Synergic effects of cancer stem cells markers, CD44 and embryonic stem cell transcription factor Nanog, on bladder cancer prognosis.

Background: Therapy that targets cancer stem cells has the potential to eradicate cancer and prevent tumour recurrence. Therefore, we hypothesized the combined prognostic significance of stem cell markers CD44 (prevalent in basal layer of urothelial carcinoma) and Nanog (embryonic stem cell transcription factor) in bladder cancer.Material and Methods: CD44 and Nanog expression were determined by immunohistochemistry in 112 bladder cancer cases of which 79 were non-muscle invasive and 33 muscle invasive.Results: A significant correlation was found between CD44 and Nanog expression (r = 0.41, p < 0.001). The bladder cancer patients with high CD44 and Nanog expression had poor recurrence-free survival and poor overall survival (all p < 0.01). Multivariate Cox regression analysis identified lymph node positivity (hazard ratio; HR 3.81, 95% confidence interval; CI 1.66-8.75), CD44 (HR/95%CI 7.03 [3.04-16.22]) and Nanog (HR/95%CI 2.89 [1.23-6.77]) as independent prognostic biomarkers for recurrence-free survival, whilst a combined index of CD44 and Nanog expression (high expression group; HR/95%CI 25.45 [6.71-96.50] for recurrence-free survival) and lymph node positivity (HR/95%CI 3.68 [1.63-8.33] for recurrence-free survival) were independent prognostic biomarkers for recurrence-free survival and overall survival (all p < 0.001).Conclusions: A combined index of CD44 and Nanog expression is a promising prognostic predictor of recurrence-free survival and overall survival in bladder cancer. It may help identification of patients who will benefit from intensive treatment.

Functional outcomes of clinically high-risk prostate cancer patients treated with robot-assisted radical prostatectomy: a multi-institutional analysis.

BACKGROUND: To ascertain 3-year urinary continence (UC) and sexual function (SF) recovery following robot-assisted radical prostatectomy (RARP) for clinically high-risk prostate cancer (PCa). METHODS: Retrospective analyses of a prospectively maintained database for 769 patients with D'Amico high-risk PCa undergoing RARP at two tertiary care centers in the United States and Europe between 2001 and 2014. The association between time since RARP and recovery of UC (defined as 0 pad/one safety liner per day) and SF (defined as sexual health inventory for men (SHIM) score ⩾17) was tested in separate preoperative and post-operative Cox-proportional hazards regression models. Sensitivity analyses were conducted using continence 0 pad per day and erection sufficient for intercourse as end points for UC and SF recovery, respectively. RESULTS: Mean age of the cohort was 62.3 years, and 62.1% harbored ⩾PT3a disease. Nerve sparing (unilateral or bilateral) RARP was performed in 87.7% of patients. Kaplan-Meier estimates of UC recovery at 12, 24 and 36 months after surgery was 85.2%, 89.1% and 91.2%, respectively, while 33.8, 52.3 and 69.0% of preoperatively potent men (preoperative SHIM ⩾17; n=548; 71.3%) recovered SF. Similar results were noted in sensitivity analyses. Patient age and year of surgery were associated with UC and SF recovery; additionally, preoperative SHIM score, degree of nerve sparing, pT3b-T4 disease and surgical margins were associated with SF recovery over the period of observation. CONCLUSIONS: Patients with D'Amico high-risk PCa treated with RARP may continue to recover UC and SF beyond 12 months of surgery and show promising outcomes at 3-year follow-up. Appropriate patient selection and counseling may aid in setting realistic expectations for functional recovery post RARP.

Expression and clinical significance of Centrosomal protein 55 (CEP55) in human urinary bladder transitional cell carcinoma.

