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The skin is an important barrier against external attacks from bacteria, radicals, or radiations. UV-A radiations cause significant impairment of this barrier, inducing inflammation, oxidative stress, and wrinkle formation, thereby promoting photoaging. Previous studies reported that carnosine, a potent antioxidant, and carbonyl scavenger agent, may prevent photoaging features in the skin of hairless mice exposed to UV-A radiations. In the present study, we used a quantitative proteomic approach to analyze the changes evoked by carnosine in the skin proteome of hairless mice exposed to UV-A. This approach allowed to quantify more than 2480 proteins, among them consistent differences were observed for 89 proteins in UV-A exposed vs control unexposed skins, and 252 proteins in UV-A-exposed skin preventively treated by carnosine (UVAC) vs UV-A. Several functional pathways were altered in the skins of UV-A exposed hairless mice, including the integrin-linked kinase, calcium signaling, fibrogenesis, cell migration and filament formation. An impairment of mitochondrial function and metabolism was observed, with an up-regulation of cytochrome C oxidase 6B1 and NADH: ubiquinone oxidoreductase S8. Skins pre-treated by carnosine were prevented from UV-A induced proteome alterations. In conclusion, our study emphasizes the potency of a proteomic approach to identify the consequences of UV radiations in the skins, and points out the capacity of carnosine to prevent the alterations of skin proteome evoked by UV-A., (Copyright © 2021. Published by Elsevier Inc.)

Objectives: To explore the perspective of urological patients on the possibility to defer elective surgery due to the fear of contracting COVID-19., Methods: All patients scheduled for elective urological procedures for malignant or benign diseases at 2 high-volume centers were administered a questionnaire, through structured telephone interviews, between April 24 and 27, 2020. The questionnaire included 3 questions: (1) In light of the COVID-19 pandemic, would you defer the planned surgical intervention? (2) If yes, when would you be willing to undergo surgery? (3) What do you consider potentially more harmful for your health: the risk of contracting COVID-19 during hospitalization or the potential consequences of delaying surgical treatment?, Results: Overall, 332 patients were included (51.5% and 48.5% in the oncology and benign groups, respectively). Of these, 47.9% patients would have deferred the planned intervention (33.3% vs 63.4%; P < .001), while the proportion of patients who would have preferred to delay surgery for more than 6 months was comparable between the groups (87% vs 80%). These answers were influenced by patient age and American Society of Anesthesiologists score (in the Oncology group) and by the underlying urological condition (in the benign group). Finally, 182 (54.8%) patients considered the risk of COVID-19 potentially more harmful than the risk of delaying surgery (37% vs 73%; P < .001). This answer was driven by patient age and the underlying disease in both groups., Conclusions: Our findings reinforce the importance of shared decision-making before urological surgery, leveraging patients' values and expectations to refine the paradigm of evidence-based medicine during the COVID-19 pandemic and beyond., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objective: The aim of the study is to evaluate how individual and couple's sexuality had changed during the COVID-19 pandemic-related quarantine., Methods: A quantitative correlational research study was conducted, using a web-based survey., Results: About 1576 participants were involved: 1018 women (64.6%) and 558 men (35.4%). A significant decline in the mean well-being scores during the quarantine, compared to before, was reported. A positive correlation between the well-being scores and the number of sexual intercourse (SI) before and during the quarantine was found. The mean number of SI decreased significantly during the quarantine. The main reasons were: poor privacy (43.2%) and lack of psychological stimuli (40.9%). About 1124 respondents (71.3%) did not report sexual desire (SD) reduction. A positive association between SD and SI during the quarantine was found. About 61.2% did not report autoerotism reduction. In those who reported decreased masturbation activity, the main causes were poor privacy (46.4%) and lack of desire (34.7%). We found that men presented lower SD during the quarantine, than women (P < 0.01)., Conclusion: Potentially, the more time available might lead couples to reconnect at an intimate level and to improve their sexuality. However, the majority of quarantined participants experienced reduced number of SI per week, with poor household privacy and lack of psychological stimuli as cited causes, even as a majority did not report reduced autoeroticism., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Carnosine (β-alanyl-L-histidine) is a natural peptide that have been described as a potential pharmacological agent owing to some positive outcomes from several pharmacological tests in animal models of human diseases. However, carnosine has limited activity in humans since the peptide upon absorption is rapidly hydrolyzed in the serum by the enzyme carnosinase (i.e. CN1; E.C. 3.4.13.20). Over the years the main approaches aimed at limiting carnosine hydrolysis have been focused on obtaining CN1-stable derivatives with an increased bioavailability and unmodified or enhanced activity. Only recently the hypothesis of co-administration of carnosine and selective inhibitors of CN1 have been proposed. Such an approach requires reliable methods for screening the effect on carnosine hydrolysis rate operated by CN1 in a throughput scale allowing to test from few compounds up to whole compound libraries. The only assay with such features available in literature relies on ortho-phtalaldehyde (OPA) derivatization of the hydrolysis product (i.e. histidine), followed by a fluorimetric read. Herein, we propose an alternative method based on a direct measurement of the residual substrate by liquid chromatography-mass spectrometry (LC-MS). The assay demonstrated to be reliable since gave results comparable to literature data concerning the hydrolysis rate of carnosine as determined into human serum. Moreover, the method was quite flexible and easily adaptable to a substrate change, as demonstrated by the measurement of the hydrolysis rate of all the natural analogs of carnosine. In this context the data collected for anserine suggest that our method looked more reliable and substrate change can undergo an underestimation of hydrolytic activity in OPA -based assays., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objective: To evaluate semen characteristics and reproductive outcomes after assisted ejaculation methods with fresh in vitro fertilization/intracytoplasmic sperm injection cycles in patients suffering from spinal cord injury (SCI), compared to controls, affected by idiopathic male