Publications by authors named "Zoran Dzamic"

56 Publications

The Association of Polymorphisms in and Genes Involved in Redox Homeostasis in the Development and Progression of Clear Cell Renal Cell Carcinoma.

Oxid Med Cell Longev 2021 17;2021:6617969. Epub 2021 Apr 17.

Institute of Medical and Clinical Biochemistry, 11000, Serbia.

Deleterious effects of SNPs found in genes encoding transcriptional factors, as well as antioxidant and detoxification enzymes, are disputable; however, their functional significance seems to modify the risk for clear cell renal cell carcinoma (ccRCC) development and progression. We investigated the effect of specific , , gene variants and haplotype on ccRCC risk and prognosis and evaluated the association between GSTP1 and regulatory (JNK1/2) and executor (caspase-3) apoptotic molecule expression in ccRCC tissue samples and the presence of GSTP1 : JNK1/2 protein : protein interactions. Genotyping was performed in 223 ccRCC patients and 336 matched controls by PCR-CTTP and qPCR. Protein expression was analyzed using immunoblot, while the existence of GSTP1 : JNK1 protein : protein interactions was investigated by immunoprecipitation experiments. An increased risk of ccRCC development was found among carriers of variant genotypes of both rs4880 and rs1695 polymorphisms. rs6721961 genetic polymorphism in combination with both rs4880 and rs1695 showed higher ccRCC risk as well. Haplotype analysis revealed significant risk of ccRCC development in carriers of the haplotype. Furthermore, GSTP1 variant forms seem to affect the overall survival in ccRCC patients, and the proposed molecular mechanism underlying the GSTP1 prognostic role might be the presence of GSTP1 : JNK1/2 protein : protein interactions.
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http://dx.doi.org/10.1155/2021/6617969DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8068539PMC
April 2021

Polymorphisms in Genes Encoding Glutathione Transferase Pi and Glutathione Transferase Omega Influence Prostate Cancer Risk and Prognosis.

Front Mol Biosci 2021 14;8:620690. Epub 2021 Apr 14.

Faculty of Medicine, University of Belgrade, Belgrade, Serbia.

Considering the pleiotropic roles of glutathione transferase (GST) omega class members in redox homeostasis, we hypothesized that polymorphisms in and might contribute to prostate cancer (PC) development and progression. Therefore, we performed a comprehensive analysis of and SNPs' role in susceptibility to PC, as well as whether they might serve as prognostic biomarkers independently or in conjunction with other common GST polymorphisms (, , and ). Genotyping was performed in 237 PC cases and 236 age-matched controls by multiplex PCR for deletion of GST polymorphisms and quantitative PCR for SNPs. The results of this study, for the first time, demonstrated that homozygous carriers of both *A/A and *G/G variant genotypes are at increased risk of PC. This was further confirmed by haplotype analysis, which showed that H2 comprising both *A and *G variant alleles represented a high-risk combination. However, the prognostic relevance of polymorphisms in GST omega genes was not found in our cohort of PC patients. Analysis of the role of other investigated GST polymorphisms (, , and ) in terms of PC prognosis has shown shorter survival in carriers of *T/T () genotype than in those carrying at least one referent allele. In addition, the presence of *T/T genotype independently predicted a four-fold higher risk of overall mortality among PC patients. This study demonstrated a significant prognostic role of GST polymorphism in PC.
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http://dx.doi.org/10.3389/fmolb.2021.620690DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079946PMC
April 2021

Sexual dimorphism in cancer: insights from transcriptional signatures in kidney tissue and renal cell carcinoma.

Hum Mol Genet 2021 Apr;30(5):343-355

Section of Genetics, International Agency for Research on Cancer (IARC-WHO), 69372 Lyon, France.

Sexual dimorphism in cancer incidence and outcome is widespread. Understanding the underlying mechanisms is fundamental to improve cancer prevention and clinical management. Sex disparities are particularly striking in kidney cancer: across diverse populations, men consistently show unexplained 2-fold increased incidence and worse prognosis. We have characterized genome-wide expression and regulatory networks of 609 renal tumors and 256 non-tumor renal tissues. Normal kidney displayed sex-specific transcriptional signatures, including higher expression of X-linked tumor suppressor genes in women. Sex-dependent genotype-phenotype associations unraveled women-specific immune regulation. Sex differences were markedly expanded in tumors, with male-biased expression of key genes implicated in metabolism, non-malignant diseases with male predominance and carcinogenesis, including markers of tumor infiltrating leukocytes. Analysis of sex-dependent RCC progression and survival uncovered prognostic markers involved in immune response and oxygen homeostasis. In summary, human kidney tissues display remarkable sexual dimorphism at the molecular level. Sex-specific transcriptional signatures further shape renal cancer, with relevance for clinical management.
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http://dx.doi.org/10.1093/hmg/ddab031DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8098110PMC
April 2021

Role of AMPK/mTOR-independent autophagy in clear cell renal cell carcinoma.

J Investig Med 2020 12 21;68(8):1386-1393. Epub 2020 Oct 21.

Institute of Microbiology and Immunology, University of Belgrade Faculty of Medicine, Belgrade, Serbia

We examined the status and role of autophagy, a process of lysosomal recycling of cellular material, in clear cell renal cell carcinoma (ccRCC). Paired samples of tumor and adjacent non-malignant tissue were collected from 20 patients with ccRCC after radical nephrectomy. The mRNA levels of apoptosis (, , , , ) and autophagy (, , , , ) regulators were measured by RT-qPCR. The protein levels of autophagosome-associated LC3-II, autophagy receptor p62, apoptotic marker PARP, as well as phosphorylation of autophagy initiator Unc 51-like kinase 1 (ULK1), its activator AMP-activated protein kinase (AMPK) and 4EBP1, the substrate of ULK1 inhibitor mechanistic target of rapamycin (mTOR), were analyzed by immunoblotting. The mRNA levels of pro-apoptotic , anti-apoptotic and pro-autophagic , and were higher in ccRCC tumors. Autophagy induction was confirmed by an increase in phospho-ULK1 and degradation of the autophagic target p62, while apoptotic PARP cleavage was unaltered. AMPK phosphorylation was reduced and 4EBP1 phosphorylation was increased in ccRCC tissue. The expression of apoptosis regulators did not correlate with clinicopathological features of ccRCC. Conversely, high mRNA levels of and were associated with lower tumor stage, as well as with smaller tumor size and better disease-specific 5-year survival ( and ). Accordingly, low p62 protein levels, corresponding to increased autophagic flux, were associated with lower tumor stage, reduced metastasis and improved 5-year survival. These data demonstrate that transcriptional induction of autophagy in ccRCC is accompanied by AMPK/mTOR-independent increase in ULK1 activation and autophagic flux, which might slow tumor progression and metastasis independently of apoptosis.
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http://dx.doi.org/10.1136/jim-2020-001524DOI Listing
December 2020

rs1138272 Polymorphism Affects Prostate Cancer Risk.