Bladder cancer (BC) is one among the most common and lethal urothelial malignancies worldwide. The expression of cancer-testis (CT) antigens in some tumours and restricted expression among normal tissues make CT antigens as attractive vaccine targets. In this context, we evaluated Centrosomal protein 55 kDa (CEP55), which is specifically expressed in normal human testis and various malignancies. Until the expression pattern of CEP55 in transitional cell carcinoma (TCC) of human urinary bladder and its clinical significance are not known. The aim of the present study is to evaluate mRNA/protein expression of CEP55 in TCCs of urinary bladder and correlate its expression with the clinicopathological characteristics of BC patients. In this study, the methods of quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) were used to investigate mRNA/protein expression of CEP55 in TCC. Independent Student's t test, ANOVA and Chi-square (χ(2)) were used to analyze the data statistically. We observed CEP55 mRNA overexpression in testis and 48.7% of BC patients. Relative mean fold expression of CEP55 mRNA was found to be significantly (p<0.01) higher in muscle-invasive bladder cancer (MIBC) as compared to non-muscle-invasive bladder cancer (NMIBC) patients (7.88±3.88 vs. 4.75±2.30, p=0.01). CEP55 protein expression was evaluated using IHC and cytoplasmic staining pattern was recorded in formalin fixed, paraffin-embedded (FFPE) bladder tumour tissues. No significant difference was observed in protein expression of CEP55 between the two groups (NMIBC and MIBC patients) (72.2% vs. 69.0%, p=0.774). No significant protein expression of CEP55 was observed among adjacent noncancerous tissues (ANCTs) and benign prostatic hyperplasia (BPH) used as control. Our study results suggest that CEP55 mRNA/protein expression was observed is specific to TCC of human urinary bladder and might be used as a diagnostic biomarker and vaccine target in development of BC specific immunotherapy.

Appraisal of diagnostic ability of UCA1 as a biomarker of carcinoma of the urinary bladder.

Initial diagnosis of carcinoma of the urinary bladder remains to be a challenge. Urine cytology, as an adjunct to cystoscopy, is less sensitive for low-grade tumors. Urothelial cancer associated 1 (UCA1) is a novel non-coding RNA gene, which plays a pivotal role in bladder cancer progression. Our aim is to investigate the significance of urinary UCA1 for the non-invasive diagnosis of transitional cell carcinoma (TCC) of the urinary bladder. We examined UCA1 expression in a bladder cancer cell line (T24) and in urine of 28 healthy individuals, 46 patients of non-malignant disorders, and 117 cases (69 primary and 48 recurrent cases) of histologically proven TCC prior to transurethral resection by using real-time PCR and compared it with voided urinary cytology. UCA1 expression was found in T24 cell line and also found to be significantly higher in the cancer group as compared to the controls (p<0.001). UCA1 messenger RNA (mRNA) expression showed a significant (p<0.05) association with stage and grade (p<0.05). UCA1 showed a sensitivity of 79.49% and a specificity of 79.73% (p<0.001), whereas urine cytology had a sensitivity of 66.67% and a specificity of 95.95% for TCC cases. Higher expression of UCA1 was associated with high grade (G2-G3, sensitivity=84.09%) (p<0.001). UCA1 mRNA expression did not significantly correlate with the patient's age, sex, and smoking habit (p>0.05). UCA1 can be used as a non-invasive diagnostic biomarker for TCC bladder as an adjunct to cytology in the early diagnosis of primary urinary bladder cancer.

Evaluation of urinary XIAP as a diagnostic biomarker of carcinoma of urinary bladder.

Early diagnosis of carcinoma of the bladder remains a challenge. Urine cytology, as an adjunct to cystoscopy, is less sensitive for low-grade tumors. X-linked inhibitor of apoptosis (XIAP), a negative regulator of apoptotic stimuli and a member of apoptosis family, is frequently activated in bladder carcinoma. Our aim is to investigate the significance of urinary XIAP for the noninvasive diagnosis of transitional cell carcinoma (TCC) of the urinary bladder. We examined urinary XIAP expression in a bladder cancer cell line (T24) and in urine of 28 healthy individuals, 46 patients of nonmalignant disorders, and 117 cases (69 primary and 48 recurrent cases) of histologically proven TCC prior to transurethral resection, by using real-time PCR, and compared it with voided urinary cytology (VUC). XIAP expression was found in T24 cell line and also was found to be significantly higher in the cancer group as compared to the controls (p < 0.001). XIAP messenger RNA (mRNA) expression showed a significant (p < 0.05) association with stage and grade (p < 0.05). XIAP shows the sensitivity of 82.91 % and specificity of 78.38 % (p < 0.001), whereas urine cytology had sensitivity of 66.67 % and specificity of 95.95 % for TCC cases. The combination of XIAP and VUC had better sensitivity (98.2 %) and specificity (92.6 %) than they showed individually (p < 0.001). XIAP mRNA expression did not significantly correlated with the patient's age, sex, and smoking (p > 0.05). Urinary XIAP can be used as a noninvasive diagnostic biomarker for bladder TCC in adjunct to cytology mainly for low-grade non-muscle-invasive tumors.

Expression of PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK) in human urinary bladder transitional cell carcinoma.