infertility (non-SCI group)., Methods: SCI patients first underwent penile vibratory stimulation. Men "non-responders" to penile vibratory stimulation underwent electroejaculation. Third, testicular sperm aspiration was proposed., Results: This retrospective monocenter case-control study included 193 couples: 53 couples in SCI group and 140 couples in non-SCI group. Overall, 210 fresh in vitro fertilization/intracytoplasmic sperm injection cycles were performed. Median semen volume of SCI was significantly lower, compared to non-SCI (1.5 mL vs 3.1 mL; P < .01). Median sperm concentration/mL and total sperm count was considerably higher in SCI. Mean sperm progressive motility was significantly lower in SCI (5.0% vs 35.0%; P < .01). Normal fertilization rate was significantly lower in SCI (46.0% vs 71.0%; P < .01). Total fertilization rate was 50.0% and 75%, respectively, in SCI and non-SCI groups. A trend toward higher pregnancy rates per cycle was found in non-SCI (31.4% vs 21.4%), or in the live birth rate, which was 27.1% and 20.0%, respectively. No significant differences were found in pregnancy, miscarriage, and live birth rates per cycle, between the 2 groups., Conclusion: The assisted ejaculation methods in SCI proved to be efficacious and safe to obtain viable sperm for assisted reproductive technologies. Overall, pregnancy and live birth rates were similar to non-SCI patients. Thus, SCI men have the same opportunity to father biological children, compared to men without SCI., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The coronavirus 2019 (COVID-19) pandemic has led to an unprecedented emergency scenario for all aspects of health care, including urology. At the time of writing, Italy was the country with the highest rates of both infection and mortality. A panel of experts recently released recommendations for prioritising urologic surgeries in a low-resource setting. Of note, major cancer surgery represents a compelling challenge. However, the burden of these procedures and the impact of such recommendations on urologic practice are currently unknown. To fill this gap, we assessed the yearly proportion of high-priority major uro-oncologic surgeries at three Italian high-volume academic centres. Of 2387 major cancer surgeries, 32.3% were classified as high priority (12.6% of radical nephroureterectomy, 17.3% of nephrectomy, 33.9% of radical prostatectomy, and 36.2% of radical cystectomy cases). Moreover, 26.4% of high-priority major cancer surgeries were performed in patients at higher perioperative risk (American Society of Anesthesiologists score ≥3), with radical cystectomy contributing the most to this cohort (50%). Our real-life data contextualise ongoing recommendations on prioritisation strategies during the current COVID-19 pandemic, highlighting the need for better patient selection for surgery. We found that approximately two-thirds of elective major uro-oncologic surgeries can be safely postponed or changed to another treatment modality when the availability of health care resources is reduced. PATIENT SUMMARY: We used data from three high-volume Italian academic urology centres to evaluate how many surgeries performed for prostate, bladder, kidney, and upper tract urothelial cancer can be postponed in times of emergency. We found that approximately two-thirds of patients with these cancers do not require high-priority surgery. Conversely, of patients requiring high-priority surgery, approximately one in four is considered at high perioperative risk. These patients may pose challenges in allocation of resources in critical scenarios such as the current COVID-19 pandemic., (Copyright © 2020 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Hyaluronic acid (HA) is physiologically synthesized by several human cells types but it is also a widespread ingredient of commercial products, from pharmaceuticals to cosmetics. Despite its extended use, the precise intra- and extra-cellular effects of HA at low-molecular-weight (LWM-HA) are currently unclear. At this regard, the aim of this study is to in-depth identify and quantify proteome's changes in normal human dermal fibroblasts after 24 h treatment with 0.125, 0.25 and 0.50 % LMW-HA (20-50 kDa) respectively, vs controls. To do this, a label-free quantitative proteomic approach based on high-resolution mass spectrometry was used. Overall, 2328 proteins were identified of which 39 significantly altered by 0.125 %, 149 by 0.25 % and 496 by 0.50 % LMW-HA. Protein networking studies indicated that the biological effects involve the enhancement of intracellular activity at all concentrations, as well as the extracellular matrix reorganization, proteoglycans and collagen biosynthesis. Moreover, the cell's wellness was confirmed, although mild inflammatory and immune responses were induced at the highest concentration. The more complete comprehension of intra- and extra-cellular effects of LMW-HA here provided by an advanced analytical approach and protein networking will be useful to further exploit its features and improve current formulations., Competing Interests: Declaration of Competing Interest This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. The authors state no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aim of this work was to profile, by using an HPLC-MS/MS method, cranberry compounds and metabolites found in human urine after ingestion of a highly standardized cranberry extract (Anthocran®). Two different strategies were adopted for the data analysis: a targeted and an untargeted approach. These strategies allowed the identification of 42 analytes including cranberry components, known metabolites and metabolites hitherto unreported in the literature, including six valerolactones/valeric acid derivatives whose presence in urine after cranberry consumption has never been described before. Absolute concentrations of 26 over 42 metabolites were obtained by using pure available standards. Urine collected at different time points after the last dosage of Anthocran® were tested on the reference strain C. albicans SC5314, a biofilm-forming strain. Fractions collected after 12 h were found to significantly reduce the adhesion and biofilm formation compared to the control (p < 0.05). A similar effect was then obtained by using Anthocran™ Phytosome™, the lecithin formulation containing 1/3 of standardized cranberry extract and formulated to enhance the absorption of the cranberry components. The urinary profile of cranberry components and metabolites in the urine fractions collected at 1 h, 6 h and 12 h after the last capsule intake were then reproduced by using the pure standards at the concentration ranges found in the urine fraction, and tested on C. albicans. Only the mixture mimicking the urinary fraction collected at 12 h and containing as main components, quercetin and 5-(3',4'-dihydroxyphenyl)-γ-valerolactone was found effective thus confirming the ex-vivo results., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

