Endogenous BTG2 expression stimulates migration of bladder cancer cells and correlates with poor clinical prognosis for bladder cancer patients.

BACKGROUND: The B-cell translocation gene 2 (BTG2) is considered to act as a tumour-suppressor gene because of its antiproliferative and antimigratory activities. Higher levels of BTG2 expression in tumour cells have been linked to a better clinical outcome for several cancer entities. Here, we investigated the expression and function of BTG2 in bladder cancer. METHODS: The expression of BTG2 in bladder cancer cells was silenced by RNA interference. Cell motility was investigated by wound healing and Boyden chamber assays. The protein expression of BTG2 in bladder cancer was studied by immunohistochemistry. RESULTS: We observed that targeted suppression of BTG2 by RNA interference did not result in growth stimulation but led to a substantial inhibition of bladder cancer cell motility. Tissue microarray analyses of bladder cancer cystectomy specimens revealed that higher BTG2 expression levels within the tumours correlated strongly with a decreased cancer-specific survival for bladder cancer patients. CONCLUSION: These results indicate that endogenous BTG2 expression contributes to the migratory potential of bladder cancer cells. Moreover, high levels of BTG2 in bladder cancers are linked to decreased cancer-specific survival. These findings question the conception that BTG2 generally acts as a tumour suppressor and typically represents a favourable clinical marker for cancer patients.

Prediction of recurrence and progression in patients with T1G3 bladder cancer by gene expression of circulating tumor cells.

OBJECTIVE: To investigate the role of gene expression of circulating tumor cells (CTCs) as noninvasive prognostic markers in patients with high risk nonmuscle invasive bladder cancer. MATERIALS AND METHODS: We identified all patients with TIG3 urothelial bladder cancer (UBC) at our institution since 2016.The study included 100 patients with T1G3 UBC and 50 healthy volunteers. CTCs were isolated from blood using immunomagnetic separation and gene expression was performed using 10 bladder cancer associated genes, namely; KRAS, EPCAM, CD133, CD44, mTOR, SURVIVIN, AKT, PI3K, VEGF, and TP53. Gene expression of CTCs was correlated to time to first recurrence and time to progression using Kaplan-Meier curves. RESULTS: There was strong negative correlation between CTCs-positive patients and time to first recurrence and time to progression. Significant differences in expression levels of specific genes were observed that can predict recurrence and progression of T1G3 UBC. CONCLUSION: CTCs appear to be noninvasive methods of predicting disease recurrence and progression in patients with high- risk nonmuscle invasive bladder cancer; therefore, studying their molecular profiling may improve prediction of recurrence and progression. Further studies are invited for more in-depth investigation to consolidate our initial results.

Renal transplantation in children with abnormal lower urinary tract.

OBJECTIVE: This retrospective study reports the outcomes as well as pre- and posttransplant urologic treatments of renal transplantation for children with an abnormal lower urinary tract (LUT). METHODS: Between March 1981 and December 2001, 195 children (< or =18 years of age) received live-donor kidney transplants. The 15 recipients (14 boys and 1 girl, mean age 13.5 +/- 3 years) who had lower urinary tract disorders included posterior urethral valves (PUV) with valve bladder (n=12) and neuropathic bladders secondary to meningomyelocele (n=3). These children were evaluated by voiding cystourethrogram, cystourethroscopy, and cystometry. The children with PUV were maintained on clean intermittent catheterization (CIC) and a detrusor relaxant at least 3 months before transplantation. Augmentation ileocystoplasty or continent cutaneous diversion were used in three patients. The graft and patient survivals as well as complications in this cohort was compared with a group of children with normal LUT, who underwent renal transplantation during the same period. RESULTS: One child died in the early posttransplant period due to rupture of the external iliac artery. Follow-up ranged from 6 months to 16 years (mean=4.5 years). During the same period the graft and patient survival rates were comparable between the group of children with versus without abnormal LUT. Furthermore, mean serum creatinine and creatinine clearance values were also comparable. The group with an abnormal LUT showed a higher incidence of urinary fistula (3/14) and recurrent UTI and/or bacteremia (4/14). CONCLUSIONS: Renal transplantation is feasible with good results for children with abnormal LUT. Pre- and posttransplant urologic management is critical for a successful outcome. However these children display a high incidence of urologic and infectious complications.