The objective of this study was to evaluate the expression pattern of PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK) and its clinical significance in human bladder cancer (BC). We detected PBK/TOPK mRNA overexpression in BC and human normal testis tissues using RT-PCR. Using qRT-PCR revealed a higher expression of PBK/TOPK in BC tissues than their adjacent noncancerous tissues (ANCTs) (p<0.0001). Cytoplasmic expression of PBK/TOPK protein was found to be positive in 64.6% (42 of 65) BC patients. Expression of PBK/TOPK protein was found to be significantly higher in muscle-invasive bladder cancer (MIBC) than in non-muscle-invasive bladder cancer (NMIBC) (86.1% vs. 37.9%, p<0.001). The immunohistochemical (IHC) expression of PBK/TOPK was found to be significantly (p<0.001) associated with the stage of disease. Study findings suggest that the PBK/TOPK mRNA/protein expression is specific to human BC and might be used as a novel target for development of cancer immunotherapy and diagnostic biomarker.

Selective estrogen receptor modulators for BPH: new factors on the ground.

As the current management of BPH/lower urinary tract symptoms by traditionally involved pharmacological agents such as 5alpha-reductase inhibitors and α1-adrenoceptor antagonists is suboptimal, there is definite need of new therapeutic strategies. There is ample evidence in literature that suggests the role of estrogens in BPH development and management through the different tissue and cell-specific receptors. This article reviews the beneficial actions of selective estrogen receptor modulator (SERM) and ERß-selective ligands, which have been demonstrated through in vitro studies using human prostate cell lines and in vivo animal studies. SERMs have anti-proliferative, anti-inflammatory and pro-apoptotic mechanisms in BPH, and also act by inhibiting various growth factors, and thus represent a unique and novel approach in BPH management directed at estrogen receptors or estrogen metabolism.

Plasma vascular endothelial growth factors A and C in patients undergoing prostatic biopsy and TURP for suspected prostatic neoplasia.

BACKGROUND: Formation of new blood vessels is necessary for the development and spread of neoplasms more than 1 mm3 in volume, angiogenesis being responsible for formation of new from pre-existing blood vessels. Vascular endothelial growth factor (VEGF) is pivotal and the best studied angiogenic factor in all human cancers. Therefore we designed this study to investigate the role of VEGF-A and VEGF-C in prostate cancer in comparison with BPH controls in a north Indian population. METHODS: In this case-control study a total of 100 subjects were included on the basis of confirmed histopathological reports, out of which 50 were prostate cancer patients and the other 50 were BPH patients with PSA levels >2 ng/ml and abnormal digital rectal examination (DRE) findings during September 2009 to August 2011 from the Department of Urology, KGMU, Lucknow, India. Plasma levels of VEGF were determined using quantitative immunoassay (ELISA- enzyme linked immunosorbent assay). Statistical analysis was carried out using SPSS 15.0 version. RESULTS: The mean age of prostate cancer (67.6±5.72) patients was significantly higher (p=0.005) than BPH (63.6±7.92) patients. Expression of VEGF-A was not significantly higher in disease stage C1 than D1 or D2 and A or B (p=0.13) while the level of VEGF-A was significantly higher (p=0.04) in prostate cancer as compared to BPH subjects (PCa=13.0 pg/ml, BPH=6.8 pg/ml). Levels of VEGF-C were similar in both groups (PCa=832.6 pg/ml, BPH=823.7 pg/ml). In ROC curve, the area under curve (AUC) was 0.70 (95%CI: 0.60-0.80) and the cut-off value for which a higher proportion of patients was correctly classified (20%) was 26.0 pg/mL. CONCLUSION: Although VEGF-A is increased in cancer prostate patients a statistically significant correlation could not be established in this study. VEGF-C was not found to be a useful biomarker.

A novel midline scroto-perineal approach facilitating innervation preserving sphincteroplasty and radical corporal detachment for reconstruction of exstrophy-epispadias.