Medicina (Kaunas) 2020 Mar 13;56(3). Epub 2020 Mar 13.

Institute of Medical and Clinical Biochemistry, 11000 Belgrade, Serbia.

: One of the most frequent genetic alterations reported to date in prostate cancer (PC) is aberrant methylation of glutathione transferase P1 (). Taking into consideration the involvement of oxidative stress in PC pathogenesis and recent advances in scientific understanding of the role of Ala114Val rs1138272 polymorphism in carcinogenesis, we hypothesized that this single-nucleotide polymorphism (SNP) influences the risk of PC independently of, or in combination with, other GST polymorphisms, including IIe105Val rs1695 or and deletion polymorphisms. : Genotyping was performed in 237 PC cases and in 236 age-matched controls by multiplex polymerase chain reaction (PCR) for deletion of GST polymorphisms and by quantitative PCR for SNPs. : We found that carriers of either *Val (rs1138272) or *Val (rs1695) variant alleles had a PC risk compared to individuals with both referent alleles (OR = 4.93, 95%CI: 2.89-8.40, < 0.001 and OR = 1.8, 95%CI: 1.19-2.73, = 0.006, respectively). Additionally, in a haplotype analysis we found that individuals with haplotype, represented by both variant alleles , had a 5.46 times higher risk of PC development compared to individuals with the most frequent haplotype (95%CI = 2.56-11.65, < 0.001), suggesting a potential role of those variants in PC susceptibility. A regression analysis on the number of risk-associated alleles per individual ( and ) showed a significant increase in the risk of developing PC, from 3.65-fold in carriers of two risk alleles (95%CI = 1.55-8.61, = 0.003) to an approximately 12-fold increase in carriers of all four risk alleles (95%CI = 3.05-44.93, < 0.001). : Prostate cancer may be influenced by multiple glutathione transferase (GST) polymorphic genes, especially , highlighting the role of gene-gene interactions in human susceptibility to this cancer.
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http://dx.doi.org/10.3390/medicina56030128DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7143748PMC
March 2020

The effect of antioxidant status on overall survival in renal cell carcinoma.

Arch Med Sci 2020 22;16(1):94-101. Epub 2019 Jul 22.

Laboratory of Molecular Biology and Endocrinology, "Vinča" Institute of Nuclear Sciences, University of Belgrade, Belgrade, Serbia.

Introduction: The oxidative stress contributes to all three phases of carcinogenesis and represents a concomitant condition in renal cell carcinoma (RCC). RCC is the most common type of neoplasm of the kidney, and despite numerous studies the set of predictive and prognostic markers of survival are still unknown. The aim of our study was to examine the relation between antioxidant (AO) status and overall survival (OS) in RCC patients.

Material And Methods: Our study included 95 patients with RCC, who underwent radical nephrectomy. We analysed the prognostic role of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glutathione, and malondialdehyde) and other clinicopathological factors (size, grade, stage, and histological subtype) on the OS of RCC patients.

Results: The 5-year OS was 54.6%. The survival analysis related to AO parameters showed no significant difference in survival of RCC patients. The concentration of malondialdehyde, an indicator of lipid peroxidation, also had no significant effect on the survival rate of RCC patients. Univariate and multivariate analysis confirmed the significance of clinicopathological parameters (size, < 0.001; Fuhrman grade, = 0.001, and stage, < 0.001) for patients' survival.

Conclusions: In our cohort of patients, different antioxidant parameters were not found to be predictors for OS of patients with RCC, who underwent radical nephrectomy.
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http://dx.doi.org/10.5114/aoms.2019.86818DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6963148PMC
July 2019

*CC Genotype (rs4925) Predicts Shorter Survival in Clear Cell Renal Cell Carcinoma Male Patients.

Cancers (Basel) 2019 Dec 17;11(12). Epub 2019 Dec 17.

Institute of Medical and Clinical Biochemistry, 11000 Belgrade, Serbia.

Omega class glutathione transferases, GSTO1-1 and GSTO2-2, exhibit different activities involved in regulation of inflammation, apoptosis and redox homeostasis. We investigated the the prognostic significance of (rs4925) and (rs156697 and rs2297235) polymorphisms in clear cell renal cell carcinoma (ccRCC) patients. GSTO1-1 and GSTO2-2 expression and phosphorylation status of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/ /mammalian target of rapamycin (mTOR) and Raf/MEK/extracellular signal-regulated kinase (ERK) signaling pathways in non-tumor and tumor ccRCC tissue, as well as possible association of GSTO1-1 with signaling molecules were also assessed. GSTO genotyping was performed by quantitative PCR in 228 ccRCC patients, while expression and immunoprecipitation were analyzed by Western blot in 30 tissue specimens. Shorter survival in male carriers of *C/C wild-type genotype compared to the carriers of at least one variant allele was demonstrated ( = 0.049). *C/C genotype independently predicted higher risk of overall mortality among male ccRCC patients ( = 0.037). Increased expression of GSTO1-1 and GSTO2-2 was demonstrated in tumor compared to corresponding non-tumor tissue ( = 0.002, = 0.007, respectively), while GSTO1 expression was correlated with interleukin-1β (IL-1β)/pro-interleukin-1β (pro-IL-1β) ratio ( = 0.260, = 0.350). Interaction of GSTO1 with downstream effectors of investigated pathways was shown in ccRCC tumor tissue. This study demonstrated significant prognostic role of polymorphism in ccRCC. Up-regulated GSTO1-1 and GSTO2-2 in tumor tissue might contribute to aberrant ccRCC redox homeostasis.
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http://dx.doi.org/10.3390/cancers11122038DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6966599PMC
December 2019

AROME-ESO Oncology Consensus Conference: access to cancer care innovations in countries with limited resources. Association of Radiotherapy and Oncology of the Mediterranean Area (AROME-Paris) and European School of Oncology (ESO - Milan).

