Paclitaxel resistance and the role of miRNAs in prostate cancer cell lines.

PURPOSE: To investigate the expression profiles of 86 miRNAs in paclitaxel-resistant prostate cancer cell lines and to identify the genes that have a role in the development of drug resistance. METHODS: Three prostate cancer cell lines, androgen-dependent VCaP, androgen-independent PC-3 and DU-145, were used to obtain paclitaxel-resistant cells by progressively increasing the concentration of paclitaxel in the culture medium. Viability assays with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium and sulforhodamine B were used to assess the cell resistance level and cytotoxic effects of paclitaxel treatment. Total RNA was isolated from both prostate cancer cell lines and their resistant versions, and cDNA samples were reverse transcribed from total RNA. Selected target genes of miRNAs that showed differences in expression and were estimated to be effective on drug resistance mechanism were analyzed with western blot analysis. RESULTS: Expression study of 86 miRNAs by RT-PCR demonstrated that several of the miRNAs were expressed at different levels in paclitaxel-resistant cells compared to wild-type cells. Moreover, the expression profiles of these miRNAs varied among different prostate cancer cell line types, with 13 miRNAs being up-regulated in the resistant cells. Among these, miR-200b-3p, miR-34b-3p and miR-375 exhibited a marked up-regulation. Further, miR-100-5p showed a prominent increase in paclitaxel-resistant VCaP-R and DU145-R cells. Western blot and RT-PCR studies showed that only the LARP1 and CCND1 genes were over-expressed up to 2-5 times in all paclitaxel-resistant cell lines compared to the other investigated genes. CONCLUSIONS: In this study, the three paclitaxel-resistant prostate cancer cell lines examined showed remarkably different miRNA expression profiles.

Association of B7-H4 gene polymorphisms in urothelial bladder cancer.

BACKGROUND/AIM: We aimed to study polymorphisms of the B7-H4 gene in order to evaluate a possible association in urothelial carcinoma, as it is highly expressed in cancer tissues. MATERIALS AND METHODS: In this study B7-H4 gene rs10754339, rs10801935, and rs3738414 SNPs were studied by PCR-RFLP method in paraffin-embedded tumor specimens from 62 urothelial carcinoma patients and in a control group including 30 patients without bladder cancer. RESULTS: We detected that the rs10754339 polymorphism was more frequent in the cancer patients when compared with the control group (P < 0.05). Only the rs3738414 polymorphism showed a statistically significant difference in frequency between pathologic diagnostic groups. CONCLUSION: The rs10754339 AA genotype distribution was found to have a higher frequency whereas the rs3738414 AG genotype distribution was lower in the bladder cancer group (P < 0.05). None of the genotype distributions showed a significant difference from the control group for the rs10801935 polymorphism. We conclude that B7-H4 has the potential to be a useful prognostic marker in urothelial carcinoma.

Extracorporeal shockwave lithotripsy for ureteral stones: twelve years of experience with 2836 patients at a single center.

PURPOSE: To retrospectively analyze the efficacy of extracorporeal shockwave lithotripsy (SWL) for managing ureteral stones in patients who were treated during a 12-year period at a single center in Turkey. MATERIALS AND METHODS: The study involved 3300 patients who had single ureteral stone and underwent SWL between January 1999 and March 2011. Medical records from 2836 (85%) patients were available for evaluation. Only patients with radiopaque stones of 5- to 15-mm diameter were included. All procedures were carried out by an experienced urologist (ACK). Patients with proximal ureteral calculi were treated in supine position. Those with mid or distal ureteral stones were treated in modified prone position. Persistence of radiologic image of the stone after three SWL sessions or no spontaneous passage of stone fragments after one month of follow-up was defined as treatment failure. Treatment success was defined as radiologically confirmed fragmentation and spontaneous passage of the stone. RESULTS: The success rates for the subgroups with stones located in the proximal, mid, and distal ureter were 85.1%, 83.9%, and 88.4%, respectively (P = .257). The success rates for individuals with smaller stones (≤ 10 mm) in the proximal, mid, and distal ureter were 90%, 85.8%, and 90.4%, respectively (P = .07). The corresponding rates for individuals with larger stones (> 10 mm) were 75.3%, 81.3%, and 81.6%, respectively (P = .09). CONCLUSION: Our retrospective evaluation of this large patient series reveals that SWL is effective for treating stones in the proximal, mid, and distal ureter.

