (mean 7.1). Tumor necrosis was found in 43 (78.2%) cases of RCC of which 32 had less than 50% tumor necrosis and 11 tumors contained more than 50% necrotic areas. Nuclear grade of RCC was determined according to Fuhrmann et al (7). Specimens were routinely fixed, embedded in paraffin, cut and stained with hematoxylin and eosin, Mallory trichrome method and orcein. Immunohistochemical analysis was performed using primary antibodies to VEGF (DAKO, Copenhagen, Denmark). Immunohistochemical reaction was assessed as: 0-no positive tumor cells ; 1-up to 10% positive tumor cells ; 2->10-50% positive tumor cells ; 3-more than 50% positive tumor cells. Renal arteries of 23 (41.8%) patients (M: F=18:5) showed no changes (Group I). These patients were in age from 38-78 years (mean 58.7) and tumors measured from 2.5-17 cm (mean 7.4). FMD (Group II) was found in 25 (45.5%) patients (M:F=12:13) ranging in age from 35-79 years (mean 59.1), and the tumor size was 3-16 cm (mean 8.2). Atherosclerotic changes of renal arteries (Group III) were observed in 7 (12.7%) patients (M:F=5:2), aged from 60-69 years (mean 62.0) with tumors from 2.5-14 cm (mean 6.4). VEGF expression was negative (0-no positive tumor cells) in 14 cases, 19 cases had less than 10% positive tumor cells, 13 cases had 10-50% positive tumor cells and 9 tumors expressed VEGF in more than 50% tumor cells. Statistical analysis did not show significant difference in age, tumor size or nuclear grade between the three groups regarding renal artery status. However, tumors without renal artery changes were more frequently without necrotic areas (7/12), in comparison with other groups, but those tumors also showed higher VEGF expression in tumors cells (only 2/23 cases without renal artery changes showed negative reaction for VEGF expression). Tumors with FMD usually contained necrotic areas (22/25 cases), but they also had the highest number of negative tumor cells expression for VEGF (9/14 tumors with negative VEGF expression were tumors with FMD). Renal artery changes are very common in patients with RCC (8, 9). Tumor necrosis was more common in patients with associated FMD and atherosclerotic changes of renal arteries. However, although it would be expected that tumors with more necrotic areas produce VEGF protein, the VEGF expression in this group was more frequently negative. The cause of renal artery changes and their relationship to the degree of tumor necrosis and other VEGF expression in tumor cells should be further analyzed.

AIM: Micropapillary carcinoma represents an uncommon variant of urothelial carcinoma associated with higher grade and advanced stage at time of diagnosis, as well as with poorer prognosis. Even though the morphology may be deceptive, this tumor type is considered to be a tumor with an aggressive behavior. Aim of this study was to review patients with micropapillary variant of urothelial carcinoma in our database and compare them with patients with “ classical” urinary bladder carcinoma. MATERIALS AND METHODS: We used the histopathological database (Thanatos) from the Ljudevit Jurak Department of Pathology for all urologic patients with diagnosed urinary bladder cancer, during the time period from January 1st, 2004 to March 31st, 2006. RESULTS: In the examined period there were 390 patients with urinary bladder cancer (M:F=298:92). Male patients were ageing from 26 to 91 years (mean 68.1) while females were ageing between 35 and 90 (mean 69.0). From the total number of patients with urinary bladder cancer there were only 10 (2.6%) cases with the micropapillary variant (M:F=8:2). In the group of patients with micropapillary carcinoma, at the time of diagnosis, males were ageing between 57-88 years (mean 70.9), while females were in age 76 and 81 years (mean 78.5). All micropapillary carcinomas were invasive at the time of diagnosis, and in 60% of all cases infiltration of muscular layer of urinary bladder was present. In 3 (30%) cases, after the diagnosis of micropapillary carcinoma, radical cystectomy was performed followed by lymphadenectomy, while only 7.9 % (30) cases with “ classic” urothelial carcinoma were treated with radical operation. Lymph nodes metastases were confirmed histologically in all 3 cases with micropapillary cancer and in 38.1% (8/21) cases with classical urothelial cancer. CONCLUSION: In our series micropapillary carcinoma was an aggressive tumor, often in advanced stage of disease, with a high incidence of lymph node metastases. Therefore, correct diagnosis is important to perform adequate, usually radical treatment.