Renal ischaemia/reperfusion injury: possible role of aquaporins.

Renal ischaemia/reperfusion (I/R) injury is a common problem that occurs when blood flow is interrupted to the kidney in case of kidney transplantation, aortic cross-clamping and shock with subsequent resuscitation. Renal I/R injury is a complex conditions which includes the onset of an inflammatory process, which is associated with impairment of concentrating ability of the kidney and impairment of solute transport. Characteristically, renal I/R injury is associated with marked reduction in the protein expression of renal aquaporins (AQPs) mainly (AQP1, AQP2 and AQP3), and solute transporters were observed in this condition and could account for the impaired urinary concentration that observed in this condition. Recently, many agents were tested for a possible protective effect against this insult such as erythropoietin (EPO), α-melanocyte-stimulating hormone (α-MSH) and α-lipoic acid which were proved to prevent downregulation of AQPs and solute transporters. The aim of this short review is to outline the potential pathophysiological role of AQPs in renal I/R injury and to put a spotlight on the modulation of renal functions impairment in renal ischaemia by new drugs that prevent downregulation of AQPs.

Serous-lined extramural ileal valve as a new continent cutaneous urinary outlet: an experimental study in dogs.

A new technique for the construction of a cutaneous continent catheterizable outlet suitable for urinary reservoirs is presented. The technique entails the creation of an ileal reservoir outlet by implantation of a tapered ileal segment into a serous-lined extramural tunnel. The procedure was carried out in eight experimental dogs, and the results were compared with a control group of five dogs for which a nontunnelled tapered ileal outlet draining a similar reservoir was constructed. Clinical, radiological and urodynamic evaluation provided evidence that this new outlet is easily catheterizable. Its continence mechanism is more reliable than that of a simple tapered outlet.

A novel uretero-ileal reimplantation technique: the serous lined extramural tunnel. A preliminary report.

A novel technique for an anti-refluxing uretero-ileal reimplantation entailing creation of 2 serous lined extramural tunnels in a detubularized ileal W-bladder is presented. The operation was done on 12 patients in whom an orthotopic bladder substitute was indicated. Mean followup was 18 months. Evidence indicated that this method could provide a nonobstructed unidirectional flow of urine in all of the examined renal units.

Functional results of orthotopic ileal neobladder with serous-lined extramural ureteral reimplantation: experience with 450 patients.

PURPOSE: We report functional results of the orthotopic ileal neobladder using a serous-lined extramural tunnel as an antireflux procedure. MATERIAL AND METHODS: One-stage radical cystectomy and orthotopic ileal W-shaped neobladder creation were performed in 353 male and 97 female patients for invasive bladder cancer. The ureters were reimplanted using a serous-lined extramural tunnel for reflux prevention. Of the patients 344 were evaluable at a mean followup plus or minus standard deviation of 38 +/- 25 months. Evaluation included clinical and radiographic studies to determine functional and oncological outcomes. RESULTS: Four patients (0.8%) died in the hospital. Early complications in 42 patients (9%) were treated conservatively but 3 women underwent vaginal repair of a pouch-vaginal fistula. During the observation period there were 90 oncological failures, of which 3 were isolated urethral recurrence. Late complications included pouch stones in 10 cases, outflow obstruction in 11, mucous retention in 2, adhesive bowel obstruction in 3 and hypercontinence in 9 females. The incidence of daytime and nighttime continence was 93.3% and 80%, respectively. The upper tracts remain unchanged or improved in 96.2% of the reimplanted renal units, while reflux was observed in 3%. CONCLUSIONS: The serous-lined extramural tunnel has proved its efficiency and durability as an antireflux technique.

Optimization of uretero-intestinal anastomosis in urinary diversion: an experimental study in dogs. III. A new antireflux technique for uretero-ileal anastomosis: a serous-lined extramural tunnel.