Absorbable versus nonabsorbable graft: outcome of bone anchored male sling for post-radical prostatectomy incontinence.

PURPOSE: We determined the outcome of the bone anchored male sling procedure for stress urinary incontinence in men regarding the graft material used. MATERIALS AND METHODS: A total of 39 men with post-radical prostatectomy incontinence received a perineal bone anchored male sling. Patients with previous salvage external beam radiotherapy and high serum prostate specific antigen, incontinence due to neurogenic or posttraumatic etiology, or previous benign prostatectomy were excluded. Urodynamic evaluation was performed preoperatively. The number of pads daily used by patients was recorded preoperatively and during postoperative visits. To compress the urethra 2 types of materials were used. Absorbable biomaterials were used in the first 12 patients and nonabsorbable material was used in the following 27. RESULTS: Mean patient age +/- SD was 67.3 years (range 50 to 79). The mean duration between radical prostatectomy and male sling surgery was 57.9 +/- 40.4 months (range 5 to 135). The procedure was successful in 26 patients (96.2%) in the nonabsorbable group and in 1 (8.3%) in the absorbable group at a mean followup of 18.9 and 28.8 months, respectively. CONCLUSIONS: The absorbable sling materials that were used for the bone anchored male sling demonstrated disintegration of the material. Autolysis of these absorbable materials removed active compression forces on the urethra after a short period. Nonabsorbable graft is associated with the best outcome following the perineal bone anchored male sling procedure.

Acute effects of sildenafil on Humphrey visual field and intraocular pressure.

PURPOSE: To evaluate the effects of sildenafil on visual field and intraocular pressure in a group of healthy subjects. METHODS: Twenty-eight healthy male volunteers with normal eyes were included in the study. Visual field examinations were performed using FASTPAC 30-2 program (white-on-white and blue-on-yellow) with the Humphrey field analyzer before and one hour after receiving oral 50 and 100 mg sildenafil citrate. RESULTS: The mean age was 51.1 +/- 8.9 years. Mean deviation, pattern standard deviation, short-term fluctuation and corrected pattern standard deviation did not differ significantly among tests both in white-on-white and blue-on-yellow visual field examinations. Changes in intraocular pressure were not statistically significant. CONCLUSIONS: No significant effect of sildenafil was seen on visual field and intraocular pressure in healthy subjects.

Extracorporeal shockwave lithotripsy for solitary distal ureteral stones: does the degree of urinary obstruction affect success?

BACKGROUND AND PURPOSE: Extracorporeal shockwave lithotripsy (SWL) is a safe and effective way to treat stones in the distal ureter, but the impact of urinary obstruction on outcome is not clear. We investigated the relation between the degree of stone-induced urinary obstruction and the outcome of SWL treatment in patients with solitary distal ureteral stones. PATIENTS AND METHODS: A series of 165 consecutive patients with solitary distal ureteral stones underwent SWL (Multimed 9200 Lithotriptor, Elmed Medical Systems, Turkey) between October 2002 and September 2003 at two separate centers. Distal ureteral stones were defined as those located below the lower border of the sacroiliac joint. Patients were divided into four groups according to the degree of stone-induced urinary obstruction: group I (N = 62) had no urinary system dilation; group II (N = 40) had mild dilation, group III (N = 35) moderate dilation, and group IV (N = 28) severe dilation. In addition to degree of obstruction, stone size, average fluoroscopy time, total number of shockwaves applied, number of sessions required to achieve stone-free status, and stone clearance time were recorded. Treatment failure was defined as persistence of fragments after three SWL sessions. RESULTS: Overall, 152 (92.1%) of the patients became stone free after SWL. There were no statistically significant differences among the groups with respect to any of the factors studied. CONCLUSION: In cases where there is a solitary calculus in the distal ureter, the degree of urinary obstruction caused by the stone does not affect the success of stone clearance with SWL.