J BUON 2019 Sep-Oct;24(5):2180-2197

Clinical Center of Montenegro, University of Montenegro, Podgorica, Montenegro.

Purpose: Cancer is a leading cause of mortality worldwide. Its incidence is still increasing, particularly in developing countries. Recent progresses further strengthen the differences between low/middle and high-income countries. This situation calls for joint action to reduce inequities in cancer outcomes among the patients. The Association of Radiotherapy and Oncology of the Mediterranean Area (AROME) and the European School of Oncology (ESO), have initiated joint conferences devoted to access to innovations in oncology in the Mediterranean area. The heterogeneity of the economic, political and cultural situations of the different participating countries, offers the opportunity to develop consensus conference.

Methods: Cancer prevention and treatment strategies were discussed according to existing international guidelines. The Scientific committee prepared 111 questions with an objective to prioritize the access to treatments and innovations in low/middle-income Mediterranean countries. The results from the votes of 65 oncology experts, coming from 16 countries and 33 institutions have been analysed and access priorities classified accordingly.

Results: Ninety six percent of the proposed general recommendations concerning national health care strategies, oncology education, and treatment organization were considered to be high priorities. Regarding access to systemic treatments, 41% of the drugs without validated predictive markers and 53% of those with validated predictive markers were considered to be 1st level priority. Only 4 biological tests were considered to be 1st level priority to access to innovation.

Conclusions: AROME-ESO consensus offers to cancer specialists from developing countries a basis for discussion with health authorities and payers on the prioritization of access to innovations in cancer care.
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April 2020

The Polyphenols as Potential Agents in Prevention and Therapy of Prostate Diseases.

Molecules 2019 Nov 4;24(21). Epub 2019 Nov 4.

Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, P.O. Box 51, 11158 Belgrade, Serbia.

In recent years, the progress of science and medicine greatly has influenced human life span and health. However, lifestyle habits, like physical activity, smoking cessation, moderate alcohol consumption, diet, and maintaining a normal body weight represent measures that greatly reduce the risk of various diseases. The type of diet is very important for disease development. Numerous epidemiological clinical data confirm that longevity is linked to predominantly plant-based diets and it is related to a long life; whereas the western diet, rich in red meat and fats, increases the risk of oxidative stress and thus the risk of developing various diseases and pre-aging. This review is focused on the bioavailability of polyphenols and the use of polyphenols for the prevention of prostate diseases. Special focus in this paper is placed on the isoflavonoids and flavan-3-ols, subgroups of polyphenols, and their protective effects against the development of prostate diseases.
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http://dx.doi.org/10.3390/molecules24213982DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6864651PMC
November 2019

The assessment of knowledge and attitudes of men in Serbia about prostate cancer and possibilities for its early detection and treatment in 2011: a cross-sectional study.

J BUON 2019 Jan-Feb;24(1):233-238

Clinic for Urology, Clinical Center of Serbia, Belgrade, Serbia.

Purpose: To assess the knowledge and attitudes of men in Serbia about prostate cancer (PCa) and possibilities for its early detection and treatment in 2011.

Methods: This cross-sectional study included 407 men of various ages and education levels selected randomly and divided in 2 groups according to age (up to 40 and over 40 years). The assessment of knowledge and attitudes was based on a survey made up of 12 multiple choice questions conducted with direct contact with respondents from October 15th to December 15th 2011 with their voluntary consent. The results were evaluated in the total sample and between the groups.

Results: Patient groups significantly differed according to knowledge about PCa treatment success (p<0.001) and stage in which PCa is most frequently detected (p<0.001) as well as according to attitudes about community-based interventions for increasing the awareness of PCa (p<0.001). Sixty-one percent of respondents over 50 years hadn't done preventive prostate examination despite recommendations. Ninety percent of all respondents believed the community-based intervention should have been implemented in Serbia to increase the men's awareness of PCa.

Conclusion: The study reported lack of men's knowledge about PCa in Serbia in 2011, while there was a common agreement among men on the necessity of spreading more information about this disease.
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August 2019

Urothelial carcinoma of the upper urinary tract: preoperative pyuria is not correlated with bladder cancer recurrence and survival.

Int Urol Nephrol 2019 May 1;51(5):831-838. Epub 2019 Apr 1.

Institute of Epidemiology, Faculty of Medicine, University of Belgrade, Belgrade, Serbia.

Objective: To identify the impact of preoperative pyuria on the bladder cancer recurrence and survival of patients who were treated surgically for UTUC.

Patients And Methods: Study included 319 consecutive patients who were treated with RNU for UTUC. Cox proportional hazard regression models were used to evaluate the association of preoperative pyuria with outcome.

Results: Eighty patients (25.1%) had pyuria. Preoperative pyuria was associated with sex (P = 0.01), tumor focality (P = 0.01), tumor size (P = 0.05), tumor stage (P = 0.01), lymph node metastasis (P = 0.01), lymphovascular invasion (P = 0.02), and chemotherapy (P = 0.04). A total of 102 patients recurred, with a median time to bladder recurrence of 24.2 months. Bladder cancer recurrence-free survival rates for these 319 patients at 1, 3, 5, 7, and 10 years were 84.6, 72.4, 69.0, 68.3, and 68.0%, respectively. Preoperative pyuria was not independently associated with bladder cancer recurrence (HR 1.15; p = 0.5). Preoperative pyuria was associated with OS (HR 1.57; p = 0.02) and CSS (HR 1.65; p = 0.02). However, preoperative pyuria was not independently associated with OS and CSS (HR 1.07; p = 0.79).

