Immune analysis of urine and plasma samples from patients with clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the third most common type of urological malignancy worldwide, and it is associated with a silent progression and late manifestation. Patients with a metastatic form of ccRCC have a poor prognosis; however, when the disease is diagnosed early, it is largely curable. Currently, there are no biomarkers available in clinical practice for ccRCC. Thus, the aim of the present study was to measure 27 biologically relevant cytokines in preoperative and postoperative urine samples, and in preoperative plasma samples from 34 patients with ccRCC, and to evaluate their diagnostic significance. The concentrations of cytokines were assessed by multiplex immune assay. The results showed significantly higher levels of IL-1 receptor antagonist, IL-6, IL-15, chemokine (C-C motif) ligand (CCL)2, CCL3, CCL4, C-X-C motif ligand (CXCL)10, granulocyte-macrophage colony stimulating factor (GM-CSF) and platelet-derived growth factor-BB (PDGF-BB), and lower levels of granulocyte colony stimulating factor (G-CSF) in urine samples from patients prior to surgery compared with those in the controls. Notably, the urine levels of G-CSF, IL-5 and vascular endothelial growth factor differed following tumor removal compared with the preoperative urine levels. In addition, urinary G-CSF, GM-CSF, IL-6, CXCL10, CCL5 and PDGF-BB appeared to be potential markers of tumor grade. Plasma from patients with ccRCC contained significantly higher levels of IL-6 and lower levels of CCL2 than control plasma. In conclusion, the present findings indicated that urinary and circulating cytokines may represent a promising novel tool for the early diagnosis of ccRCC and/or prediction of tumor grade.

The effect of pelvic floor muscle training in men with benign prostatic hyperplasia and overactive bladder.

BACKGROUND: Men with overactive bladder (OAB) and benign prostatic hyperplasia (BPH), will have deterioration in the quality of life. OBJECTIVE: The aim of this study was to evaluate the effect of combining pelvic floor muscle training with the urgency suppression technique (PFMT-st) and silodosin in comparison with silodosin in men with benign prostatic hyperplasia (BPH) and overactive bladder (OAB) after 12 weeks of treatment. PATIENTS AND METHODS: A total of 158 patients were randomized into two groups. The control group received oral silodosin at a daily dose of 8 mg. The experimental group was administered PFMT-st and silodosin. The evaluation methods included the number of voids and intensity of urgencies over 24 h using a micturition diary, the International Prostate Symptom Score (IPSS), the Overactive Bladder Questionnaire (OAB-q), and the patient global impression of improvement (PGI-I). RESULTS: 142 of 172 (86.6%) men were assessed (70 in the control group, 72 in the experimental group). The significant changes were in favor of the experimental group (p < 0.001) in the number of voids per 24 h (- 1.95 ± 1.94 vs. - 0.90 ± 1.44), the OAB-q symptom score (- 14.25 ± 10.05 vs. - 9.28 ± 10.60), the intensity of urgencies (- 0.97 ± 0.53 vs. 0.24 ± 0.57), the IPSS (- 4.59 ± 3.00 vs. - 2.30 ± 3.63), and in the PGI-I (2.24 ± 0.79 vs. 3.60 ± 0.92). CONCLUSIONS: The addition of PFMT-st to silodosin treatment significantly improved OAB in men with BPH. This is the first study to confirm that PFMT-st should be the first-choice treatment for OAB in BPH.

Immunobiochemical profile of clear cell renal cell carcinoma (ccRCC): A preliminary study.