Effects of carbon dioxide pneumoretroperitoneum on free radical formation in remote organs and use of verapamil as an antioxidant.

BACKGROUND AND PURPOSE: Pneumoretroperitoneum (Prp) acts as an ischemia/reperfusion (I/R) model. Ischemia/reperfusion (I/R) injury causes production of reactive oxygen species, which affect organs remote from the sites of I/R. The aim of this study was to assess the remote organ changes after Prp and to explore the effects of antioxidants. MATERIALS AND METHODS: Eighteen adult rabbits were randomized to three groups, each consisting of six rabbits. Group I (control) underwent balloon dissection of the left retroperitoneal space without gas insufflation. In group II (Prp), carbon dioxide at 10 mm Hg was applied for 2 hours after the balloon dissection (ischemia period) and for 1 hour after desufflation (reperfusion period). In group III (Prp + antioxidant), 5 minutes before the experiment, verapamil at 0.2 mg/kg was given intravenously and the same procedure was employed as in group II. Hepatic, pulmonary, opposite kidney, and treated kidney malondialdehyde (MDA) and reduced glutathione (GSH) levels were evaluated to show response to Prp. RESULTS: Pneumoretroperitoneum exerted oxidative stress on all tissues with an increase of MDA (P < 0.05) and a decrease of GSH (P < 0.05). The verapamil-treated group showed lower values of MDA (P < 0.05) and higher values of GSH (P < 0.05) than group II. CONCLUSION: Pneumoretroperitoneum increased oxidative stress in all remote organs tested. Verapamil reduced the oxidative stress. We concluded that Prp should be employed carefully in patients with limited vital organ capacity. Verapamil administration may be considered for protection against tissue injury attributable to oxidative stress in these patients.

Comparison of changes in tissue oxidative-stress markers in experimental model of open, laparoscopic, and retroperitoneoscopic donor nephrectomy.

PURPOSE: We evaluated the oxidative stress in renal tissue during three types of surgery: open donor nephrectomy (ODN), laparoscopic donor nephrectomy (LDN), and retroperitoneoscopic donor nephrectomy (RDN). The aim was to find out which is the appropriate procedure for harvesting a donor kidney. MATERIALS AND METHODS: Twenty-four New Zealand White rabbits were randomized to four groups, each consisting of six rabbits. Group I (control) was subjected to 180 minutes of anesthesia, and transperitoneal nephrectomy was performed without creation of warm ischemia. In group II (ODN), after 180 minutes of anesthesia, warm ischemia was created for 5 minutes, and nephrectomy was performed. Group III (LDN) was subjected to 5 minutes of warm ischemia after 180 minutes of pneumoperitoneum at 12 mm Hg, and the kidney was removed. In group IV (RDN), after pneumoretroperitoneum at 12 mm Hg for 180 minutes, warm ischemia was created for 5 minutes, and nephrectomy was performed. Renal tissues were analyzed to determine malondialdehyde (MDA) and reduced glutathione (GSH) as oxidative-stress markers. RESULTS: Renal tissue GSH levels were decreased, whereas MDA levels were increased in groups II through IV compared with the control group (p<0.05). There was no statistically significant difference between the ODN, LDN, and RDN groups in the renal oxidative-stress markers. CONCLUSION: No differences were detected in oxidative-stress markers in renal tissue samples between ODN, LDN, and RDN. Therefore, we believe LDN and RDN can be used for live donor kidney harvesting as effectively as ODN without creating greater oxidative stress, which can have deleterious effects on a donor kidney.

Sildenafil citrate does not affect QT intervals and QT dispersion: an important observation for drug safety.