A suitable extraction protocol based on an liquid-liquid extraction with hexane/dimethyl sulfoxide and a GC/MS method were developed and validated to determine the concentration of six prohibited Polycyclic Aromatic Hydrocarbons (PAHs; benzo[a]pyrene; dibenz[a,h]anthracene; benz[a]anthracene; benzo[j]fluoranthene; benzo[k]fluoranthene; chrysene) in lipsticks commissioned by a cosmetic company to a manufacturer. The lipsticks were produced in four different colors. Analyses confirmed the presence of benz[a]anthracene and chrysene only in two colors in a concentration of 9.3-9.4 ng/g. The concentration of PAHs was 250 times lower than what is considered a toxic level on the basis of what reported in the litaraure and guidances for cosmetic ingredients; therefore we could assume that the risk for consumer health was negligeble., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: The hypothesis that renal function could influence oncological outcomes is supported by anecdotal literature., Objective: To determine whether estimated glomerular filtration rate (eGFR) is related to cancer-specific mortality (CSM) in patients who had undergone surgery for renal cell carcinoma (RCC)., Design, Setting, and Participants: A retrospective analysis of 3457 patients who underwent radical (39%) or partial nephrectomy (61%) for cT1-2 RCC between 1990 and 2015., Outcome Measurements and Statistical Analysis: The eGFR was calculated by the Chronic Kidney Disease Epidemiology Collaboration equation. CSM was analyzed in a multivariable competing-risk framework, estimating the subdistribution hazard ratio (SHR) accounting for deaths from other causes. The relationship between eGFR and CSM was investigated from multiple statistical approaches-extended Cox regression with eGFR incorporated as a time-dependent covariate, landmark analysis, and joint modeling. Other predictors were selected by competing-risk random forest method and backward elimination., Results and Limitations: The relationship between eGFR and CSM was graphically described by a linear spline, i.e. a continuous piecewise linear function with two lines joined by a knot. For eGFR treated as a time-dependent covariate, the knot was located at 65ml/min; at landmark analysis with eGFR at the baseline, 12 mo, and last functional follow-up, the knots were 85, 60, and 65ml/min, respectively. In multivariable competing-risk analysis, CSM was associated with eGFR only for values of eGFR below these cutoffs, with SHRs for every 10ml/min of reduction in eGFR of 1.25 (p=0.003), 1.16 (p=0.028), 1.44 (p=0.02), and 1.16 (p=0.042), corresponding to time-dependent eGFR, and eGFR at baseline, 12 mo, and last functional follow-up, respectively. Joint modeling confirmed these results. A retrospective design with inherent biases in data collection represents a limitation., Conclusions: In patients undergoing surgery for RCC, renal function should be preserved in order to improve cancer-related survival., Patient Summary: The relationship between renal function and probability of dying due to renal cancer is complex. The present study found a correlation between glomerular filtration rate and cancer specific mortality that could reconsider the oncological role of renal function in patients undergoing surgery for renal cancer., (Copyright © 2018 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Nowadays, parabens have been replaced by domiphen bromide, which is widely used in pharmaceutical and cosmetic products. The main aim of this study was to investigate stressed degradation products of domiphen bromide by mean of a rapid, specific and reliable LC-ESI MS/MS since phenyl bromination may occur due to the oxidation of bromide counter ion under oxidative conditions. LC-ESI-MS/MS have characterized a new compound, p-bromodomiphen, as the only degradation product and structure elucidation was also confirmed by the synthesis of the standard. Notably, the resulting p-bromodomiphen bromide is more stable then domiphen bromide in oxidizing conditions since no di-bromoderivatives were detected by MS studies; both domiphen and its p-bromo derivative were tested for antibacterial activity and were more effective on Gram positive (Staphylococcus aureus ATCC25923 and Bacillus cereus DSM31) compared to Gram negative bacteria (Escherichia coli ATCC25922 and Pseudomonas aeruginosa DSM22644). In conclusion, stressed degradation studies by LC-ESI-MS/MS have characterized a new compound that comprises an alternative to domiphen bromide since its antimicrobial activity is comparable to, if not better than, the parental compound., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: While partial nephrectomy (PN) represents the standard surgical management for cT1 renal masses, its role for cT2 tumors is controversial. Robot-assisted PN (RAPN) is being increasingly implemented worldwide., Objective: To analyze perioperative, functional, and oncological outcomes of RAPN for cT2 tumors., Design, Setting, and Participants: Retrospective analysis of a large multicenter, multinational dataset of patients with nonmetastatic cT2 masses treated with robotic surgery (ROSULA: RObotic SUrgery for LArge renal mass)., Intervention: Robotic-assisted PN., Outcome Measurements and Statistical Analysis: Patients' demographics, lesion characteristics, perioperative variables, renal functional data, pathology, and oncological data were analyzed. Univariable and multivariable regression analyses assessed the relationships with the risk of intra-/postoperative complications, recurrence, and survival., Results and Limitations: A total of 298 patients were analyzed. Median tumor size was 7.6 (7-8.5) cm. Median RENAL score was 9 (8-10). Median ischemia time was 25 (20-32) min. Median estimated blood loss was 150 (100-300) ml. Sixteen patients had intraoperative complications (5.4%), whereas 66 (22%) had postoperative complications (5% were Clavien grade ≥3). Multivariable analysis revealed that a lower RENAL score (odds ratio [OR] 0.46, 95% confidence interval [CI] 0.21-0.65, p=0.02) and pathological pT2 stage (OR 0.51, 95% CI 0.12-0.86, p=0.001) were protective against postoperative complications. A total of 243 lesions (82%) were malignant. Twenty patients (8%) had positive surgical margins. Ten deaths and 25 recurrences/metastases occurred at a median follow-up of 12 (5-35) mo. At univariable analysis, higher pT stage was predictive of a likelihood of recurrences/metastases (p=0.048). While there was a significant deterioration of renal function at discharge, this remained stable over time at 1-yr follow-up. The main limitation of this study is its retrospective design., Conclusions: RAPN in the setting of select cT2 renal masses can safely be performed with acceptable outcomes. Further studies are warranted to corroborate our findings and to better define the role of robotic nephron sparing for this challenging indication., Patient Summary: This report shows that robotic surgery can be used for safe removal of a large renal tumor in a minimally invasive fashion, maximizing preservation of renal function, and without compromising cancer control., (Published by Elsevier B.V.)