Clear cell renal cell carcinoma (ccRCC) is the most common variant of RCC. It is an aggressive disease with an unfavorable prognosis. The rich immune infiltrates present in the tumor microenvironment (TME) of ccRCC produce various signaling molecules, especially cytokines, which primarily activate the Jak/STAT pathway and significantly influence tumor pathogenesis. STAT3 has a well-defined oncogenic character. Using multiplex assays and ELISA, we have measured the concentrations of 27 cytokines and STAT3 in tumor and healthy renal tissue from 16 patients with histologically verified ccRCC. We have detected significantly higher levels of G-CSF, IL-6, CXCL10, CCL3, and CCL4 in tumor tissue than in their healthy counterparts. There were significant differences in the levels of IL-1ß and PDGF-BB between tumors of different nuclear grades (NG). Intratumoral IL-12p70 and IL-15 showed a significant positive correlation with intratumoral STAT3. The concentration of STAT3 in tumors was significantly lower than in the kidney. An increase in tumor STAT3 levels was associated with an increase in the pathological stage of the disease (TNM), but not with NG. The results of our study confirm the significant role of various cytokines and STAT3 in the pathogenesis of ccRCC and indicate their clinical relevance.

Analysis of HIF-1α expression and genetic polymorphisms in human clear cell renal cell carcinoma.

Introduction: Clear cell renal cell carcinoma (ccRCC) is mostly diagnosed incidentally and has relatively high recurrence rates. Alterations in VHL/HIF and mTOR pathways are commonly present in ccRCC. The present study attempted to identify potential diagnostic markers at the biochemical and molecular level. Methods: In total, 54 subjects (36 patients with ccRCC and 18 cancer-free controls) were enrolled. ELISA was used to measure the levels of HIF-1α in the tumor and healthy kidney tissue. The association between five selected SNPs (rs779805, rs11549465, rs2057482, rs2295080 and rs701848) located in genes of pathologically relevant pathways (VHL/HIF and mTOR) and the risk of ccRCC in the Slovak cohort was studied using real-time PCR. Results: Significant differences in HIF-1α tissue levels were observed between the tumor and healthy kidney tissue (p < 0.001). In the majority (69%) of cases, the levels of HIF-1α were higher in the kidney than in the tumor. Furthermore, the concentration of HIF-1α in the tumor showed a significant positive correlation with CCL3 and IL-1ß (p (R2) 0.007 (0.47); p (R2) 0.011 (0.38). No relationship between intratumoral levels of HIF-1α and clinical tumor characteristics was observed. Rs11549465, rs2057482 in the HIF1A gene did not correlate with the expression of HIF-1α either in the tumor or in the normal kidney. None of the selected SNPs has influenced the susceptibility to ccRCC. Conclusion: More research is neccesary to elucidate the role of HIF-1α in the pathogenesis of ccRCC and the association between selected SNPs and susceptibility to this cancer.

Cytokines in Renal Cell Carcinoma: A Step Towards Earlier Detection and Targeted Therapy.

Symptoms of renal cell carcinoma (RCC) have typically late onset and correlate with its advanced stage. No biomarkers of RCC are currently available. The present study analyzed the immuno-biochemical profile of RCC by measuring the levels of cytokines engaged in RCC pathophysiology. Cytokines were examined by capture sandwich immunoassays in tumor tissue and urine. Specimens of cancer and nearby healthy kidney tissues were obtained during nephrectomy from 60 RCC patients. The urine was obtained from both patients and healthy subjects. The findings in RCC tumor tissue compared to healthy renal tissues were following: (i) increases in interleukin-15 (IL-15), vascular endothelial growth factor (VEGF), interferon gamma-induced protein-10 (IP-10), macrophage inflammatory protein-1ß (MIP-1ß), monocyte chemoattractant protein-1 (MCP-1), and eotaxin, with VEGF, IP-10, and MIP-1ß significantly associated with the histologic tumor nuclear grading (NG); (ii) increases in platelet-derived growth factor (PDGF), IL-15, MIP-1ß, eotaxin, and MCP-1 in urine, with significant associations noticed between cytokines and disease stages for eotaxin and MCP-1; and (iii) decreases in PDGF, IL-15, MCP-1, VEGF, MIP-1ß, and eotaxin in urine from six patients on the third day after nephrectomy. We conclude that cytokines may play a critical role in the local pathogenesis of RCC, which opens the way for potential targeting of these molecules in novel therapies and their use as biomarkers for early noninvasive detection of RCC.