OBJECTIVE: To report a novel surgical approach for single-stage repair of exstrophy-epispadias, with the specific aim of innervation, preserving sphincteroplasty corporal detachment for penile lengthening and ischio-pubic ramotomy for Linia-alba approximation. MATERIAL AND METHODS: Twenty-five classic exstrophy with compliant bladder plate and 10 incontinent epispadias patients were selected. Preoperative magnetic resonance angiogram of urogenital diaphragm showed triangular space between ischio-cavernosus, bulbo-spongiosus, and transverse-perinei muscles containing sphincteric branch of perineal artery indicating the course of sphincteric nerve. Bladder plate was mobilized. Through a midline scrotal septal and transverse incision along the base of urogenital triangle, the urogenital diaphragm was exposed. Corpora were separated from the urethral plate while preserving the glanular continuity and innervation to striated urethral sphincter, using muscle stimulator and nerve integrity monitor. In the subperiosteal plane along the ischio-pubic rami, the corpora were detached. Repair included ureteric reimplantation; anatomic reconstruction of bladder, bladder neck, urethra, and striated sphincter; corporo-glanuloplasty; ischio-pubic ramotomy; and abdominal closure. Assessment included surgical problems, cosmetic satisfaction, erectile function, continence, and upper tract status at 2-year follow-up. RESULT: There was no corporal loss. Postoperative complications included 4 perineal suture line infections, 11 peno-pubic fistula, and 1 adhesive intestinal obstruction. Erectile function was good in 33 patients. Penile length was gratifying in 25. Of 28 patients, 20 (71.4%) had dry interval of two hours. Dimercaptosuccinic acid study demonstrated upper tract scarring in 2 patients. CONCLUSION: This approach facilitates innervation preserving sphincteroplasty and precise restoration of anatomy to near normal without operative accidents because of wide exposure gained, improving the functional and cosmetic results.

Is central obesity, hyperinsulinemia and dyslipidemia associated with high-grade prostate cancer? A descriptive cross-sectional study.

AIMS: The association of central obesity, hyperinsulinemia, and dyslipidemia with higher grade advanced prostate cancer as determined by Gleason grading is not well understood. We evaluated the effect of central obesity waist hip ratio (WHR ≥ 0.9) and biochemical parameters associated with central obesity on Gleason grading in North Indian patients of prostate cancer presenting at advanced stages. MATERIALS AND METHODS: A cross-sectional study was conducted among 50 nondiabetic patients having clinical stages III and IV prostate cancer. Gleason grading on core biopsy samples by histopathology was done and patients were divided in two groups-group1, Gleason score ≥8; group 2, Gleason score <8. WHR along with serum levels of prostate-specific antigen (PSA), testosterone, insulin, and lipid profile was done in each patient. RESULTS: The two groups are similar in Age (67.54 years); range (50-80 years). Group 1 men had statistically higher mean WHR (0.96 vs 0.90; P ≤ 0.001), higher mean triglyceride level (201.34 vs 150.52 mg/dL; P=0.0006), higher mean very low-density lipoprotein (VLDL) (40.27 vs 30.10 mg/dL; P =0.0006), higher mean insulin (19.49 vs 15.04 µIU/mL; P = 0.0024), and lower mean high-density lipoprotein (HDL) levels (32.39 vs 36.82 mg/dL; P = 0.034) than men in group 2. Serum levels of cholesterol, LDL, and testosterone did not show statistically significant differences between the two groups. CONCLUSIONS: This pilot study involving small number of patients indicates that central obesity, dyslipidemia, and hyperinsulinemia could be associated with high-grade prostate cancer.

A cross-sectional pilot study to determine the prevalence of testosterone deficiency syndrome in working population of Indian men.

AIM: To determine the prevalence of testosterone deficiency syndrome (TDS) in healthy Indian men employed in a hospital aged above 40 years. MATERIALS AND METHODS: A general medical health check-up camp was organized for all male employees above 40 years age working in surgical departments. After clinical history and systemic inquiry, subjects were requested to fill the St. Louis University's ADAM Questionnaire based on which the total and free-serum testosterone estimation was then done. RESULTS: One hundred fifty seven healthy volunteers enrolled for the study (mean age 53.1 years; range 40-60). The androgen decline in the aging male (ADAM) Questionnaire detected 106 men (67.5%) to be symptomatic for TDS. Serum testosterone estimation in these subjects revealed 41/106 to have low free-serum testosterone levels and 32/106 to have low total-serum testosterone. In 11 and 6 cases, respectively, the serum free- and total-testosterone levels were found to be low although the subjects were asymptomatic for TDS. CONCLUSIONS: The prevalence of symptomatic biochemical hypogonadism was 26.1%. The higher prevalence of symptoms alone of TDS was unusual. It could be because of the nature of the questionnaire. Free-serum testosterone may be a better single test to diagnose symptomatic hypogonadism than total-serum testosterone.

Heterotopic bone formation in renal cell carcinoma: a diagnostic challenge.