Conclusions: Preoperative pyuria is unable to predict outcomes in a single-centre series of consecutive patients who were treated with RNU.
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http://dx.doi.org/10.1007/s11255-019-02133-1DOI Listing
May 2019

HTERT promoter methylation and single nucleotide polymorphism (-245 T>C) affect renal cell carcinoma behavior in Serbian population.

J BUON 2018 Nov-Dec;23(6):1887-1892

Institute of Pathology, School of Medicine, University of Belgrade, Belgrade, Serbia.

Purpose: Renal cell carcinoma (RCC) is the most common renal cancer in adults and includes several subtypes that may be distinguished by their histology, genetic background, clinical course and responses to treatment. Human telomerase reverse transcriptase (hTERT), a crucial enzyme for telomere maintenance, has been linked to RCC development. The purpose of this study was to search for genetic and epigenetic alterations in hTERT (promoter mutations and methylation and gene amplification), and to establish a possible association between molecular and clinico-pathological characteristics of RCC.

Methods: DNA was extracted from 31 formalin-fixed, paraffin-embedded tumor samples and 23 blood samples from 54 patients with RCC. Polymerase chain reaction (PCR) products were sequenced and analyzed using the Sequencher software. HTERT amplification was determined by quantitative PCR, while the promoter methylation status was assessed by methylation specific PCR. Statistical analysis was performed using SPSS.

Results: No mutations could be detected in the hTERT promoter but only a single nucleotide polymorphism (SNP) (-245 T>C). In 54 analyzed RCC cases, the variant allele C was present in homozygous or heterozygous form in 48% of the patients. The C allele was significantly more frequent in low grade tumors (p=0.046). Gene amplification was detected in 19.4% of the 31 RCCs and hTERT methylation in 54.8% of the 31 samples. An association was established between methylation and histological type of RCC (p=0.047).

Conclusions: HTERT seems to be implicated in RCC pathogenesis since the promoter polymorphism exerts a modulation effect on tumor behavior. In addition, hTERT promoter methylation status is related to RCC histology.
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August 2019

Association between GPX1 and SOD2 genetic polymorphisms and overall survival in patients with metastatic urothelial bladder cancer: a single-center study in Serbia.

J BUON 2018 Jul-Aug;23(4):1130-1135

Clinic of Urology, Clinical Center of Serbia, Belgrade, Serbia

Purpose: Purpose: Urothelial bladder cancer (UBC) is the most common malignancy of urinary tract in the developed world. In metastatic UBC, systemic chemotherapy still remains the mainstay of initial treatment. Inter-individual differences in treatment outcome partially may be the consequence of genetic variations in enzymes that modulate oxidative stress. Therefore, we aimed to determine the potential prognostic role of single nucleotide polymorphism (SNP) of the two antioxidant enzymes glutathione peroxidase 1 (GPX1) and superoxide dismutase 2 (SOD2) in metastatic UBC patients treated with cisplatin-based chemotherapy.

Methods: Methods: This prospective single-center hospital-based case-control study included 33 patients with metastatic UBC treated with cisplatin-based chemotherapy and 227 healthy controls. GPX1 SNP (rs1050450) was assessed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and SOD2 SNP (rs4880) was determined by quantitative PCR (q-PCR). Overall survival (OS) was evaluated using Kaplan–Meier survival analysis during 2-year follow up period, with the log-rank test for prognostic significance.

Results: Results: No significant difference was observed in the distributions of GPX1 and SOD2 gene variants between patients and controls (p˃0.05). Regarding GPX1 polymorphism, no impact of GPX1 polymorphism on OS could be demonstrated (p˃0.05). Finally, Kaplan-Meier survival analysis showed no association between SOD2 polymorphism and OS (p˃0.05).

Conclusions: Conclusions: No association was found between polymorphism of GPX1 and SOD2 and OS in patients with metastatic urothelial bladder cancer treated with cisplatin-based chemotherapy.
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September 2019

Immunohistochemical expression of proliferative markers in renal cell carcinoma.

J BUON 2018 Jul-Aug;23(4):1103-1110

Institute of Pathology, University of Belgrade School of Medicine, Belgrade, Serbia.

Purpose: The purpose of this study was to investigate into the expression of cyclin A and telomerase in renal cell carcinoma (RCC) and to analyze the relationship between expression and the clinicopathological characteristics of the tumor and their impact on survival.

Methods: The overall material included 74 samples of RCC and 4 of normal renal tissue. Primary cyclin A antibody from Santa Cruz Biotechnology and TERT MA5-16034 antibody from Thermo Fisher Scientific Inc were used. Staining was performed by streptavidin-biotin technique using DAKO LSAB+ kit. Statistical analyses were performed using of SPSS 23 Statistics software from IBM.

Results: No differences in cyclin A and telomerase expression among gender and age groups were found, nor did the tumor dimensions have any significant impact on expression. Also, tumor grades and stages did not differ. However, histological types differed in favor of the papillary type. A significant positive correlation between both markers, as well as between the expression and tumor stage and grade was noticed. Only the tumor stage had negative impact on survival.

Conclusions: Although not affecting survival, the expression of cyclin A and telomerase increased with tumor stage and grade, suggesting that cyclin A and telomerase could be potential proliferative immunohistochemical markers of RCC.
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September 2019

Concomitance of Polymorphisms in Glutathione Transferase Omega Genes Is Associated with Risk of Clear Cell Renal Cell Carcinoma.

Tohoku J Exp Med 2018 09;246(1):35-44

Institute of Medical and Clinical Biochemistry, Faculty of Medicine, University of Belgrade.