BACKGROUND: Sildenafil is an effective and widely used therapeutic agent for erectile dysfunction. Deaths have been reported due to sildenafil use and most of them are attributed to concurrent use of nitrates. However, the effects of sildenafil on QT intervals, QT dispersion, and the possible risk of ventricular arrhythmia have not been studied before. Our aim in this study was to evaluate the effect of sildenafil citrate on QT intervals and QT dispersion. METHODS: Thirty-six patients with erectile dysfunction were included in this study. Twenty-one patients had coronary artery disease whereas 12 of them also had accompanying diabetes mellitus. Standard 12-lead electrocardiograms (ECG) were recorded three times: before, and at the first and fourth hours of 50 mg sildenafil citrate ingestion. All QT parameters were corrected for heart rate. RESULTS: Mean age of the patients was 54 +/- 12 years. The mean heart rate did not differ significantly between the three ECG examinations. The corrected and uncorrected maximum and minimum QT intervals were not significantly different between the three ECG examinations. The QT dispersion and corrected QT dispersion before and 1 hour and 4 hours after sildenafil ingestion were 31 +/- 9 ms, 36 +/- 10 ms; 32 +/- 11 ms, 37 +/- 14 ms; 27 +/- 8 ms, 32 +/- 9 ms, respectively (P > 0.05). CONCLUSIONS: Sildenafil does not prolong QT intervals or increase QT dispersion in patients with erectile dysfunction. Our results suggest that the risk of ventricular arrhythmia does not increase with ingestion of 50 mg sildenafil.

Effect of retroperitoneoscopic donor nephrectomy on tissue oxidative stress markers in rabbit pneumoretroperitoneum model.

BACKGROUND AND PURPOSE: Laparoscopic donor nephrectomy causes increased renal oxidative stress. There are no data about the effects of the retroperitoneoscopic route. The aim of our study was to evaluate the oxidative stress occurring in renal tissues during retroperitoneoscopic donor nephrectomy in a rabbit model. MATERIALS AND METHODS: Eighteen adult rabbits were randomized to three groups, each consisting of six rabbits. Group I (control) underwent 50-mL balloon dissection of the left retroperitoneal space without CO(2) insufflation. Group II (pneumoretroperitoneum) received a 3-hour CO(2) insufflation at a pressure of 10 mm Hg in the retroperitoneal space after balloon dissection. Group III (pneumoretroperitoneum with warm ischemia), in addition to the procedure applied in Group II, underwent left renal artery clamping for 3 minutes and reperfusion for the next 5 minutes. Bilateral nephrectomy was performed in all animals for analysis of oxidative stress markers. Concentrations of malonyldialdehyde (MDA), protein carbonyl, and reduced glutathione (GSH) were measured in renal tissue samples. RESULTS: The MDA and protein carbonyl content were increased both in the donor (P = 0.004 and P = 0.004, respectively) and in the remaining kidneys (P = 0.009 and P = 0.028, respectively) in Group II compared with Group I. There were no statistically significant increases in oxidative stress markers between Group II and Group III in donor kidneys. However, there were statistically significant decreases in MDA in the remaining kidneys in Group III compared with Group II (P = 0.009). CONCLUSION: Pneumoretroperitoneum causes increased oxidative stress in both donor and remaining kidneys. Short-term warm ischemia and reperfusion do not exert an additive effect on pneumoretroperitoneum-associated oxidative stress in donor kidneys.

Determination of cut-off values for leucocytes and bacteria for urine flow cytometer (UF-100) in urinary tract infections.

Leucocytes and bacteria in urine are the important indices for diagnosing urinary tract infections. Urine specimens from 260 patients were examined for leucocytes and bacteria with the UF-100 flow cytometer (Sysmex Corporation, Japan). The results obtained with the UF-100 were compared with those obtained by bacteriologic cultures of the same urine samples. Bacteriologic samples were found to be positive in 48 urine samples. The best detected cut-off value for bacteria is 3800/microL (senstivity = 73%, specificity = 78%) and 25/microL for leucocytes (sensitivity = 73%, specificity = 92%). In addition, comparison of leucocyte count by manually and with UF-100 shows good correlation (r = 0.91). A flow cytometric urinalysis analyzer operates in a time saving manner, increases throughput and decrease microscopy review rates. It also provides a reliable information to clinicians prior to bacteriologic culture results, in terms of leucocyte and bacteria count.