A new antireflux uretero-ileal reimplantation technique suitable for use with bladder substitutes is presented. This procedure entails creation of a serous-lined extramural tunnel. Following detubularization of the bowel segment, the adjacent flaps are approximated by continuous 4/0 non-absorbable sutures 1.5 cm from the cut edges. The ureters are laid in the trough thus fashioned. "Button-holes" are created in the bowel flaps and a mucosa-to-mucosa uretero-ileal anastomosis is carried out. The mucosal edges of the flaps are then approximated by one layer of continuous 4/0 (PGA) suture resulting in closure of this artificial tunnel. The feasibility and functional outcome of this technique were experimentally investigated in 8 dogs. Follow-up was carried out up to 30 weeks. Assessment by intravenous urography and ascending studies showed that the procedure is an efficient method of providing an unobstructed unidirectional flow of urine.

Further clinical experience with the ileal W-neobladder and a serous-lined extramural tunnel for orthotopic substitution.

OBJECTIVE: To report on the functional results following orthotopic substitution using an ileal W-neobladder with an extramural serous-lined tunnel for reflux prevention. PATIENTS AND METHODS: Sixty men (mean age 46 years) underwent a one-stage radical cystoprostatectomy and an ileal W-neobladder for invasive bladder cancer. The technique entails the creation of two serous-lined extramural tunnels in a detubularized ileal W-bladder fashioned from 40 cm of the terminal ileum. Fifty-one patients were evaluable, with a minimum follow-up of 2 years. Evaluation of patients included clinical, radiographic and urodynamic studies. RESULTS: There was no operative mortality and no gross morbidity. The upper tracts remain unchanged or improved in 97% of the implanted renal units. Reflux was not observed in any patient. The incidence of day and night continence was 90 and 80%, respectively. CONCLUSION: The technique provides a non-obstructed unidirectional uretero-ileal re-implantation, in a low-pressure system constructed from a short ileal segment. No staples were required.

Serous lined extramural ileal valve: a new continent urinary outlet.

PURPOSE: We report the functional results following the use of serous lined extramural valve as an antireflux technique and urinary outlet for continent urinary diversion. MATERIALS AND METHODS: The procedure was performed in 18 men and 5 women. The technique entails fashioning 2 serous lined extramural troughs in a detubularized W-shape ileal reservoir. A tapered ileal segment is embedded in 1 trough as an antireflux valve and the ureters are anastomosed to its proximal end. Another tapered ileal segment or the appendix is embedded in the second trough and acts as a continent cutaneous outlet. RESULTS: No operative or postoperative mortality was observed. One patient had prolonged ileus which was treated conservatively. All patients were evaluable with a mean followup of 19 months. All patients but 1 were continent day and night. No catheterization difficulties were reported. Evacuation intervals were 4 to 5 hours. Radiographic evaluation demonstrated a continent compliant reservoir, stable and straight outlet, and absence of pouch and ureteral reflux. CONCLUSIONS: This procedure is technically feasible, surgically versatile, applicable for urinary diversion or conversion and associated with satisfactory outcome.

Optimization of uretero-intestinal anastomosis in urinary diversion: an experimental study in dogs. II. Influence of exposure to urine on the healing of the ureter and ileum.

The influence of exposure to urine on the ureteric adventitia and the ileal mucous membrane was studied in 10 mongrel dogs. When the ureter was implanted freely into the lumen of the bladder its adventitia became the seat of granulation tissue formation. This is later covered by creeping of transitional epithelium lining the ureter, forming what is in effect a ureteral nipple. Final healing is associated with an unpredictable amount of scarring. Furthermore, it was noted that healing and creeping of the ileal mucous membrane are impeded in the presence of urine. The sum of these effects is that ureters implanted in an open sulcus of the small intestine are not covered by intestinal epithelium, they tend to form spontaneous nipples and their healing is associated with either stenosis or reflux in some 30% of cases.

Optimization of uretero-intestinal anastomosis in urinary diversion: an experimental study in dogs. I. Evaluation of the Le Duc technique.