Tissue distribution of ethanol after intraprostatic injection using a porous needle.

Purpose: To develop a safe and precise method for intraprostatic injection, and to establish correlation between the volume of ethanol injectate and the volume of subsequent infiltrated prostate tissue. Materials and methods: We performed intraprostatic injection of 96% ethanol using a needle which has a segment of its wall made of capillary membrane with hundreds of pores in an acute and chronic canine experiment, in heart-beating cadaveric organ donors, and in a xenograft model of human prostate cancer. Whole mount tissue sections were used for three-dimensional reconstruction of the necrotic lesions and calculation of their volumes. Results: The ethanol injection resulted in oval shaped lesions of well-delineated coagulative necrosis. In both healthy human and canine prostates, the prostatic pseudocapsule and neurovascular bundle remained intact without evidence of disruption. There was a linear correlation between administered volume of ethanol and the volume of necrotic lesion. Regression analysis showed strong correlation in the acute canine experiments and in experiments performed on xenografts of human prostate cancer. A formula was calculated for each experiment to estimate the relationship between the injected volume and the volume of infiltrated prostate tissue area. Conclusions: Intraprostatic injection using a porous needle allows for effective and predictable tissue distribution of the injectate in the prostate. Through varying the volume of the agent injected and use of needles with a different length of the porous segment, the volume of infiltrated tissue could be adjusted allowing for targeted focal treatment.

Impact of innovative pelvic floor muscle training on quality-adjusted life years (QALYs) in women with stress urinary incontinence treated by duloxetine.

OBJECTIVES: The aim of this study was to measure the impact of innovative pelvic floor muscle training (iPFMT) on Quality-Adjusted Life Years (QALYs) in women with stress urinary incontinence (SUI) treated by duloxetine. STUDY DESIGN: This analysis is part of the DULOXING study conducted between February 2019 and 2020. The control group received oral duloxetine treatment (40 mg BID), and the experimental group received oral duloxetine treatment (40 mg BID) and iPFMT with lumbopelvic stabilization. SUI was analysed at baseline and in the final period according to the International Consultation on Incontinence Questionnaire-Urinary Incontinence Short Form (ICIQ-UI SF). The QALYs gained were calculated by multiplying life expectancy (LE) by a weighting factor (QALYs = LE * WF). RESULTS: The study included 158 women, of whom 129 were fully analysed (81.6%). The mean life expectancy was 26.3 ± 11.8 years for the control group and 29.0 ± 11.7 years for the experimental group. The mean baseline ICIQ-UI SF scores were 15.2 ± 1.7 vs 15.1 ± 1.5, and the final ICIQ-UI SF scores were 9.8 ± 4.2 vs 8.3 ± 3.8, in the control vs the experimental group, respectively (p < 0.05). The mean baseline WF was 0.27 ± 0.08 vs 0.28 ± 0.07, and the final WF was 0.53 ± 0.20 vs 0.60 ± 0.18, in the control vs the experimental group, respectively (p < 0.05). Before treatment, the number of QALYs during life expectancy in the control vs the experimental group was 7.53 ± 4.24 vs 8.30 ± 4.01. The number of QALYs during life expectancy in control vs the experimental group increased following treatment: 15.03 ± 7.63 vs 17.90 ± 7.86 (p < 0.05). CONCLUSIONS: Combination treatment with duloxetine and iPFMT statistically significantly increased the number of QALYs and reduced the degree of urinary incontinence in women with stress urinary incontinence.

Is health-related quality of life of patients after single-use flexible ureteroscopy superior to extracorporeal shock wave lithotripsy? A randomised prospective study.

