A. Peroni

Differential gene expression of cholinergic muscarinic receptor subtypes in male and female normal human urinary bladder

OBJECTIVES To study the mRNA expression of each muscarinic receptor subtype in bladder areas involved in micturition, such as the bladder dome, neck, and trigone. Our study focused on the analysis of the gene expression of muscarinic receptors in the human male and female urinary bladder. Other than the well-known role of bladder parasympathetic innervation, an extensive study of the muscarinic receptor mRNA distribution in male and female urinary bladder is still lacking. METHODS The study was carried out on 5 female (age 56 +/- 10 years) and 5 male (age 70 +/- 9 years) patients. The patients selected for this study did not have any lower urinary tract symptoms, as determined by International Prostate Symptom Score questionnaire. The mRNAs encoding muscarinic receptor subtypes were assessed by reverse transcription-polymerase chain reaction, followed by Southern blot analysis. RESULTS Using a molecular approach, we demonstrated the presence of all muscarinic receptor subtypes in the different urinary bladder areas involved in micturition; in particular, our data indicated that mRNAs encoding muscarinic receptors are largely expressed in all examined bladder areas, both in men and women, although with some remarkable differences and a peculiar distribution. CONCLUSIONS Our results indicate that the pharmacology of the human bladder may be more complex than previously recognized. Furthermore, the choice to study each biopsy as a single sample and not use a pool of tissues allowed us to point out the individual variability between subjects and sex-related differences in the expression profile of muscarinic receptor subtype mRNAs.

Sliding-clip robotic nephropexy

Background Nephroptosis is a clinical condition characterized by an abnormal caudal movement of the kidney that descends more than 2 vertebral bodies (or more than 5 cm) during a position change from supine to upright. In the last decade, the availability of laparoscopic surgery has led to a revival of interest in nephroptosis and nephropexy We describe our technique for transperitoneal laparoscopic robotic-assisted nephropexy. Methods The surgery was performed on a 78-year-old woman with a BMI of 18 and ASA 2. The diagnosis of nephroptosis was preoperatively confirmed by excretory urography in the supine and upright positions. The patient was placed in Trendelemburg position on the left side. The laparoscopic surgery was performed, through a transperitoneal approach, using the daVinci® system: the ptosic kidney was fixed using two separated stiches of non-absorbable prolene 2.0 between the superior surface of the kidney and the quadratus lumborum muscle. The sutures were fixed using Hem-o-lok clips. Results Intra- or post-operative complications have not occurred. The effectiveness of treatment was demonstrated by an intravenous urography 60 days after. Conclusions Robotic-assisted nephropexy is a safe and effective procedure that leads to both clinical and aesthetic excellent results, comparable to the ones obtained with the laparoscopic approach for the treatment of symptomatic renal ptosis. The robotic-assisted nephropexy appears to be easier, with the particular advantages of the intracorporeal suturing and a better intraoperative view; the slidingclips method allows reducing the organ trauma to the minimum.

Short and medium term oncological results after robot-assisted prostatectomy: a comparative prospective non randomized study

Background Our aim was to compare both the perioperative and the short/medium-term post-operative oncological results after radical prostatectomy between two groups of prostate cancer patients: one group treated with radical retropubic prostatectomy (RRP), the other one with robot-assisted laparoscopic prostatectomy (RALP). Methods We collected all the clinical, surgical, pathological and follow-up data of the patients who underwent radical prostatectomy with the two different surgical methods in a dedicated database. The positive surgical margins were defined by the presence of disease at the resection margin marked with China ink. The biochemical persistence of disease was defined as a PSA value >0.1 ng/mL 30 days after surgery, while the biochemical recurrence of the neoplasia was defined as a PSA value >0.2 ng/mL. Results The positive surgical margins (PSM) were identified as follows: 29.3% (RRP) versus 25.4% (RALP) in pT2 cancer (p 0.563), and 63.6% (RRP) versus 50.0% (RALP) in case of >pT2 cancer (p 0.559). If stratified by surgical approach, the results are: 53.3% (RRP) versus 25.6% (RALP) (p <0.001) in case of nerve-sparing technique. The persistence of disease is detectable in 5.4% (RRP) versus 4.6% (RALP), while the data on biochemical recurrence are: 2.7% (RRP) versus 0% (RALP). Conclusions In our experience, in two groups of patients matched for clinic-pathological features who underwent RRP and RALP, oncological results are slightly better in the second group; this figure is even more significant when evaluating surgeries conducted with nerve-sparing technique. In this study we analyzed data of RALP, recently introduced in our center, comparing with RRP, a surgical technique consolidated through the decades. Therefore it can be said that even during the learning curve, RALP provides oncological results comparable to RRP.