Formation of bone in cases of renal cell carcinoma is a rare finding and only a couple of case reports from Japan and one from India are mentioned in the literature. Calcification inside renal mass has been reported earlier but the prognostic implications have not been clearly elucidated. We report a case which showed heterotopic bone formation (ossification) inside the renal mass and was managed by radical nephrectomy. The histopathology showed clear cell renal carcinoma with multiple centers of ossification in the region of calcification suggesting bone formation. In this case report we discuss bone morphogenetic proteins which have been implicated as a prognostic and causative factor, highlight the difficulties in distinguishing between calcification and bone formation on the basis of radiological investigations and mention the geographic implications of this rare phenomenon which has not been described earlier.

Harvesting buccal mucosa graft under local infiltration analgesia--mitigating need for general anesthesia.

For buccal mucosal graft urethroplasty, nasal or oral endotracheal intubation anesthesia is used for harvesting the graft from the oral cavity. A technique of graft harvesting under local anesthesia using 2% lidocaine solution with adrenaline (1:200,000) is described. This method requires a cooperative patient but saves the morbidity of general anesthesia.

Options in the management of tuberculous ureteric stricture.

Ureteric stricture is a feared manifestation of genitourinary tuberculosis (TB) with the commonest site being the lower ureter. The purpose of this review is to discuss the management options for this condition. Literature search was done using PubMed and all articles on TB and ureteric stricture were reviewed published between 1990 till September 2007. The exact site and length of stricture must be defined with radioimaging (intravenous urography, retrograde, or antegrade pyelography) and renal function be quantified. The treatment of stricture mostly requires some kind of intervention after a brief period of antituberculous medicines with or without steroids. For uncomplicated/simple strictures (short segment, passable, with renal function >25%, good bladder capacity) endourologic option should be used which usually means double-J stenting with or without balloon dilatation. For complicated/complex strictures (long segment, dense fibrosis, with renal function <20%, small bladder capacity) regular surgical options should be considered which usually means ureteroureterostomy or ureteropyelostomy for upper ureteric strictures, intubated ureterostomy, or transureteroureterostomy for midureteric strictures, psoas hitch/Boari flap for lower ureteric strictures or ileal ureter/autotransplantation for whole length/multiple strictures.

Guidewire-assisted infundibular tilt: a technique for access into another calix during percutaneous nephrolithotomy.

Sometimes, during percutaneous nephrolithotomy, a stone fragment migrates into a parallel calix, necessitating a separate puncture. We describe a simple technique for removing such fragments. This technique is especially useful for patients who have moderate to severe hydronephrosis where the intercaliceal parenchyma is thin.

Renal capsular block: a novel method for performing percutaneous nephrolithotomy under local anesthesia.

Most of the pain of percutaneous nephrolithotomy is caused by dilatation of the renal capsule and parenchymal tract. We evolved a technique to block renal capsular sensations by infiltration of lignocaine (lidocaine) at the site of renal entry, permitting the procedure to be done with local anesthesia.

Silver nitrate sclerotherapy for 'clinically significant' chyluria: a prospective evaluation of duration of therapy.

OBJECTIVES: To evaluate and compare the results of regimen A (3 instillations at 8-hourly intervals in 1 day) with the control regimen B (9 instillations at 8-hourly intervals in 3 days) of using 1% silver nitrate solution for renal pelvic instillation sclerotherapy in 'clinically significant' filarial chyluria. MATERIALS AND METHODS: Forty-seven patients with clinically significant chyluria attending on 2 different days our urology clinic were prospectively randomized between two groups; the study group received regimen A (n = 21) while the control group received regimen B (n = 26). The variables evaluated included visualization of pyelolymphatic fistulae on retrograde pyelography, hospital stay, outcome and morbidity of the two regimens. RESULTS: Patients in both groups were comparable for age and sex. The morbidity (fever, symptomatic UTI, hematuria) following regimen A was less than that of regimen B although not statistically significant. The average hospital stay was 3 days for regimen A and 5.5 days for regimen B (p = 0.001). The initial success rate was 80.95% in group A and 92.30% in group B (p = 0.47). The mean duration of follow-up was 15 months (range 9-18). There was no significant difference in recurrence between the two groups during follow-up (group A: 21.05% and group B: 22.72%; p = 0.98). CONCLUSIONS: Regimen A was as effective as regimen B. Regimen A had the advantages of having less morbidity and shorter duration of hospital stay. We recommend only a 3-instillation regimen in patients with clinically significant chyluria, particularly those who demonstrate pyelolymphatic fistulae on retrograde pyelography.