INTRODUCTION: Spindle cell hemangioma is a vascular tumor first described as a new entity in 1986 by Weiss and Enzinger, as a rare type of haemangioma. The tumor occurs usually in young adults and affects the subcutis of distal extremities or arose in the small bones of the hands, feet and tibia. Location within the canalis spinalis is extremely rare. CASE REPORT: A 31 year-old male patient complained on weakness of both legs that started in May 2005. In September 2005 he noticed paresthesias in both legs that slowly progressed toward his stomach. He was admitted at our Department of neurosurgery in March 2006 due to progressive weakness of his legs. Upon admission the neurological examination revealed a spastic paraparesis (3/5), hyperactive reflexes and paresthesias distal from Th10 dermatome. MR of the thoracic spine showed an intraspinal, extradural mass lesion, measuring 5, 3 x 1, 2 cm at the Th1-Th3 level. Intraoperatively, C7-Th3 laminectomy was performed. On the right side of the spinal canal, a purple-grayish tumorous lesion was visualized. It was located extradurally compressing the dura to the left. The lesion was well demarked from the dura, and it was removed completely in a microneurosurgical manner. Histologically H&E slides revealed a lesion which was composed of thin-walled cavernous vessels lined by flattened endothelial cells and containing thrombi. There were cellular spindled stromal areas between the cavernous spaces that were assumed to be representing collapsed small vascular canals. Some of endothelial cells contained prominent intracytoplasmatic vacuoles. Postoperative recovery was uneventful. Paresthesias regressed almost immediately after surgery and the amelioration of leg weakness was noticed. MATERIAL AND METHODS: Immunohistochemical analysis of CD 34, Vimentin, SMA and Ki - 67 (DAKO, Copenhagen, Denmark) was performed in formalin fixed and paraffin embedded tumor tissues. As a visualization system labeled streptavidin biotin method (LSAB) was used on Dako Tech Mate automatic immunostainer using microwave streptavidin immunoperoxidase-MSIP protocol. DISCUSSION AND CONCLUSION: occasionally the spindle cell hemangioma is associated with Maffucci syndrome and was also seen in the Klippel-Trenaunay syndrome. The tumor arose in the vicinity of clearly abnormal vessels, supporting the idea that the spindle cell hemangioma is most likely a vascular malformation in which variation in blood flow gives rise to alternating areas of vascular expansion and collapse. Although, usually it was believed to be a tumor with limited metastatic potential and it is regarded as a benign tumor, about 60% of spindle cell hemangiomas recur and there is no evidence that these lesions have the ability to metastasize. This case is reported as a unique location of this rare type of hemangioma in the spinal canal.

Balkan endemic nephropathy (BEN) is a chronic tubulointerstitial nephropathy with an incompletely known etiology. The aim of this study was to compare the occurence of upper urinary tract carcinomas between Brodsko-posavska region and Zagreb in two six-year periods, 20 years apart.

AIM OF THE STUDY: According to the department’ s cancer registry data in the last eight years urological cancers represented 32.2% (3015 cases) of all male cancers, and 8.7% (594 cases) of all female cancers. The aim of this study is to determine the frequency and the pattern of urologic cancers diagnosed at our department in two eight-year periods a decade apart. MATERIALS AND METHODS: Histopathological data for the two periods were obtained from the department’ s computer-based carcinoma registry, former being a period of 1980-1987 and the latter of 1998-2005. Patients were assessed according to the sex and the localization of the tumor. RESULTS: 1459 urologic cancers were reported over a period of 1980-1987 (83.6 % in males, 16.4% in females), opposed to 3609 cases (83.5% in males, 16.5% in females) diagnosed in the period of 1998-2005. The organ-specific distribution in males, in the former period, was as follows: urinary bladder 62.5%, prostate 24.7%, kidney 7.5%, testis 3.9% and finally, renal pelvis and ureter with 1.4%. In the latter period prostate cancer (diagnosed in 42.6% of cases) switched places with urinary bladder carcinoma (diagnosed in 36.9% of cases), with the remaining distributions’ ranks staying the same. One must emphasize that the frequency of prostate cancer rises noticeably in the last six years. In females a predominance of bladder tumors was found in both periods (74.2% of all urinary cancers in the first and 59.4% in the second period), followed by kidney and renal pelvis and ureter malignancies. DISCUSSION AND CONCLUSION: The absolute number of all urinary carcinomas increased three to four times during the observed periods, with the absolute number of prostate cancer quadrupling, mostly due to more sophisticated diagnostic methods. The bladder cancer is the most common urinary tumor in females, followed by kidney tumors showing slow but steady absolute and relative increase.