Glutathione S-transferases (GSTs), a superfamily of multifunctional enzymes, play an important role in the onset and progression of renal cell carcinoma (RCC). However, novel GST omega class (GSTO), consisting of GSTO1-1 and GSTO2-2 isoenzymes, has not been studied in RCC yet. Two coding single nucleotide polymorphisms (SNPs) supposedly affect their functions: GSTO1*C419A (rs4925) causing alanine to aspartate substitution (*A140D) and GSTO2*A424G (rs156697) causing asparagine to aspartate substitution (*N142D), and have been associated with several neurodegenerative diseases and cancers. Functional relevance of yet another GSTO2 polymorphism, identified at the 5' untranslated (5'UTR) gene region (GSTO2*A183G, rs2297235), has not been clearly discerned so far. Therefore, we aimed to assess the effect of specific GSTO1 and GSTO2 gene variants, independently and in interaction with established risk factors (smoking, obesity and hypertension) on the risk for the most aggressive RCC subtype, the clear cell RCC (ccRCC). Genotyping was performed in 239 ccRCC patients and 350 matched controls, while plasma levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage, were determined by ELISA. As a result, combined effect of all three variant genotypes exhibited almost 3-fold risk of RCC development. Additionally, this association was confirmed at the haplotype level [variant GSTO1*A/GSTO2*G (rs156697)/GSTO2*G (rs2297235) haplotype], suggesting a potential role of those variants in propensity to RCC. Regarding the gene-environment interactions, variant GSTO2*G (rs156697) homozygous smokers are at higher ccRCC risk. Association in terms of oxidative DNA damage was found for GSTO2 polymorphism in 5'UTR and 8-OHdG. In conclusion, the concomitance of GSTO polymorphisms may influence ccRCC risk.
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http://dx.doi.org/10.1620/tjem.246.35DOI Listing
September 2018

Clinicopathological characteristics and survival in Serbian patients with renal cell carcinoma: a retrospective analysis.

J BUON 2017 Nov-Dec;22(6):1434-1440

Laboratory of Molecular Biology and Endocrinology, "Vinca" Institute of Nuclear Sciences, University of Belgrade, Belgrade, Serbia.

Purpose: Indications of kidney cancer outcome in lowerincome countries are based on an incidence/mortality ratio due to lack of survival information. This study was conducted to provide outcome data in Serbian patients with renal cell carcinoma (RCC) and to identify prognostic factors that could affect their overall survival (OS).

Methods: This retrospective study included 185 patients who underwent nephrectomy. We assessed certain clinicopathological data including age, gender, tumor size, grade, stage and histological subtypes for their possible impact on OS.

Results: The 5-year OS was 63.2%. Significant association was found between OS and age (log-rank 12.455, p=0.006), tumor size (log-rank 26.425, p=0.000), grade (log-rank 13.249, p=0.000) and stage (log-rank 43.235, p=0.000). Univariate analysis indicated size (p=0.000), grade (p=0.001) and stage (p=0.000) as prognostic factors for OS. In multivariate analysis, grade (p=0.014) and stage (p=0.000) remained significant predictors of OS.

Conclusion: Tumor grade and stage were identified as independent prognostic factors of OS survival in Serbian patients with RCC.
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July 2019

Immunohistochemical study of cyclin A and p16 expression in patients with renal cell carcinoma.

J BUON 2017 Sep-Oct;22(5):1322-1327

The Institute of Pathology, School of Medicine, University of Belgrade, Belgrade, Serbia.

Purpose: Renal cell carcinoma (RCC) is the most common malignant kidney tumor in adults. Dysregulation of the cell cycle can lead to cancer development. In this study, the mitosis-associated cyclin A and p16, a negative controller, were investigated as potential key points in the RCC development.

Methods: This retrospective study included 74 patients with RCC. The expression of cyclin A and p16 and their correlation to histopathological parameters (TNM stage, histological subtype, nuclear grade, tumor size), gender, age, and clinical outcome were studied and analyzed.

Results: The highest median value for cyclin A (40%; range 0-70)) and for p16 (57.5%); range 35-80) were found in the papillary histological subtype. Survival analysis showed that in the group of patients that had died before September 2015, the median value for cyclin A was 20% (range 0-60), which was significantly higher than 5% (range 0-70), found in the group of patients that survived (p=0.019).

Conclusions: In relation to the histological subtype, the papillary type of RCC was associated with a significantly higher expression of cyclin A and p16 compared to other subtypes of RCC. High expression of cyclin A indicated worse prognosis, therefore cyclin A could be considered to be a significant prognostic marker.
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July 2019

Genome-wide association study identifies multiple risk loci for renal cell carcinoma.

Nat Commun 2017 06 9;8:15724. Epub 2017 Jun 9.

Clinical Center of Serbia (KCS), Clinic of Urology, University of Belgrade-Faculty of Medicine, 11000 Belgrade, Serbia.

Previous genome-wide association studies (GWAS) have identified six risk loci for renal cell carcinoma (RCC). We conducted a meta-analysis of two new scans of 5,198 cases and 7,331 controls together with four existing scans, totalling 10,784 cases and 20,406 controls of European ancestry. Twenty-four loci were tested in an additional 3,182 cases and 6,301 controls. We confirm the six known RCC risk loci and identify seven new loci at 1p32.3 (rs4381241, P=3.1 × 10), 3p22.1 (rs67311347, P=2.5 × 10), 3q26.2 (rs10936602, P=8.8 × 10), 8p21.3 (rs2241261, P=5.8 × 10), 10q24.33-q25.1 (rs11813268, P=3.9 × 10), 11q22.3 (rs74911261, P=2.1 × 10) and 14q24.2 (rs4903064, P=2.2 × 10). Expression quantitative trait analyses suggest plausible candidate genes at these regions that may contribute to RCC susceptibility.
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http://dx.doi.org/10.1038/ncomms15724DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5472706PMC
June 2017

Testosterone and dihydrotestosterone levels in the transition zone correlate with prostate volume.

Prostate 2017 Jul 8;77(10):1082-1092. Epub 2017 Jun 8.

Clinic of Urology, Clinical Center of Serbia, Belgrade, Serbia.

Background: There is still no consensus regarding intraprostatic androgen levels and the accumulation of androgens in the hyperplastic prostatic tissue. The current opinion is that intraprostatic dihydrotestosterone (DHT) concentrations are maintained but not elevated in benign prostatic hyperplasia (BPH), while there is no similar data concerning intraprostatic testosterone (T).