Objective: To examine the different and health-related quality of life (HR-QoL) outcomes between ileal conduit (IC) and ileal orthotopic neobladder (IONB) in patients who underwent radical cystectomy (RC), by using validated self-reported cancer-specific instruments., Patients and Methods: This retrospective, cross-sectional, multicenter cohort study included 148 and 171 patients with either IC or IONB. HR-QoL was evaluated with Quality of Life Core Questionnaire and bladder module (BLM)-30 European Organisation for Research and Treatment of Cancer questionnaires. Baseline HR-QoL scores were dichotomized at the median to give "good" or "poor" score profiles. A matched-pair analysis compared HR-QoL aspects between 79 IC patients and 79 IONB patients., Results: At univariate analysis IONB resulted favorable for physical functioning, emotional functioning, cognitive functioning (CF), fatigue, dyspnea, appetite loss, constipation (CO), and abdominal bloating flatulence (AB). At multivariate analyses, IONB showed better scores for emotional functioning (85 vs 79, P = .023), CF (93 vs 85, P <.001), CO (16 vs 31, P <.001), and AB (12 vs 25, P <.001). A significant worsening of sexual and urinary function was observed for IONB patients in the long-term. At matched-pair analysis, global health status was similar (65 vs 62, P = .385). Significantly better scores were observed in the IONB group for the following items: CF (P = .007), fatigue (P = .003), pain (P = .019), dyspnea (P = .016), CO (P = .001), and AB (P = .00)., Conclusion: IONB and IC after RC were similar in terms of global health status. IONB provides better results in some aspects of HR-QoL related to bowel function, but a worsening of urinary and sexual functions. Further randomized controlled trials are needed to confirm these data., (Copyright © 2017 Elsevier Inc. All rights reserved.)

4-Hydroxynonenal (HNE), an electrophilic end-product deriving from lipid peroxidation, undergoes a heterogeneous set of biotransformations including enzymatic and non-enzymatic reactions. The former mostly involve red-ox reactions on the HNE oxygenated functions (phase I metabolism) and GSH conjugations (phase II) while the latter are due to the HNE capacity to spontaneously condense with nucleophilic sites within endogenous molecules such as proteins, nucleic acids and phospholipids. The overall metabolic fate of HNE has recently attracted great interest not only because it clearly determines the HNE disposal, but especially because the generated metabolites and adducts are not inactive molecules (as initially believed) but show biological activities even more pronounced than those of the parent compound as exemplified by potent pro-inflammatory stimulus induced by GSH conjugates. Similarly, several studies revealed that the non-enzymatic reactions, initially considered as damaging processes randomly involving all endogenous nucleophilic reactants, are in fact quite selective in terms of both reactivity of the nucleophilic sites and stability of the generated adducts. Even though many formed adducts retain the expected toxic consequences, some adducts exhibit well-defined beneficial roles as documented by the protective effects of sublethal concentrations of HNE against toxic concentrations of HNE. Clearly, future investigations are required to gain a more detailed understanding of the metabolic fate of HNE as well as to identify novel targets involved in the biological activity of the HNE metabolites. These studies are and will be permitted by the continuous progress in the analytical methods for the identification and quantitation of novel HNE metabolites as well as for proteomic analyses able to offer a comprehensive picture of the HNE-induced adducted targets. On these grounds, the present review will focus on the major enzymatic and non-enzymatic HNE biotransformations discussing both the molecular mechanisms involved and the biological effects elicited. The review will also describe the most important analytical enhancements that have permitted the here discussed advancements in our understanding of the HNE metabolic fate and which will permit in a near future an even better knowledge of this enigmatic molecule., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The aim of the present investigation was to better understand the pharmacokinetic profile of bilberry (Vaccinium Myrtillus) anthocyanins and the role of glucose transporters (sGLT1 and GLUT2) on their absorption. In particular, the absorption of 15 different anthocyanins contained in a standardized bilberry extract (Mirtoselect ® ) was measured in rats by a validated LC-ESI-MS/MS approach. The plasma concentration peak (Cmax) of 11.1ng/mL was reached after 30min and fasting condition significantly increased the bioavailability of anthocyanins by more than 7 fold in respect to fed rats. Glucose co-administration did not interfere with the overall anthocyanin uptake. Bioavailability of each anthocyanin was then estimated by comparing the relative content in plasma vs extract. The 15 anthocyanins behaved differently in term of bioavailability and both the aglycone and the sugar moiety were found to affect the absorption. For instance, arabinoside moiety was detrimental while cyanidin enhanced bioavailability. Computational studies permitted to rationalize such results, highlighting the role of glucose transporters (sGLT1 and GLUT2) in anthocyanins absorption. In particular a significant correlation was found for the 15 anthocyanins between sGLT1 and GLUT2 recognition and absorption., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Although nephron-sparing surgery (NSS) has demonstrated benefit in terms of renal function preservation, it is unclear whether NSS might also decrease the risk of end-stage renal disease (ESRD) relative to radical nephrectomy (RN). In the current paper, we aimed to report the rate and the predictors of ESRD after surgery, accounting for detailed individual baseline characteristics and comorbidities. A multi-institutional collaboration among five European tertiary