The presence of tumor necrosis in renal cell carcinoma (RCC) is very common and was correlated with different features including tumor size, cell type, nuclear grade as well as microvessel density and the expression of different markers. However, there are no data about the relationship between renal artery changes including atherosclerosis and fibromuscular dysplasia (FMD) and the extent of necrosis in RCC. Purpose of the study: to analyze renal arteries in patients with RCC especially regarding the thickness of medial layer and circumference of the artery using image analyzer. We analyzed a consecutive series of 57 (35 male and 22 female) patients ranging in age from 35-79 years (mean 58.9 years) who underwent nephrectomy due to RCC in the year 2003. The patients had RCC measuring from 2-16 cm (mean 7.1 cm). Renal arteries were cut during nephrectomy up to 2 cm from renal hilus. Specimens were routinely fixed, embedded in paraffin, cut and stained with hematoxylin and eosin, Mallory trichrome method and orcein. The thickness of the media of 40 patients measured from 167.86-1177.21 um (mean 571.35). Renal arteries of 17 patients (11 male, 6 female) were not measured due to technical problems. The mean thickness of media in patients with FMD was 567.33 um, those with arteriosclerosis 637.18 um and with no renal artery changes 551.11. um). There was a difference between the thicknesses of media in different types of FMD (FMD type I = 590.83 um, FMD type IIA was 476.92 um). Renal arteries of patients with arteriosclerosis were larger in comparison to arteries with no changes (4.2 mm: 3.5 mm), and control arteries obtained at autopsy, however this difference was not statistically significant. Renal arteries of patients with RCC and atherosclerotic changes have thicker media and larger diameter in comparison with controls, and those with FMD. The cause of such changes and their relationship to the degree of tumor necrosis and other changes in renal arteries should be further analyzed.

AIM: Balkan endemic nephropathy is a disease of still incompletely known etiology and is associated with increased frequency of urothelial carcinoma. Aim of the study was to compare urologic biopsies between General Hospital Dr.J.Benčević, Slavonski Brod (SB) and Sestre milosrdnice University Hospital Zagreb (ZG) in 2 five-year periods with a 20 year distance. Special point was to compare the frequency of urothelial carcinoma of the kidney and urinary bladder in endemic (SB) and non-endemic (ZG) area. MATERIALS AND METHODS: We used data bases from Department of Pathology, General Hospital Slavonski Brod and Ljudevit Jurak University Department of Pathology, Zagreb for the periods form 1980-1984 and 2000-2004. RESULTS: At the Department of Pathology SB 8770 biopsies were analyzed in the first period (1980/84), out of which 4.1% (360 cases) were urologic biopsies. At the Ljudevit Jurak University Department of Pathology 61152 biopsies were analyzed for the same period, out of which 4.8% (2919 cases) were urologic biopsies. In the second period (2000/04) total number of biopsies in SB was 27267out of which 1237 (4.5%) were from the Department of Urology, while in the same period the total number of biopsies in ZG was 73245 out of which 6599 (9%) were urologic biopsies. Out of all urologic biopsies in the period 1980/84 there were 106 (30.4%) kidney biopsies in SB and 301 (10.3%) in ZG, 65 (18%) urinary bladder biopsies in SB and 969 (33.2%) in ZG, 81 (22.5%) prostate biopsies in SB and 1072 (36%) in ZG and 5 (1.4%) testicular biopsies in SB and 216 (7.4%) in ZG. In the second period there were 248 (20%) cases of kidney biopsies in SB and 749 (11.4%) in ZG, 348 (28%) urinary bladder biopsies in SB and 1299 (19.7%) in ZG, 580 (46.7%) prostate biopsies in SB and 3382 (51.3%) in ZG and 50 cases of testicular biopsies (4%) in SB and 198 (3%) in ZG. Primary tumors of the kidney in the first period were present in 29.2 % (31 cases) out of all kidney biopsies in SB and 27% (81 cases) in ZG, and in the second period they were present in 52 % (129 cases) in SB and 54% (404 cases) in ZG. For the first period in SB urothelial carcinoma of pyelon were present in 83.8% (26 cases) out of all kidney tumors and 64.3% (83 cases) for the second period while in ZG they were present in 22.2% (18 cases) for the first period and 12.6% (51 cases) for the second period. There were 53 cases of urothelial carcinoma of the urinary bladder in SB and 630 cases in ZG for the first period, while in the second period there were 170 cases in SB and 939 in ZG. CONCLUSION: Urothelial carcinoma of the pyelon represented the majority of renal tumors in the Slavonski Brod region, probably as a consequence of BEN. The association of BEN and urothelial cancer should be analyzed further.