The aims of the study were to compare the change in the Wisconsin Stone Quality of Life (WISQOL) score in patients who underwent retrograde intrarenal surgery (RIRS) single-use ureteroscope or extracorporeal shock wave lithotripsy (ESWL) with a calculation of quality-adjusted life-years (QALYs). 158 patients treated with urinary stone disease were randomly divided into 80 patients in the validation and 78 patients in the intervention arm. Patients in the intervention arm were randomly divided into the RIRS or the ESWL group. Linguistic validation of the WISQOL into the Slovak language was performed using a standardised multistep process. Discriminant validity was assessed by comparing stone-forming patients to an additional 34 healthy individuals. Patients were asked to fill in the WISQOL before and in the 24th week after the intervention. The QALYs were calculated by the formula QALY = weight factor (WF) x time period after intervention. The Cronbach's α of the WISQOL was 0.94, the Pearson's coefficient for test-retest reliability was 0.91, and the discriminant validity confirmed a higher score for healthy individuals (p < 0.001). The median WISQOL score changed from 45.5 to 95.5 vs. 33.9 to 87.1 in the RIRS and ESWL groups, respectively (p < 0.001). Patients from the RIRS group had a good possibility of reaching 19.727 QALYs gained during life expectancy compared to 15.780 for the ESWL group (p < 0.001). RIRS single-use ureteroscope is significantly superior to ESWL in reaching more QALYs gained during life expectancy. The WISQOL Slovak version is valid, reliable and strictly specific for stone-forming patients.

The Relationship between Overweight and Overactive Bladder Symptoms.

BACKGROUND: Several authors have investigated the relationship between obesity - assessed only by body mass index (BMI) - and overactive bladder (OAB) symptoms. OBJECTIVES: The objective of this study was to determine the relationship between body fat percentage (BFP) and the severity of OAB symptoms with an impact on quality of life. DESIGN: Cross-sectional study. The sample consisted of 206 overweight women; they were university students (BMI = 25.8 ± 3.0) with an average age of 30.6 ± 2.4 years. Body composition analysis was used, including assessment of BFP, visceral fat area (cm2/level), and other parameters. OAB symptoms were evaluated with an OAB questionnaire (OAB-q), voiding diary, and quality of life scale (I-QoL). RESULTS: Ninety women had a BFP >32% and 116 had a BFP <32%. The voiding diary and OAB-q confirmed significant differences in 24-h daytime and nighttime frequency of voiding and average urine volume during the day. The OAB-q symptom score was lower in the group with BFP <32%. I-QoL recorded significantly worse parameters in the group with BFP >32% (p < 0.01). The Patient Perception of Intensity of Urgency Scale significantly correlated with BFP (r = 0.466, p < 0.001). Women with a BFP above 32% had a 1.95 times greater chance of developing OAB (odds ratio = 1.95, 95% CI = 1.09-3.52, p < 0.02). CONCLUSION: Young women with a BFP >32% were 95% more likely to have OAB than other young women with a BPF <32%.

Involvement of calcium regulating ion channels in contractility of human isolated urinary bladder.

This study specified the role of several key calcium-operating ion channels in contraction/relaxation of human detrusor muscle as possible target for overactive bladder (OAB) treatment. Detrusor samples, obtained from 18 males (average age 61.5 ± 5.9 years), were investigated by organ tissue bath method with following agents: diltiazem for L-type voltage-gated calcium channels; 3-fluropyridine-4-carboxylic acid (FPCA) for Orai-STIM channels; SKF 96365-hydrochloride for transient receptor potential (TRP) channels, T-type channels and Orai-STIM channels; 2- aminoethoxydiphenyl borate (2-APB) for inositol-triphosphate receptors (IP3Rs) and Orai-STIM channels. Oxybutynin and mirabegron were tested under the same conditions as controls. Mirabegron, 2-APB and FPCA exhibited the best suppressive effect on carbachol-induced detrusor contractility. As expressed by area under the contractile curve (AUCC), 2-APB, FPCA and mirabegron have similar AUCC: 1.79, 1.73, 1.73. The highest AUCC was 3.64 for diltiazem+SKF, followed by 3.21 for diltiazem, 3.16 for SKF and 2.94 for oxybutynin. The lowest median amplitude and contraction variability is for 2-APB followed by mirabegron and FPCA. There were significant differences between: 2-APB/FPCA vs.: ditiazem, diltiazem+SKF and SKF. Summary of results suggested the principal role of IP3Rs, Orai-STIM coupling and large-conductance calcium-activated potassium channels in detrusor contraction and pointed on Orai-STIM channels as possible targets for OAB pharmacotherapy.