Experimental evaluation of uretero-ileal reimplantation was carried out in 10 adult mongrel dogs. One half of the ureters (10) were implanted into ileal reservoirs using the classic Le Duc technique. In the other half, the implanted ureters were covered by ileal mucous membrane. Following the classic Le Duc technique, 40% of the reimplanted ureters showed evidence of either reflux and/or stenosis resulting from shortening and fibrosis of the tunnel. The remaining 60% were perfect due to spontaneous nipple formation at the implantation sites rather than to creeping of the intestinal mucosa. In contrast, none of the ureters examined was either refluxing or stenotic. This study outlines the critical importance of covering the implanted ureters with mucosa to avoid the ureteric adventitia being exposed to the irritative effects of urine with subsequent scarring.

Double folded rectosigmoid bladder with a new ureterocolic antireflux technique.

PURPOSE: We present a new method of urinary diversion to the rectum. MATERIALS AND METHODS: Between 1992 and 1995 a new surgical procedure was used in 32 women, 20 men and 12 children 3 to 60 years old (mean age plus or minus standard deviation 35.4 +/- 2.2). The main indication for diversion was invasive bladder carcinoma. Surgery entailed creation of an S-shaped double folded rectosigmoid reservoir with implantation of the ureters via an extramural serous lined antireflux technique. RESULTS: One patient died of a massive pulmonary embolism postoperatively. Followup ranged from 6 to 36 months (mean 19.2 +/- 7.0). During the observation period 6 patients died of local recurrence and/or distant metastasis within 8 months. Of the patients 57 are currently evaluable. All patients are continent during the day with an emptying frequency of 2 to 4 times. Nocturnal enuresis was observed in 4 children who responded favorably to imipramine hydrochloride therapy. Upper urinary tract function was maintained or improved in 95% of the patients. No clinical evidence of acidosis was observed, since all patients were kept on prophylactic oral alkalization. CONCLUSIONS: The procedure can circumvent some of the inherent disadvantages of ureterosigmoidostomy and is a good alternative to orthotopic bladder substitution when the urethra cannot be used.

Primary adenocarcinoma of the urinary bladder: a report of 185 cases.

OBJECTIVE: To evaluate the outcome after the treatment of primary non-urachal vesical adenocarcinoma and to determine the significant prognostic factors. PATIENTS AND METHODS: The records of 185 patients with vesical adenocarcinoma were reviewed. The pathological evaluation included the determination of pathological stage, tumour grade, presence or absence of mucin and its location, evidence of bilharzial infestation and flow-cytometric DNA analysis. The mean follow-up of the treated patients was 3.1 years. Disease-free survival was estimated and the results correlated with patient and tumour characteristics (univariate analysis). Cox's proportional hazards analysis was used to determine prognostic factors. RESULTS: The overall 5-year disease-free survival was 55%; only three factors had a significant impact on survival, the tumour pathological stage and grade, and lymph node involvement. CONCLUSIONS: Radical cystectomy remains the only satisfactory treatment option for primary vesical adenocarcinoma. Tumour stage, grade and lymph node involvement are the only significant prognostic factors.

A case of ileal ureter with proximal antireflux system.

BACKGROUND: The initial experience of constructing a new antirefluxing valve at the uretero-ileal junction with ileal substitution is reported. METHODS: A new antirefluxing valve was constructed at the uretero-ileal junction with ileal substitution by fixing the distal part of the ureter between the psoas muscle and ileal segment (the ileo-psoas tunnel technique). DISCUSSION: The valve created by the technique has been working effectively for preventing the ileo-ureteral reflux. Pre-operative hydronephrosis was improved and the renal function has been well preserved. CONCLUSION: The ileo-psoas tunnel technique is worthwhile when ileal substitution of the ureter is indicated.

The ileopsoas tunnel, a new antireflux technique for ureteroileal reimplantation: an experimental study in dogs.

A new antirefluxing ureteroileal anastomosis is described in which the implanted ureter is placed between the ileal segment and psoas muscle. The procedure was studied in eight experimental dogs, which were observed for 20 weeks. Radiologic and bacteriologic examinations, necropsy assessment and histopathologic results provided evidence that this ileo-psoas tunnel technique allows a unidirectional, non-obstructed flow of urine. The technique could be applied when ileal replacement of the ureter is necessary or in association with continent bladder replacement.