Arterial fibromuscular dysplasia (FMD) is a noninflammatory, nonatherosclerotic, occlusive condition of the systemic arteries, most frequently affecting renal arteries. Renal cell carcinoma (RCC) might be associated with arterial hypertension ; however, there are no data in the literature regarding the relationship between RCC and associated renal artery changes. We analyzed a consecutive series of 57 (35 male and 22 female) patients aging from 35 to 79 years (mean 58.9 years) who underwent nephrectomy due to RCC in the year 2003. The patients had RCC measuring from 2 to 16 cm (mean 7.1 cm). Specimens were routinely fixed, embedded in paraffin, cut, and stained with hematoxylin and eosin, Mallory trichrome method, and orcein. Renal arteries of 26 patients (20 male, 6 female) showed no changes. In these patients, RCC measured 2.5-11 cm in largest diameter (mean 6.6 cm). In 24 patients (10 male, 14 female), renal arteries showed FMD. RCCs in these patients measured between 2 and 16 cm (mean 8.0 cm). Seven patients had atherosclerotic changes in renal arteries. In this series, FMD was found in a significant proportion of patients with RCC, mainly in women. The cause of such changes and their relationship with RCC and systemic hypertension should be further analyzed.

INTRODUCTION AND AIM: Arterial fibromuscular dysplasia (FMD) is a condition that most frequently involves renal arteries, and is bilateral in nearly half of the cases. It is more frequent on the right side (3:1), with female predominance. FMD of the renal arteries usually involves distal parts of the artery and its branches. The aim of this study was to analyze renal arteries in patients who underwent nephrectomy due to non-tumorous conditions of the kidney. MATERIALS AND METHODS: We analyzed a consecutive series of 30 patients (m:f=14:16) who underwent nephrectomy in the year 2003, and whose histopathologic examination resulted in diagnosis of a benign condition. Eight patients were excluded from the study because specimens of renal arteries were not found. The median age of the patients (m:f=8:14) was 59.4 years (range 39-78 y). The patients were divided into the group showing FMD (group I) and those that showed no pathological changes of the renal arteries (group II). Specimens were routinely fixed, embedded in paraffin, cut at 5 μ m, stained with hematoxylin and eosin, and examined by light microscopy. RESULTS: FMD of the renal arteries (group I) was found in 10/22 patients (m:f=4:6) , with median age of 64 y (range 39-79 y). Renal arteries in 12/22 patients (group II) (m:f=4:8), with median age of 61 y (range 43-70 y), showed no changes. Histopathologic analysis of the renal tissue showed chronic pyelonephritis in all 10 patients from the group I, while 7 patients from the group II had the same diagnosis. Other diagnoses in the group II were renal calculi, cystic renal disease, hydronephrosis and renal hypoplasia. CONCLUSION: In other studies, the incidence of FMD in renal arteries, found at autopsy was about 1%, but we did not find any data about the incidence in vivo. In this study, fibromuscular dysplasia of the renal arteries was observed in a significant proportion of patients who underwent nephrectomy due to non-tumorous diseases. The significance of this finding should be further analyzed.