care centers allowed study of 2027 patients with normal preoperative renal function and a clinically localized T1abN0M0 renal mass. Cox regression analyses were used to predict the risk of ESRD (defined as the onset of a postoperative estimated glomerular filtration rate <15ml/min per 1.73 m 2 ) after adjusting for the individual baseline risk of developing chronic kidney disease. Univariable ESRD rates at 5 and 10 yr of follow-up were virtually equivalent for patients who underwent NSS (1.5% and 2.5%, respectively) versus RN (1.9% and 2.7%, respectively; hazard ratio [HR]: 0.8; 95% confidence interval [CI], 0.4-1.6). However, diabetes, smoking, uncontrolled hypertension, and other comorbidities were consistently more frequent in the NSS group relative to their RN counterparts. After adjusting for detailed baseline individual characteristics, NSS was shown to have an independent protective effect relative to RN (HR: 0.4; 95% CI, 0.2-0.8; p=0.02) at multivariable analyses., Patient Summary: After accounting for individual baseline characteristics, such as age, diabetes, uncontrolled hypertension, or other comorbidities, partial nephrectomy independently protects against end-stage renal disease and the consequent need for dialysis relative to radical nephrectomy., (Copyright © 2016 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Vinyl sulfones are used for drug design of irreversible inhibitors of cysteine proteases since they are able to alkylate cysteine thiols inside the catalytic pocket of this class of enzymes. Some authors have reported the lack of reactivity towards glutathione as sufficient evidence of the selectivity of such a mechanism. Herein, we demonstrate that some simple molecules containing a vinyl sulfone moiety are not thiol-specific alkylants since they react with some albumin nucleophiles including side chains of Cys34 and His146. Such side-reactions are not desirable for any drug candidate since they limit serum stability, bioavailability and they possibly trigger toxicity mechanisms. In silico predictions, indicate that the compounds tested share similar structural features with reported inhibitors of cysteine proteases, as well as similar poses around the main albumin nucleophiles. Altogether, the data suggest that albumin is better than glutathione for the setup of early in vitro tests probing the selectivity of cysteine protease inhibitors., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Bovine colostrum (BC), the initial milk secreted by the mammary gland immediately after parturition, is widely used for several health applications. We here propose an off-target method based on proteomic analysis to explain at molecular level the potential health benefits of BC. The method is based on the set-up of an exhaustive protein data bank of bovine colostrum, including the minor protein components, followed by a bioinformatic functional analysis. The proteomic approach based on ProteoMiner technology combined to a highly selective affinity chromatography approach for the immunoglobulins depletion, identified 1786 proteins (medium confidence; 634 when setting high confidence), which were then clustered on the basis of their biological function. Protein networks were then created on the basis of the biological functions or health claims as input. A set of 93 proteins involved in the wound healing process was identified. Such an approach also permits the exploration of novel biological functions of BC by searching in the database the presence of proteins characterized by innovative functions. In conclusion an advanced approach based on an in depth proteomic analysis is reported which permits an explanation of the wound healing effect of bovine colostrum at molecular level and allows the search of novel potential beneficial effects., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: To evaluate perioperative results of open (OPN), laparoscopic (LPN), and robot-assisted partial nephrectomies (RAPN) and to identify predictive factors of Trifecta achievement for clinical T1b renal tumors in a multicenter prospective dataset., Methods: Data of 285 patients who had OPN (133), LPN (57), or RAPN (95) for cT1b renal tumors were extracted from the RECORd Project. High-volume centers were defined as ≥50 overall cases of partial nephrectomy per year. Trifecta was defined as simultaneous absence of perioperative complications, negative surgical margins, and ischemia time <25 minutes., Results: The 3 groups had comparable body mass index, preoperative hemoglobin, creatinine and estimated glomerular filtration rate, tumor clinical diameter, and growth pattern. LPN and RAPN were more frequently exclusive of high-volume centers. RAPN showed significantly lower median estimated blood loss compared with OPN and LPN. Trifecta was achieved in 62.4%, 63.2%, and 69.5% of OPN, LPN, and RAPN (P = NS) cases. Median warm ischemia time (WIT) was significantly shorter during OPN than during LPN and RAPN. RAPN had significantly shorter WIT compared with LPN. RAPN was significantly less morbid than OPN regarding intraoperative and postoperative complications. LPN (1.9%) and RAPN (2.5%) showed a lower rate of positive margins compared with OPN (6.8%) (P = NS). At multivariable analysis, exophytic tumor growth pattern, estimated blood loss, and high-volume centers were significant predictive factors for Trifecta achievement., Conclusion: Clinically, T1b renal tumors suitable for NSS can be safely treated by LPN or RAPN in high-volume centers. RAPN allows for significantly lower WIT and estimated blood loss with higher rate of Trifecta achievement compared with LPN., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objective: To externally validate the Young Academic Urologist (YAU) nomogram for the prediction of benign prostatic obstruction (BPO) in patients with lower urinary tract symptoms and benign prostatic enlargement., Materials and Methods: Between January 2013 and September 2014, a consecutive series of patients with lower urinary tract symptoms and benign prostatic enlargement underwent