MP11-02 OUTCOMES OF A PHASE III RANDOMIZED CONTROLLED TRIAL COMPARING PREVENTIVE VERSUS DELAYED LIGATION OF DORSAL VASCULAR COMPLEX DURING ROBOT-ASSISTED RADICAL PROSTATECTOMY

Primary endpoint was estimated blood loss (EBL); considering significant a difference 330 ml, a sample size of 226 patients were calculated (two-sided α of 0.05 and 80% power). Secondary endpoints were: transfusion rate, positive surgical margins (PSMs), apical PSMs and 1-month PSA and continence (0-1 security pad/day). Differences were compared using Pearson chi-square test or MannWhitney test as appropriate (p<0.05 was considered statistically significant).

MP05-01 MANAGEMENT AND PROGNOSIS OF POSITIVE SURGICAL MARGINS AFTER RADICAL PROSTATECTOMY: RETROSPECTIVE ANALYSIS OF A CONTEMPORARY COHORT

INTRODUCTION AND OBJECTIVES: Positive surgical margins (PSM) after RP are a known factor associated with BCR. Radiation therapy (RT) currently represents an established option for metastasis-free patients. However, the timing of administration is not univocal. The aim of this study is to identify factors related to the indication to adjuvant radiation therapy (aRT) vs salvage (sRT), taking a picture of the contemporary management and prognosis of patients with PSM after radical prostatectomy (RP) at an academic tertiary institution. METHODS: We retrospectively reviewed our perspectivelymaintained database. RP has been performed with an open retropubic approach until 2010, then with a robotic transperitoneal one.All the cases with PSM and adverse pathological features (stage pT3, GS 8) were submitted to a multidisciplinary discussion. The indication to sRT was given if biochemical recurrence (BCR, PSA 0.2 ng/ml), preferably before PSA >0.5 ng/ml. Logistic regression models were used to determine the factors associated with RT indication and BCR in univariate and multivariate analysis. The BCR-free survival was calculated using KaplanMeier method. RESULTS: Out of 789 patients, 197 had PSM (overall prevalence 25,2%), with monofocal involvement in 121 (60.8%) and multifocal in 78 (39.2%). An aRT was indicated in 40 patients (20.3%). Findings are summarized in table 1. Factors independently related to aRT indication were: pathological stage, number of sites of PSM and post-operative PSA. The median follow-up time was 51.1 months (IQR 30.9-69.3). Among the 157 patients for whom aRT was not indicated, 39 experienced a relapse of PSA (prevalence of BCR 24.8%, p not significant). 26 were then treated by sRT, 8 by androgen deprivation therapy, 5 underwent surveillance. Overall, a BCR was found in 46 patients (23.4%) after a median time of 24.0 months (IQR 18.0-36.0). At the last available control 176 patients (89.3%) had a PSA < 0.2 ng/ml (median value 0.02). Only pathological stage was significantly related to the risk of BCR. CONCLUSIONS: In a real-life scenario, the indication to aRT is more restrictive than what recommended by guidelines and is driven by the amount of PSM and a detectable post-operative PSA. No differences in BCR free survival are evident in patients with PSM submitted to aRT vs sRT. Source of Funding: none

MP97-05 OUTCOMES OF PREVENTIVE VS DELAYED LIGATION OF DORSAL VASCULAR COMPLEX DURING RARP: PRELIMINARY RESULTS OF A RANDOMIZED TRIAL