Aim of the study was to compare incidence between different types of urological tumors in two 5-years periods at the 20 years distance. We used histopathological data base (tHanatos) from the Department of Pathology "Ljudevit Jurak" during periods 1980/84 (first period) and 2000/04 (second period). In the first period 61152 tissue samples were analyzed, 2919 samples were obtained from urologic patients (4.8%), while in the second period there were 73245 samples analyzed of which 6599 (9.0%) were from Department of Urology. In the first period from all urological patients there were 301 (10.3%) kidney biopsies of which 81(26.9%) was due to kidney tumors while in the second period there was 749 (11.4%) kidney biopsies of which 404 (53.9%) were tumors. In the first period there were 216 (7.4%) testicular biopsies, and in the second 198 (3.0%) of which 39 (18.0%) from the first period was due to malignant neoplasms, while in the second period there were 102 (51.5%) cases of testicular tumors. From total number of Urology biopsies in 1980/84 there were 1072 (36.7%) prostatic biopsies and in 2000/04 there were 3382 (51%). Prostatic adenocarcinomas represented 15.5% (166) of all prostatic biopsies in the first period and 29.1% (983) in the second period. On the basis of this investigation we can conclude that the total number of biopsies increased, as well as the number of urological biopsies. We observed an increase in the frequency of all diagnoses of urogenital malignancies.

During the last 20 years, the occurrence of some diseases has changed due to a different way of living and new diagnostic and treatment possibilities. Diagnostic imaging (CT, MR and especially ultrasound) has increased the rate of identification of renal tumors, while there has not been any systematic screening for urologic diseases. Department of Urology is a referral center for prostate diseases since 1994. The procedure of needle core biopsy was introduced two years later and its utilization has increased significantly over the last few years. In the present study, the occurrence of some urologic diseases was compared at 20 years apart using the histopathologic database for 2 two-year periods. Data on all patients who underwent biopsies at Department of Urology during the 1980-1981 and 2000-2001 periods were included in the study. In the 1980/81 period, tissue samples of 25, 117 patients were analyzed at Department of Pathology, 1070 (4.3%) samples being obtained from urologic patients. In the 2000/01 period, there were 27, 720 patients, 2, 233 (8%) of them from Department of Urology. Urogenital tract tuberculosis was found in 13 (1.2%) patients during the 1980/81 period, and in only two (0.1%) patients in the 2000/01 period. In the 1980/81 and 2000/01 periods, there were 13 (1.2%) and 102 (4.6%) renal cell carcinomas, respectively. Of all urologic biopsies in the first period there was only one (0.1%) case, and in the 2000/01 period there were 17 (0.8%) cases of urothelial carcinoma of the pyelon. In the 1980/81 period there was not a single case of oncocytoma, whereas in the 2000/01 period 3 cases of oncocytoma were recorded. There were 106 (10%) and 64 (0.2%) testicular biopsies in the total number of urologic patients in the 1980/81 and 2000/01 periods, respectively. Of all testicular biopsies there were 13 (12%) and 29 (45%) tumors of the testis in the 1980/01 and 2000/01 periods, respectively. Seminoma accounted for 30% (n=4) and 55% (n=16) of all germ cell tumors in the 1980/81 and 2000/01 periods, respectively. Of the total number of urology biopsies performed in 1980/81 and 2000/01, there were 416 (38.8%) and 963 (43%) prostate biopsies, respectively, 59 (14 %) of them carcinomas in 1980/81 and 222 (23 %) carcinomas in 2000/01. An increasing number of prostate biopsies was recorded in the 2000/01 period, mainly due to the large proportion of needle core biopsies (304 of 966 prostatic biopsies). Prostatic biopsies accounted for 40% of all urologic biopsies in 2000 and for 46% in 2001. On the basis of this study it is concluded that the total number of urologic biopsies increased 1.9-fold comparing the first (1980-1981) and second (2000-2001) period. There was a significant increase in the number of renal cell carcinoma (3.8-fold) and urothelial carcinoma of the pyelon (8.4-fold). The total number of testicular biopsies decreased, whereas the number of testicular tumors and seminomas increased. Tuberculosis of the urogenital system decreased 13-fold.