standardized pressure flow studies (PFSs) in 2 tertiary Italian centers. Variables assessed were International Prostatic Symptom Score, Prostate Specific Antigen (PSA), prostate size, transitional zone volume, maximal urinary flow rate (Qmax), postvoid residual urine. BPO was defined as a Schäfer grade ≥ 3 at PFSs. Qmax and transitional zone volume were plotted on the YAU nomogram to predict the presence of BPO. Receiver operating characteristic curve analysis was used to evaluate predictive properties of the nomogram for the final diagnosis of BPO., Results: A total of 449 patients were consecutively enrolled. In those, 310 patients (69%) presented a BPO (Schäfer ≥ 3) at PFSs. The novel YAU nomogram presented an area under the curve of 0.76; 95% confidence interval: 0.72-0.82 for the diagnosis of BPO. At the best cutoff value of 80% (nomogram probability), the sensitivity was 74% and specificity was 79%, the positive predictive value was 89%, and the negative predictive value was 56%., Conclusion: Although further studies are needed to confirm our results, the YAU nomogram was, in our experience, an excellent tool to predict the presence of BPO., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: To evaluate the association between prostate inflammation, metabolic syndrome (MetS), and postoperative lower urinary tract symptoms in patients treated with transurethral resection of the prostate (TURP)., Methods: From April 2011, a consecutive series of patients treated with TURP were prospectively included in this observational study. MetS was defined according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATPIII) criteria. Patients were evaluated at baseline and 1 month postoperative with the International Prostate Symptom Score (IPSS), including the storage IPSS (sIPSS) and voiding IPSS (vIPSS) subscores, the Overactive Bladder questionnaire (OAB-q), and uroflowmetry. Prostate volume was evaluated at baseline. Complications were classified using the modified Clavien system. TURP specimens were examined to define grade, location, and extent of the inflammatory infiltrate according to the standardized classification system of chronic prostatitis and/or chronic pelvic pain syndrome., Results: One hundred and thirty-one subjects were enrolled, in which 54 patients (41.3%) presented with MetS. No differences were observed in terms of preoperative prostate-specific antigen, OAB score, IPSS, vIPSS, sIPSS, Qmax, post void residual, and prostate volume in subjects with and without MetS. An incidence of postoperative complications of 10.6% was recorded: 79% were classified as Clavien type I or II; 21% Clavien IIIb. Of 131 subjects, 97 (74.1%) presented with an inflammatory infiltrate. Patients with MetS presented a high proportion of inflammatory infiltrates compared to patients without MetS (45 of 54; 83% vs 52 of 77; 67%, P = .01). Patients with prostate inflammation presented a 50% risk reduction of postoperative storage urinary symptoms., Conclusion: We confirmed the association between MetS and prostate inflammation.Patients with inflammatory infiltrates mostly benefit from TURP, particularly regarding storage symptoms., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Some reports have suggested that nephron-sparing surgery (NSS) may protect against cardiovascular events (CVe) when compared with radical nephrectomy (RN). However, previous studies did not adjust the results for potential selection bias secondary to baseline cardiovascular risk., Objective: To test the effect of treatment type (NSS vs RN) on the risk of developing CVe after accounting for individual cardiovascular risk., Design, Setting, and Participants: A multi-institutional collaboration including 1331 patients with a clinical T1a-T1b N0 M0 renal mass and normal renal function before surgery (defined as an estimated glomerular filtration rate≥60 ml/min/1.73 m2)., Intervention: RN (n=462, 34.7%) or NSS (n=869, 65.3%) between 1987 and 2013., Outcome Measurement and Statistical Analyses: CVe was defined as onset during the follow-up period of coronary artery disease, cardiomyopathy, hypertension, vasculopathy, heart failure, dysrhythmias, or cerebrovascular disease not known before surgery. Cox regression analyses were performed. To adjust for inherent baseline differences among patients, we performed multivariate analyses adjusting for all available characteristics depicting the overall and cardiovascular-specific profile of the patients., Results and Limitations: When stratifying for treatment type, the proportion of patients who experienced CVe at 1, 5, and 10 yr was 5.5%, 9.9%, and 20.2% for NSS patients compared to 8.7%, 15.6%, and 25.9%, respectively, for RN patients (p=0.001). In multivariate analyses, patients who underwent NSS showed a significantly lower risk of developing CVe compared with their RN counterparts (hazard ratio 0.57, 95% confidence interval 0.34-0.96; p=0.03) after accounting for clinical characteristics and cardiovascular profile. Limitations include the retrospective design of the study because other potential confounders may exist., Conclusions: The risk of CVe after renal surgery is not negligible. Patients treated with NSS have roughly half the risk of developing CVe relative to their RN counterparts. After accounting for clinical characteristics, comorbidities, and cardiovascular risk at diagnosis, NSS independently decreases the risk of CVe relative to RN., Patient Summary: The risk of having a cardiovascular event after renal surgery decreases if a portion of the affected kidney is spared., (Copyright © 2014 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Objective: To evaluate the concordance and prognostic role of histologic variants of bladder urothelial carcinoma in transurethral resection of bladder tumor (TURBT) and radical cystectomy (RC) specimens., Methods: Clinicopathologic information available at the time of RC and follow-up data from 4110 RC specimens, collected between January 2000 and December 2009 at 17 tertiary referral centers were retrospectively analyzed and evaluated