Methods: Tissue T (tT) and tissue DHT (tDHT) concentration were determined in 93 patients scheduled for initial prostate biopsy. The criteria for biopsy were abnormal DRE and/or PSA > 4 ng/mL. Total prostate volume (TPV) was determined by transrectal ultrasound (TRUS). During TRUS- guided prostate biopsy, 10-12 samples were collected from the peripheral zone (PZ) and two additional samples were collected from the transition zone (TZ). The samples from the TZ were immediately frozen in liquid nitrogen at -70°C, and transported for tissue androgen determination, using liquid chromatography mass spectrometry (LC-MS).

Results: Pathological analysis revealed that prostate cancer (PCa) was present in 45 and absent in 48 patients. In the whole group, there were 42 men with small prostate (TPV < 30 mL) and 51 with enlarged prostate (TPV ≥ 31 mL). The overall average tT level was 0.79 ± 0.66 ng/g, while the average tDHT level was 10.27 ± 7.15 ng/g. There were no differences in tT and tDHT level in prostates with and without PCa. However, tT and tDHT levels were significantly higher in larger, than in smaller prostates (tT: 1.05 ± 0.75 and 0.46 ± 0.29 ng/g, and tDHT: 15.0 ± 6.09 and 4.51 ± 2.75 ng/g, respectively). There were strong correlations between tT and TPV (r = 0.71), and tDHT and TPV (r = 0.74).

Conclusions: The present study confirmed that both T and DHT accumulated in the stroma of enlarged prostates; the degree of accumulation correlated with prostate volume.
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http://dx.doi.org/10.1002/pros.23365DOI Listing
July 2017

GSTM1 genotype is an independent prognostic factor in clear cell renal cell carcinoma.

Urol Oncol 2017 06 9;35(6):409-417. Epub 2017 Mar 9.

Institute of Medical and Clinical Biochemistry, Belgrade, Serbia; Faculty of Medicine, University in Belgrade, Belgrade, Serbia. Electronic address:

Purpose: Owing to dual functionality of cytosolic glutathione S-transferases (GSTs), they might affect both the development and the progression of renal cell carcinoma (RCC). However, the data on the prognostic value of GST polymorphism in patients with RCC are scarce. Hence, we evaluated the effect of GST gene variants on both the risk of RCC development and the postoperative prognosis in patients with clear cell RCC (ccRCC).

Methods: GST genotypes were determined in 305 patients with RCC and 326 matched controls, whereas the overall survival was evaluated in patients with ccRCC only. The presence of GSTM1:ASK1 protein-protein interaction in ccRCC tissue samples was analyzed by methods of immunoprecipitation and immunoblot.

Results: We noted an increased risk of RCC development in carriers of GSTM1-null and GSTP1-variant genotype (P<0.05). On the contrary, survival analysis indicated shorter overall survival for patients with ccRCC with GSTM1-active genotype (P = 0.026). Furthermore, patients with ccRCC with GSTM1-active genotype had significantly higher hazard ratio (P<0.05), in analyzed regression models, compared with the carriers of GSTM1-null genotype. Finally, the presence of GSTM1:ASK1 protein-protein interaction was found in all RCC tissue samples studied.

Conclusions: Carriers of GSTM1-null and GSTP1-variant genotypes are in increased risk of RCC development. On the contrary, GSTM1-null genotype is associated with favorable postoperative prognosis in ccRCC. The possible molecular mechanism underlying the role of GSTM1 protein in RCC progression might be the presence of GSTM1:ASK1 protein-protein interaction. Hence, determination of GSTM1-genotype might serve as a valuable indicator in both RCC risk assessment and postoperative prognosis.
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http://dx.doi.org/10.1016/j.urolonc.2017.02.005DOI Listing
June 2017

Partial versus radical nephrectomy for pT1a renal cancer in Serbia.

J BUON 2016 Nov-Dec;21(6):1449-1453

Clinic of Urology, Clinical Center of Serbia, Belgrade, Serbia.

Purpose: To assess the treatment outcome of pT1a renal tumors, comparing overall survival (OS) in patients treated with radical nephrectomy (RN) and partial nephrectomy (PN), and to examine the rate of utilization of PN in a tertiary institution in Serbia.

Methods: Included were patients treated for pT1a kidney tumors with open RN or open PN during 1996-2013. The inclusion criterion was the pathological tumor stage T1a. Exclusion criteria were higher pathological stages, metastatic presentation, or imperative indications for partial nephrectomy. Patients were followed-up every 3 to 4 months for the first year after surgery, every 6 months until the 5th year, and annually thereafter.

Results: 286 patients were included in the study, and PN was performed in 177 (61.9%) of them, whereas RN was performed in the remaining 109 (38.1%). The median follow- up for the entire group was 42.0 months (interquartile range 74.5). There were no statistically significant differences between groups in cancer-specific survival (CSS) (log-rank=0.506; p=0.477). Patients selected for RN were more likely to be older, symptomatic at presentation, and have larger tumors. There was no statistically significant difference in OS between the two groups (log-rank=2.616; p=0.106). In 1996, 20% of the patients were treated with PN; this number increased to 88% in 2013.

Conclusion: We did not find OS advantage for PN compared to RN in the setting of a developing country. The use of PN is increasing and is now utilized for ∼90% of pT1a renal tumors.
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June 2017

Combined GSTM1-Null, GSTT1-Active, GSTA1 Low-Activity and GSTP1-Variant Genotype Is Associated with Increased Risk of Clear Cell Renal Cell Carcinoma.

PLoS One 2016 8;11(8):e0160570. Epub 2016 Aug 8.

Institute of Medical and Clinical Biochemistry, Belgrade, Serbia.