Treatment of urethral defects: skin, buccal or bladder mucosa, tube or patch? An experimental study in dogs.

PURPOSE: We studied 3 graft materials and 2 urethroplasty techniques in 24 adult male mongrel dogs. MATERIALS AND METHODS: The animals were divided into 2 equal groups. In group 1 a 4 cm. segment of perineal urethra was excised and tubed urethroplasty was performed using free full-thickness skin, buccal and bladder mucosa grafts in 4 dogs each. In group 2 a 4 cm. urethral strip was excised and onlay urethroplasty was performed using the same graft materials in 4 dogs each. Retrograde urethrography was done and the animals were sacrificed at week 12. Autopsy specimens were calibrated with a 10Fr catheter. Hematoxylin and eosin stained sections were examined. Masson's trichrome stain was used to determine the extent of fibrosis. RESULTS: Urethral stricture was diagnosed by radiology and confirmed by calibration in 8 of the 12 dogs (66%) in group 1 but in only 1 of the 12 (8%) in group 2 (p <0.004). Buccal mucosa grafts were associated with the lowest stricture rate of 12%, followed by 37% for bladder mucosa and 62% for skin (p <0.2). There was no difference in neovascularization among the 3 grafts. Graft shrinkage was less than 10% for buccal mucosa compared with 20% to 40% for skin and bladder mucosa. The shrinkage rate was similar for the onlay and tube techniques. The intensity of chronic inflammation and fibrosis was highest in the skin grafts. Circumferential fibrosis was noted in association with tubed urethroplasty but not with the onlay technique. CONCLUSIONS: The theoretical advantages of buccal mucosal grafts were pathologically demonstrated. When possible, grafts should be used as an onlay rather than as a complete tube.

Prognostic significance of tumour angiogenesis in schistosoma-associated adenocarcinoma of the urinary bladder.

OBJECTIVE: To report on tumour angiogenesis and its relationship with morphological variables and prognosis in adenocarcinoma of the urinary bladder associated with schistosomiasis. PATIENTS AND METHODS: Fifty-five vesical adenocarcinomas were evaluated from 30 men and 25 women (mean age 47.2 years, sd 8.7, range 30-65) who were followed up after radical cystectomy and urinary diversion for a mean (sd, range) of 61 (43.5, 2.7-159.5) months. Vessels were stained immunohistochemically using an antibody to the platelet endothelial cell-adhesion molecule CD31. Microvessels were counted in active areas of angiogenesis within the tumours (at x250) and the microvessel density (MVD) quantified using the mean of three counts. Treatment failure was defined as death from cancer or the development of local recurrence or distant metastasis. Kaplan-Meier survival curves and Cox's proportional hazard model were used to assess survival. RESULTS: The overall 5- and 10-year survival rates were 57% and 51%, respectively. The presence of lymph node metastasis and high mean vascular density (> 26) were significantly associated with a poor prognosis. The 5-year survival for patients with negative lymph nodes was 66% while no patients with positive nodes survived for 5 years (P < 0.001); the survival was 72% for patients with a low MVD and 33% for those with a high MVD (P = 0.0016). From individual results plotted against vascularity in lymph node-negative patients, there was a significantly better outcome for those with a low MVD (< or = 26; P = 0.0099); this significance was maintained on multivariate analysis. However, there was no significant relationship between angiogenesis and the different clinicopathological factors apart from the grade (P = 0.03); tumour stage, grade and DNA profile had no significant effect on survival in these patients. CONCLUSIONS: These findings suggest that assessing angiogenesis using the MVD provides an independent predictor of survival in patients with adenocarcinoma of the urinary bladder.

Long-term renal morphology and function following enterocystoplasty (refluxing or anti-reflux anastomosis): an experimental study.