for the presence or absence of uncommon variants of bladder urothelial carcinoma. The presence or absence of uncommon variants of bladder urothelial carcinoma was evaluated on previous TURBT specimens of patients undergoing RC. Cox regression was used to assess the impact of these parameters on cancer-specific survival, and the Kaplan-Meier test for disease-free survival was plotted for survival estimate., Results: Of 4110 patients, 579 were found to have uncommon variants of bladder urothelial carcinoma at RC (14.1%), whereas 266 (6.4%) at TURBT. A lack of agreement about uncommon variants was observed between TURBT and RC specimens in the entire population (P <.001). The presence of uncommon variants at TURBT was associated with an increased risk of pathologic upstage (hazard ratio, 3.24; confidence interval, 1.19-6.37; P <.003) and significant decrease in cancer-specific survival and recurrence-free survival (P <.001)., Conclusion: Although the concordance of presence of uncommon histologic variants of urothelial bladder carcinoma between TURBT and RC is low, the presence of uncommon histologic variants of urothelial bladder carcinoma at TURBT is associated with a less favorable clinical outcome., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: To evaluate the association between metabolic syndrome (MetS) and lower urinary tract symptoms (LUTS) in patients with benign prostatic enlargement (BPE)., Materials and Methods: From 2009 onward, a consecutive series of patients with LUTS-BPE were enrolled. Patients were evaluated using the International Prostate Symptom Score (IPSS) and ultrasonographic prostate volume. Body mass index, waist circumference, and blood pressure were measured. Blood samples were collected for prostate-specific antigen levels, fasting glucose levels, triglyceride levels, high-density lipoprotein levels, and testosterone levels. MetS was defined according to Adult Treatment Panel III (ATP III). The risk of detecting LUTS as a function of MetS was evaluated using the logistic regression analysis., Results: A total of 431 patients were enrolled with a median age and prostate-specific antigen level of 67 years (61-73 years) and 3 ng/mL (2.2-4.3 ng/mL), respectively; median body mass index was 27 kg/m2 (25-29 kg/m(2)); median testosterone was 3.9 ng/mL (3.1-4.7 ng/mL); median IPSS was 8 (4-14), median prostate volume was 43 mL (35-56 mL). One hundred three of 431 patients (23.8%) presented with a MetS. Patients with MetS presented a higher IPSS storage subscore (4; interquartile range, 2-7 vs 3; interquartile range 1-7; P = .002). On multivariate analysis, the presence of MetS was associated with an increased risk of an IPSS storage subscore ≥4 (odds ratio, 1.782; 95% confidence interval, 1.045-3.042; P = .030)., Conclusion: In our single-center study, MetS is associated with an increased risk of storage symptoms in patients with BPE. Although these results should be confirmed, and the pathophysiology is yet to be understood, it can be assumed that MetS and its metabolic components should be considered as possible factors involved in LUTS-BPE pathogenesis., (Copyright © 2014 Elsevier Inc. All rights reserved.)

4-Hydroxy-trans-2-nonenal (HNE), an α,β-unsaturated aldehyde generated endogenously by the radical-mediated peroxidation of ω-6 polyunsaturated fatty acids, is a bioactive molecule acting in several physiopathological mechanisms and most of its activity is due to the covalent modification of biomolecules. Although at low and physiological levels HNE acts as an endogenous signaling molecule, a growing bulk of evidence indicates that at high and toxic concentrations, HNE is involved in the onset and propagation of several human diseases. To get more conclusive evidence of HNE as a pathogenetic factor, a pharmacological tool able to inhibit the HNE-induced cellular response is required. Such compound is currently not available, although several molecular strategies have so far been reported with the aim of inhibiting HNE formation or catalyzing its removal. Although most of these are not selective, such strategies have been found to induce several biological responses and would merit further investigation. In this review the various strategies are reported and discussed together with their limits and potentials., (Copyright © 2014 Elsevier Inc. All rights reserved.)

An in vitro high resolution mass spectrometry (MS) method was set-up to test the ability of compounds, mixtures and extracts to inhibit protein carbonylation induced by reactive carbonyl species (RCS). The method consists of incubating the protein target (ubiquitin) with 4-hydroxy-trans-2-nonenal (HNE) in the presence and absence of the tested compound. After 24h of incubation, the reaction is stopped and the protein is analyzed by high-resolution MS. The extent of protein carbonylation is determined by measuring the area of the +11 multicharged peak of the HNE adduct in respect to the native form. The method was validated by measuring the effect of well-known RCS sequestering agents, namely aminoguanidine, pyridoxamine, hydralazine and carnosine, yielding a good reproducibility and the possibility to be automatable. All the compounds were found to dose-dependently inhibit the protein carbonylation with the following order of potency carnosine≈hydralazine≫aminoguanidine>pyridoxamine, as determined by calculating the UC50 values, that is the concentration required to inhibit ubiquitin carbonylation by 50%. A good correlation was found with the results obtained by measuring HNE consumption using an HPLC method optimized by a mobile phase set at pH 7.4, in order to stabilize the eluted adducts. The MS approach was then applied to test the effect of two selected natural extracts on protein carbonylation, i.e. green coffee bean extract and procyanidins from Vitis vinifera. In summary, this paper reports a validated and highly reproducible MS method to test the ability of pure compounds as well as natural extracts to act as protein carbonylation inhibitors., (Copyright © 2013 Elsevier B.V. All rights reserved.)