The aim of this study was to evaluate specific glutathione S-transferase (GST) gene variants as determinants of risk in patients with clear cell renal cell carcinoma (cRCC), independently or simultaneously with established RCC risk factors, as well as to discern whether phenotype changes reflect genotype-associated risk. GSTA1, GSTM1, GSTP1 and GSTT1 genotypes were determined in 199 cRCC patients and 274 matched controls. Benzo(a)pyrene diolepoxide (BPDE)-DNA adducts were determined in DNA samples obtained from cRCC patients by ELISA method. Significant association between GST genotype and risk of cRCC development was found for the GSTM1-null and GSTP1-variant genotype (p = 0.02 and p<0.001, respectively). Furthermore, 22% of all recruited cRCC patients were carriers of combined GSTM1-null, GSTT1-active, GSTA1-low activity and GSTP1-variant genotype, exhibiting 9.32-fold elevated cRCC risk compared to the reference genotype combination (p = 0.04). Significant association between GST genotype and cRCC risk in smokers was found only for the GSTP1 genotype, while GSTM1-null/GSTP1-variant/GSTA1 low-activity genotype combination was present in 94% of smokers with cRCC, increasing the risk of cRCC up to 7.57 (p = 0.02). Furthermore, cRCC smokers with GSTM1-null genotype had significantly higher concentration of BPDE-DNA adducts in comparison with GSTM1-active cRCC smokers (p = 0.05). GSTM1, GSTT1, GSTA1 and GSTP1 polymorphisms might be associated with the risk of cRCC, with special emphasis on GSTM1-null and GSTP1-variant genotypes. Combined GSTM1-null, GSTT1-active, GSTA1 low activity and GSTP1-variant genotypes might be considered as "risk-carrying genotype combination" in cRCC.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0160570PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4976979PMC
August 2017

Pulsed versus continuous mode fluoroscopy during PCNL: safety and effectiveness comparison in a case series study.

Urolithiasis 2016 Nov 28;44(6):565-570. Epub 2016 Apr 28.

2nd Urology Department, Athens Medical School, Sismanogleio General Hospital, University of Athens, Athens, Greece.

To compare the total fluoroscopy time (FT) based on the fluoroscopy mode used-continuous vs. pulsed-in patients who underwent percutaneous nephrolithotomy (PCNL). The study cohort evaluated 111 patients who underwent PCNL by a single surgeon. Standard (continuous) fluoroscopy of 30 frames per second (fps) was used in the first 56 cases (SF group), while the next 55 consecutive cases were performed under pulsed fluoroscopy of two fps (PF group). The presence of surgeon's previous experience decreased the possible impact of the learning curve on the outcome. In both groups, using ultrasound in combination to fluoroscopy performed the renal access. The stone complexity was determined using Guy's stone score (GSS). Complications were evaluated using Clavien-Dindo classification. Median FT was significantly lower in PF group (76.8 s) compared to SF group (155.4 s) (p < 0.001). Stone-free rate was related to the Guy's stone score (GSS) classification reaching 100 % in GSS 1 cases in both groups. In GSS 2 cases the stone free rate was 87.5 % in SF group, while in PF group it was 92.3 %. Stone free rate in GSS 3 cases was 73.3 and 85.7 % in SF and PF groups, respectively. In cases of GSS 4 stone free rate was 52 % in SF group and 55.6 % in PF group, respectively. Presence of residual fragments and complications were comparable in both groups. Following ultrasound-guided puncture during PCNL, the use of pulsed fluoroscopy leads to significantly lower radiation exposure comparing to the use of continuous fluoroscopy. This advantage does not compromise the safety and efficacy of the procedure.
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http://dx.doi.org/10.1007/s00240-016-0885-6DOI Listing
November 2016

Acute Renal Failure in Different Malignant Tumors.

Curr Med Chem 2016 ;23(19):2041-6

Department of Pathology, Faculty of Medicine, University of Belgrade, Serbia.

Acute renal failure (ARF) represents a severe complication of malignancies, that causes significant morbidity and mortality. ARF is a common part of multiple organ dysfunction in critically ill patients with cancer with reported mortality rates from 72% to 85% in patients who need renal replacement therapy. The pathways leading to ARF in cancer patients are common to the development of ARF in other conditions. However, certain factors leading to the development of ARF may be associated to the tumor or to the tumor therapy. The purpose of this review is to give specific aspects of renal disease in critically ill cancer patients (CICPs), to overview the causes of ARF in CICPs and to describe recent progress in the management of these complications, including treatment toxicity and bone marrow transplantation (BMT). The prevention of ARF is obligatory and therefore the possible treatments of ARF in CICPs are also discussed.
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http://dx.doi.org/10.2174/0929867323666160407113245DOI Listing
February 2017

Primary dorsal buccal mucosa graft urethroplasty for anterior urethral strictures in patients with lichen sclerosus.

Int Urol Nephrol 2016 Apr 11;48(4):541-5. Epub 2016 Jan 11.

Sava Perovic Foundation (RD), Belgrade, Serbia.

Purpose: To report our ongoing experience with dorsal buccal mucosa graft (BMG) urethroplasty for the primary repair of anterior urethral strictures in patients with lichen sclerosus (LS).

Patients And Methods: A total of 32 men with LS underwent BMG urethroplasty from January 2010 to September 2012. In 27 patients, stricture was limited to the penile urethra, while in five patients, both bulbar and penile urethra were involved. In these five patients, the entire anterior urethra was replaced with BMG. In nine (28.1%) younger patients (mean age 38.2 years, range 33-45), with adverse local conditions and significant scarring, two-stage repair was done. The paired t test was performed on preoperative and postoperative Qmax as well as on preoperative and postoperative post-void residual urine volume, and the Fisher exact test was used to assess success between treatment groups. The chi-squared test was used to compare categorical data.

Results: The overall success rate was 90.6%. Complications occurred in 9.4% of the patients (3 of 32) including hematoma in two patients and fistula in one patient. In this cohort of patients, mean preoperative Qmax was 6.2 ml per second (range 2.6-10.2) versus 18.2 (range 15.8-21.2) postoperatively (at 9 months), which was statistically significant (p < 0.002). Also, mean preoperative post-void residual urine volume was 110 ml (range 75-180) versus 19 ml (range 10-40) postoperatively at 9 months, which was statistically significant (p < 0.004).