OBJECTIVE: To study the morphology and function of the upper urinary tract over the long-term in dogs with an enterocystoplasty and a refluxing or anti-refluxing uretero-intestinal anastomosis. MATERIALS AND METHODS: Subtotal cystectomy and "cup" ileocystoplasty were performed in 13 dogs. The right ureter was implanted into the cystoplasty with a refluxing technique in seven and with an anti-reflux procedure in six dogs. The left renal unit acted as an intact control in 11 dogs, while in two the intramural part of the left ureter was incised to produce reflux. Thus, of the 26 renal units, nine had a refluxing junction (anastomosis), six were anti-refluxing and 11 served as intact controls. Total and separate glomerular filtration rates (GFRs) were measured preoperatively and regularly thereafter, and cystometry, urography and ascending enterocystography were performed. At necropsy, urine was obtained for culture from the cystoplasty and renal pelves, and both kidneys were examined histologically. RESULTS: The cystometric pressure was low in 12 of the 13 dogs: urography showed no obstruction. The fall in separate GFR did not differ significantly among the groups (with and without reflux protection, and control units). Reflux was detected in three of nine renal units with refluxing anastomosis and in three of 11 control units. Bacteriuria was found in the cystoplasty in all dogs; the incidence in the upper urinary tract was seven of eight renal units with a refluxing anastomosis, one in five of those with an anti-refluxing anastomosis and three of nine control units. Pyelonephritis was found in none of the control kidneys, in six of nine kidneys with a refluxing and in two of six with an anti-refluxing anastomosis: it was less severe in the latter. CONCLUSION: Refluxing ureteric implantation in a low-pressure enterocystoplasty was commonly associated with bacteriuria in the upper urinary tract and with pyelonephritis. Thus, anti-reflux implantation was beneficial for renal preservation in this setting.

Long-term outcome analysis of low pressure rectal reservoirs in 33 children with bladder exstrophy.

PURPOSE: The augmented valved rectum and double folded rectosigmoid bladder represent 2 modifications of ureterosigmoidostomy. Both procedures improve continence by lowering the reservoir pressure. We present the outcome of these techniques on the upper urinary tract, continence, metabolic profile, linear growth and bone density. MATERIALS AND METHODS: Between March 1987 and May 1997, 22 boys and 11 girls with bladder exstrophy underwent urinary diversion to a low pressure rectal reservoir. Patient age at surgery ranged from 2 to 13 years (mean 5.4). The augmented valved rectum technique was used in 18 cases and the double folded rectosigmoid bladder method was used in 15. Serial followup ultrasounds were obtained and voiding proctography was performed 1 year postoperatively in all patients. Supine height was measured at last followup in all cases. Serum electrolytes, arterial blood gases, the results of which were plotted on an acid base nomogram, and bone density using dual energy x-ray absorptiometry were measured in all patients. Prophylactic alkalization was administered to all patients. RESULTS: Mean followup is 66 months (range 24 to 148). All patients are continent during the daytime with an emptying frequency of 3 to 5 times, and all are continent at night. No patient experienced pyelonephritis or clinical acidosis. The upper urinary tract was either improved or stabilized in 64 of 66 renal units. No patient had reflux to the upper urinary tract. All patients had normal serum creatinine, sodium, potassium, calcium and phosphorus. Hyperchloremia was noted in 19 of the 33 patients (57%), and arterial blood gases showed subclinical metabolic acidosis in 18 (55%). Supine height measurements at last followup revealed that 19 of the 33 patients (57%) had decreased linear growth (below 3rd percentile). All patients had significant reduction in bone density, and mean for age corrected bone density was 70% (standard deviation 10.9%). Both groups (augmented valved rectum and double folded rectosigmoid bladder) were comparable in regard to age, sex and followup duration. There was no statistically significant difference between the groups in any parameter measured. CONCLUSIONS: The augmented valved rectum and double folded rectosigmoid bladder provide preservation of the upper urinary tract with excellent continence rates. However, prophylactic alkalization and functional isolation of the reservoir do not prevent the long-term metabolic consequences. Subclinical metabolic acidosis and decreased linear growth are to be anticipated in more than 50% of patients. Moreover, significant bone demineralization is to be expected in all of these patients.