Impact of clean intermittent catheterization on quality adjusted life years (QALYs) in spinal cord injury patients with neurogenic urinary incontinence.

AIMS: The impact of clean intermittent catheterization (CIC) on quality adjusted life years (QALYs) gained in adults' spinal cord injury population with neurogenic urinary incontinence (UI). METHODS: Patients were recruited from the national registry January-June 2014. The inclusion criteria were adults, neurogenic UI due to spinal cord injury (SCI), use of collection devices and CIC for more than 6 months. The exclusion criteria were inability to perform CIC, cancer of the lower urinary tract and fistulas formation. Measurement tools were the International Consultation on Incontinence Questionnaire-Urinary Incontinence Short Form (ICIQ-UI SF) and an estimation of life expectancy by the national registry. The calculation of the weighting factor (WF) was obtained by linear transformation of the ICIQ-UI SF total score. A score was transformed to the range from 0 (worst impact) to 1 (no impact). The QALYs was calculated as the weighting factor × life expectancy in years. RESULTS: A total of 229/365 patients were involved in this study (63%). Patients before CIC reached an ICIQ mean score of 14.83, WF of 0.29, and QALYs of 9.02 during life expectancy. After 6 months of follow-up using CIC, ICIQ reached 9.12, WF 0.57 and QALYs 17.45. The number of QALYs increased by 93.5% and UI evaluated with the ICIQ-UI SF decreased by 38.5% (P < 0.01). CONCLUSIONS: The CIC of the urinary bladder statistically significantly increased the number of QALYs and reduced the degree of UI in SCI patients.

Orai1 protein expression and the role of calcium release-activated calcium channels in the contraction of human term-pregnant and non-pregnant myometrium.

AIM: This experimental in vitro study examined differences in the expression and activity of calcium release-activated calcium (CRAC) channels of human term-pregnant and non-pregnant myometrium. MATERIAL AND METHODS: The tissue samples were obtained from term-pregnant myometrium in labor of women undergoing cesarean section and from non-pregnant myometrium of women undergoing total hysterectomy due to uterine myoma. The expression of Orai1 protein, a pore-forming subunit of CRAC channels, in human myometrium was examined using immunohistochemistry. CRAC channel involvement in the amplitude and frequency of myometrial contractions was evaluated in vitro using a tissue bath method with a CRAC ion channel blocker 3-fluropyridine-4-carboxylic acid (FPCA). RESULTS: Decreased Orai1 expression was observed in human term-pregnant laboring myometrium compared with non-pregnant myometrium. However, the initial oxytocin-induced contraction of myometrium was significantly suppressed at different doses of FPCA in both non-pregnant human isolated myometrium and non-pregnant myometrium. The frequency of contractions was the most significantly reduced at the lowest dose of FPCA in non-pregnant myometrium and remained suppressed at all doses of FPCA in term-pregnant myometrium. Salbutamol was shown as more effective in suppression of amplitude in term-pregnant isolated myometrium. CONCLUSION: Our results provide the first information about the changes in the Orai1 protein expression and activity of human myometrial CRAC channels in term-pregnant laboring myometrium.