INTRODUCTION AND OBJECTIVES: Little glucose metabolism is generally thought to occur in prostate cancer, leading to low diagnostic accuracy of 18F-fluoro-2-deoxyglucose (FDG) positron emission tomography-computed tomography (PET/CT). Nevertheless, this modality is reportedly useful for identifying high-risk local cancers. We therefore investigated whether local FDG uptake by the prostate reflects the perioperative results of robot-assisted laparoscopic radical prostatectomy (RALP). METHODS: Between November 2012 and August 2016, a total of 248 patients underwent RALP at our institution. Of these, subjects in this study comprised 116 patients in whom FDG-PET/CT was employed for preoperative staging. We retrospectively compared perioperative results between patients, stratified for local FDG uptake in the prostate. Patients who had received preoperative hormone therapy were excluded from the study. FDG uptake was rated based on clinical reports prepared by two radiation diagnosticians. Patient background characteristics, perioperative results and postoperative pathological results were compared between subjects divided into PET-positive and -negative groups. RESULTS: Participants comprised 40 PET-positive subjects and 76 PET-negative subjects. Among the patient background characteristics, mean age was slightly but significantly higher in the PETpositive group (66 years) than in the PET-negative group (64 years; p1⁄40.0485). No significant differences were seen in PSA level, clinical T stage or Gleason Score (GS). Operative time, console time and volume of blood loss also showed no differences between groups, and no patients in either group suffered rectal perforation or required blood transfusion. Postoperative urethral balloon retention time and urinary continence rate at 3 months postoperatively were comparable between groups. Postoperative pathological results showed significantly higher values for the following parameters in the PET-positive group than in the PET-negative group: extraprostatic invasion (45.0% vs 22.4%; p1⁄40.0185); positive margin (30.0% vs 13.2%; p1⁄40.0445); and GS 1⁄48 (52.5% vs 23.7%; p1⁄40.00343). Multivariate analysis also showed that PET positivity tended to be associated with positive margins (odds ratio (OR), 2.45; p1⁄40.0819) and extraprostatic invasion (OR, 2.34; p1⁄40.0529), while GS 1⁄48 was a significant predictor (OR, 3.08; p1⁄40.0208). CONCLUSIONS: In RALP, FDG uptake should be considered a predictor of high-grade disease and a risk factor for positive margins.

Alpha1 Adrenoceptor Subtypes in Human Urinary Bladder: Sex and Regional Comparison

The aim of this work is to report a detailed study of the presence of α1 AR subtypes in human urinary bladder areas involved in the micturition (i.e. detrusor, trigone and neck), investigating whether or not there are differences between sexes. Methods Urinary bladder specimens were obtained as discarded tissue after cystectomy for vesical carcinoma (17 men and 14 women). Molecular characterization of α1 AR subtypes was done by semi-quantitative RT-PCR. The α1 AR protein expression was studied by saturation binding curves using the α1 AR antagonist 125I-HEAT and by western blot. Analysis of data was performed using the GraphPad PRISM 4 software. Results Results obtained indicated that α1 AR proteins were detectable in each bladder area. In both sexes, the detrusor and the neck expressed similar levels of α1 ARs: respectively, detrusor: 14.6 ± 1.2 in men and 13.1 ± 1.1 fmol/mg prot in women; neck: 16.9±3.2 in men and 17.5 ± 4.1 fmol/mg prot in women. In the trigone, significantly higher α1 ARs were found in women compared to men (20.6 ± 1.1 vs 11.7 ± 0.7 fmol/mg prot). Subtype analysis indicated that in women, each area was endowed with mRNA and protein encoding for each α1 AR subtype. The men detrusor expressed α1A and α1D ARs, while in the trigone and the neck, each subtype was present. Conclusions A selective distribution of α1 ARs did exist between sexes and between the different areas involved in micturition, supporting evidence which indicates a role of different α1 AR subtypes in the pathophysiology of the lower urinary tract symptoms.

Evidence for the Presence of α1 Adrenoceptor Subtypes in the Human Ureter

Several studies proposed a role for α1 adrenoceptors (α1 ARs) in the ureteral physiology, indicating that they are present in the ureter; however, few studies have been made to identify α1 AR subtypes present in this area. Thus, this study was carried out to characterize the α1 AR subtype gene and protein expression in proximal, medial and distal region of the human ureter. Methods Molecular characterization of α1 AR subtypes was done by semi-quantitative RT-PCR. The α1 AR protein expression was studied by saturation binding curves using the α1 AR antagonist 125I-HEAT. Analysis of data was performed using the GraphPad PRISM 4 software. Results Analysis of saturation binding curves revealed an heterogeneous distribution of α1 AR binding sites; the Bmax for the distal ureter was indeed 52.5 ±5.4 fmol/mg prot, while a lower similar density of α1 ARs was demonstrated in the medial (25.2 ±1.7 fmol/mg prot) and proximal (23.4 ±0.4 fmol/mg prot) ureters. Molecular characterization of α1 AR subtypes indicated that each receptor was present, although with differences in term of the amount expressed. Conclusions Human ureter was endowed with each α1 AR subtype, although α1D and α1A ARs were prevalent over α1B ARs. Radioligand binding results revealed that there were no significant differences in the Kd between ureteral regions, while an heterogeneous distribution of α1 AR binding sites was detected, with the highest density of α1 ARs in the distal ureter and a lower similar density in the medial and proximal ureter.