Conclusion: Buccal mucosa is the most reliable graft for repairing anterior urethral strictures in patients with LS. Minimal complications are observed, even in cases of long stenosis completely afflicting anterior urethra.
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http://dx.doi.org/10.1007/s11255-015-1202-yDOI Listing
April 2016

Idionathic retroneritoneal fibrosis: A Report on 15 Patients.

Vojnosanit Pregl 2015 Oct;72(10):928-31

Background/aim: Retroperitoneal fibrosis (RPF) repre sents a chronic pathological process characterized by fibro sis which entraps and compresses the ureters and the grea blood vessels in the retroperitoneal space. A specific form of RPF is idiopathic RPF, an uncommon collagen vasculaj disease of unclear etiology. The series of 15 patients whicl underwent open surgical repair due to idiopathic RPF is presented herein.

Methods: From 1989 to 2012, 11 male and 4 female patients underwent surgery due to primar RPF. The ureters were entrapped unilaterally (7 patients), or bilaterally (8 patients). Major symptoms included low bacl pain due to hydronephrosis (9 patients), uremia (4 patients) and urinary tract infection (2 patients). The diagnosis was based on intravenous urography (IVU), retrograde uretero pyelography and computed tomography (CT).

Results: Surgical procedures included intraperitoneal ureteral dis placement (8 patients) and ureteral wrapping with omental flap (6 patients). One patient underwent bilateral uretera stenotic segments resection and oblique ureterography, followed by wrapping with omental flap. Pathological examination confirmed primary RPF in all patients. The meat operative time was 3.5 h (range 2.5-4.5 h). The average in trahospital stay was 21 days (range 16-26 days). The meat follow up was 32 months (6-46 months). During the follow up, 12 patients had improvement on IVU.

Conclusion: Early recognition of signs and symptoms of RPF is of the utmost importance for the outcome. Surgical procedures including ureteral wrapping with omental flap, or intraperi toneal ureteral displacement, usually represent definitive treatment.
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http://dx.doi.org/10.2298/vsp140426076hDOI Listing
October 2015

Urothelial carcinoma: Recurrence and risk factors.

J BUON 2015 Mar-Apr;20(2):391-8

Clinic of Urology, Clinical Center of Serbia, School of Medicine, University of Belgrade, Belgrade, Serbia.

Urothelial carcinomas are malignant tumors that arise from the urothelial epithelium and may involve the lower and upper urinary tract. They are characterized by multiple, multifocal recurrences throughout the genitourinary tract. Bladder tumors account for 90-95% of urothelial carcinomas and are the most common malignancies of the urinary tract. Upper urinary tract urothelial carcinomas (UTUC) are relatively rare, accounting for 5% of urothelial tumors. The incidence of subsequent bladder cancer after surgical treatment for UTUC is approximately 15-50%. In contrast, patients with a primary tumor of the bladder have a low risk (2-6%) the development of UTUC. Identification of prognostic factors and early detection of recurrent disease provide a better strategy for postoperative monitoring, surveillance, and potentially improve patient outcomes. In this review study we discuss the main risk factors for UTUC recurrence after radical cystectomy, and risk factors for intravesical recurrence after radical nephroureterectomy.
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July 2015

Prognostic Impact of Preoperative Anemia on Urothelial and Extraurothelial Recurrence in Patients With Upper Tract Urothelial Carcinoma.

Clin Genitourin Cancer 2015 Oct 30;13(5):485-91. Epub 2015 Mar 30.

Institute of Epidemiology, School of Medicine, University of Belgrade, Belgrade, Serbia.

Background: To investigate the prognostic impact of preoperative anemia on urothelial and extraurothelial recurrence after radical nephroureterectomy.

Methods: A single-center series of 238 consecutive patients who were treated with radical nephroureterectomy for upper tract urothelial carcinoma was evaluated. We categorized patients on the basis of hemoglobin level into 2 groups, including normal or anemia. Survival was estimated using the Kaplan-Meier method. Cox proportional hazard regression models were used to evaluate the association of preoperative anemia with outcome, controlling for clinicopathologic variables.

Results: Ninety-seven patients (40.8%) had anemia (median hemoglobin level, 143 vs. 107 g/L). Preoperative anemia was associated with history of bladder cancer (P = .01), tumor multifocality (P = .03), lymphovascular invasion (P = .05), and adjuvant chemotherapy (P = .01). Higher tumor stage and grade, and lymph node metastasis were significantly associated with preoperative anemia. Preoperative anemia was independently associated with extraurothelial recurrence (hazard ratio, 1.95; 95% confidence interval, 1.14-3.34; P = .01) in multivariate Cox regression analyses. Only a history of bladder tumor (hazard ratio, 2.07; P = .009) and tumor multifocality (hazard ratio, 3.97; 95% confidence interval, 2.37-6.67; P < .001) were independently associated with urothelial recurrence. The 5-year cancer-specific survival for patients with normal hemoglobin level was 82.1% and for patients with preoperative anemia was 54.2%.

Conclusion: Patients with preoperative anemia had a greater probability of having upper tract urothelial carcinoma with higher tumor stages, higher tumor grades, and lymph node metastasis (pN+). Preoperative anemia was statistically significantly associated with worse cancer-specific survival and extraurothelial recurrence in patients who underwent radical nephroureterectomy.
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http://dx.doi.org/10.1016/j.clgc.2015.03.007DOI Listing
October 2015

Bilateral triple renal pelvis: a case report.

Vojnosanit Pregl 2015 Feb;72(2):185-7

Introduction: Triple renal pelvis is an extremely rare variation of the renal collecting system. To the authors' knowledge, bilateral triple renal pelvis has not yet been described in the literature.

Case Report: A 55-year-old man was hospitalized due to papillary bladder cancer, detected on ultrasonography. As incidental finding, intravenous urography revealed bilateral triple renal pelvis. Six weeks after transurethral resection of the bladder tumor (TURB) the patient was admitted again, for the second TURB. Computed to mography-urography confirmed the presence of bilateral triple renal pelvis.

Conclusion: The unique case of bilateral triple renal pelvis was presented as an extremely rare variation of the renal collecting system.
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http://dx.doi.org/10.2298/vsp1502185pDOI Listing
February 2015