Disability-adjusted life years (DALYs) in general population with pelvic organ prolapse: a study based on the prolapse quality-of-life questionnaire (P-QOL).

OBJECTIVE: To evaluate the impact of pelvic organ prolapse (POP) on quality of life in symptomatic and asymptomatic women. STUDY DESIGN: This is a cross-sectional study in a general population of adult women. Symptomatic and asymptomatic women with POP were studied. Symptomatic women were defined as those who reported the presence/feeling of lump/bulge/pressure in the vagina and on vaginal examination had the sign of POP defined as the leading edge of the vagina wall/cervix presenting at or beyond the hymeneal remnants. All women were asked to self-complete the Prolapse Quality of life questionnaire (P-QOL) to estimate the disability-adjusted life-years (DALYs) lost due to symptomatic POP. DALYs for a symptomatic POP were calculated as the sum of the years of life lost due to premature mortality (YLL) in the population and the years lost due to disability (YLD). To determinate the optimal threshold of each P-QOL domain in relationship to the symptomatic POP population a receiver operating curve (ROC) and area under curve (AUC) analysis were used. RESULTS: A total of 785 were recruited. Only 539 (68.7%) consented to participate in the study. The mean age was 47 years (range 18-82 years). The total scores for each of the P-QOL domains were found to be significantly higher in symptomatic prolapse subjects compared to asymptomatic subjects (p<0.05). Estimated DALY lost per year per 1000 women obtained were 217.0 vs 324.8, lost years of 14.5 vs 10.3 in 50 vs 60-year old women. CONCLUSION: In this group of women with POP the quality of life is severely affected. The DALYs revealed lost years due to symptomatic POP.

Gender-specific external barriers to seeking care for urinary incontinence.

BACKGROUND: Barriers to seeking care for urinary incontinence are specific, objective, external conditions that prevent incontinence sufferers from seeking treatment. The aim of this study was to compare barriers, gender, and health care disparities in incontinence sufferers. METHODS: Incontinent patients were recruited into a questionnaire-based cross-sectional study. The 14-item Barriers to Incontinence Care Seeking Questionnaire (BICS-Q) and the three-item International Consultation on Incontinence Questionnaire-Urinary Incontinence Short Form (ICIQ-UI SF) were used to evaluate barriers to seeking health care for urinary incontinence. RESULTS: The representative sample (n = 1014) finally included 567 adults eligible to participate in this study (response rate 55.9%). Of the 147 incontinent males, 93 (63.3%) did not seek care, and of the 420 incontinent females, 282 (67.1%) did not seek care. Untreated males had significantly higher BICS-Q scores than other patients. Risk factors for barriers were obesity (odds ratio 2.13 for females versus 0.83 for males), stress urinary incontinence (1.57 versus 9.38, respectively), and urgency urinary incontinence (2.40 versus 1.75). CONCLUSION: The barriers to seeking care for urinary incontinence seem to be gender-specific. Obese females with urgency urinary incontinence and males with stress urinary incontinence were least likely to seek treatment.

Gene polymorphisms in bladder cancer.

In Europe, cancer of the bladder is the fourth most common cancer among men, accounting for 7% of total cancers. In the USA, bladder cancer is the fifth most common cancer in men and seventh in women. This disease is three times more common in men than in women. Several risk factors, such as cigarette smoking and occupational chemical exposure, contribute to bladder cancer development. The balance between activation and detoxification of carcinogens affects the amount of DNA damage that accumulates in cells. The entire process leading to DNA damage and subsequent repair of the damage involves a host of enzymes, many of which are polymorphic. Polymorphisms in metabolic enzyme genes and repair genes may cause alterations in protein product functions that can finally lead to genomic instability and carcinogenesis. In this article, we review the polymorphisms in a number of genes that have been found to be the modulators of bladder cancer risk. Improved understanding of the molecular biology of urothelial malignancies is helping to more clearly define the role of new prognostic indices and multidisciplinary treatment for this disease.