By using a high resolution top-down and bottom-up approach we identified and characterized the AGEs of beta2-microglobulin (β2-m) formed by incubating the protein in the presence of glucose and of the main reactive carbonyl species. Glucose induced glycation on the N-terminal residue, while glyoxal (GO) and methylglyoxal (MGO) covalently reacted with Arg3. Carboxymethyl (CM-R) and imidazolinone (R-GO) derivatives were identified in the case of GO and carboxyethyl arginine (CE-R) and methyl-imidazolinone (R-MGO) for MGO. Interestingly, α,β-unsaturated aldehydes [4-hydroxy-2-nonenal (HNE); 4-oxo-2-nonenal (ONE); acrolein (ACR)] did not induce any covalent modifications up to 100μM. The different reactivity of β2-m towards the different RCS was then rationalized by molecular modeling studies. The MS method was then applied to fully characterize the AGEs of β2-m isolated from the urine of uremic subjects. CM-R, CE-R and R-MGO were easily identified on Arg3 and their relative abundance in respect to the native protein determined by a semi-quantitative approach. Overall, the AGEs content of urinary β2-m ranged from 0.2 to 1% in uremic subjects. The results here reported offer novel insights and technical achievements for a potential biological role of AGEs-β2-m in pathological conditions., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objective: To compare surgical, pathological, short-term functional data, and complications of endoscopic robotic-assisted simple enucleation (ERASE) and open simple enucleation (OSE)., Methods: We undertook matched-pair analysis (age, tumor size, and preoperative aspects and dimensions used for an anatomical [PADUA] score) of 392 patients treated with simple enucleation (SE) for T1a-T1b renal tumors in our department, including 160 patients in the OSE group and 80 in the ERASE group. Perioperative outcomes were compared with univariate analysis. Variables associated with warm ischemia time (WIT) >25 minutes, complications, and postoperative acute kidney dysfunction (AKD) were assessed with multivariate analysis., Results: The groups were comparable in body mass index (BMI), comorbidity, and preoperative renal function. In the ERASE vs the OSE group, no significant differences resulted regarding WIT (18.5 vs 16.4 minutes, P = .5), complications, transfusion rate, reoperation rate for Clavien grade ≥ 3 complications, and positive surgical margin rate (2.9% vs 2.1%, P = .63). In elective patients, no significant difference resulted in variation of estimated glomerular filtration rate from baseline (8.5 vs 13.9 mL/min, P = .17) and AKD. In the ERASE group, the clamping of renal pedicle was used with a lower frequency (P <.0001), with lower estimated blood loss (EBL), longer operative time, and a 1-day shorter hospitalization (P = .001). On the multivariate analysis, the surgical approach was not independently associated with WIT >25 minutes, postoperative complications, and AKD., Conclusion: The ERASE is a feasible technique with a positive surgical margin rate comparable to OSE; it showed WIT and complication rates similar to the open approach, along with the advantages of mini-invasivity., (Copyright © 2014. Published by Elsevier Inc.)

Background: Approximately 10-20% of recurrences in patients treated with nephrectomy for renal cell carcinoma (RCC) develop beyond 5 yr after surgery (late recurrence)., Objective: To determine features associated with late recurrence., Design, Setting, and Participants: A total of 5009 patients from a multicenter database comprising 13 107 RCC patients treated surgically had a minimum recurrence-free survival of 60 mo (median follow-up [FU]: 105 mo [range: 78-135]); at last FU, 4699 were disease free (median FU: 103 mo [range: 78-134]), and 310 patients (6.2%) experienced disease recurrence (median FU: 120 mo [range: 93-149])., Interventions: Patients underwent radical nephrectomy or nephron-sparing surgery., Outcome Measurements and Statistical Analysis: Multivariable regression analyses identified features associated with late recurrence. Cox regression analyses evaluated the association of features with cancer-specific mortality (CSM)., Results and Limitations: Lymphovascular invasion (LVI) (odds ratio [OR]: 3.07; p<0.001), Fuhrman grade 3-4 (OR: 1.60; p=0.001), and pT stage >pT1 (OR: 2.28; p<0.001) were significantly associated with late recurrence. Based on accordant regression coefficients, these parameters were weighted with point values (LVI: 2 points; Fuhrman grade 3-4: 1 point, pT stage >1: 2 points), and a risk score was developed for the prediction of late recurrences. The calculated values (0 points: late recurrence risk 3.1%; 1-3 points: 8.4%; 4-5 points: 22.1%) resulted in a good-, intermediate- and poor-prognosis group (area under the curve value for the model: 70%; 95% confidence interval, 67-73). Multivariable Cox regression analysis showed LVI (HR: 2.75; p<0.001), pT stage (HR: 1.24; p<0.001), Fuhrman grade (HR: 2.40; p<0.001), age (HR: 1.01; p<0.001), and gender (HR: 0.71; p=0.027) to influence CSM significantly. Limitations are based on the multicenter and retrospective study design., Conclusions: LVI, Fuhrman grade 3/4, and a tumor stage >pT1 are independent predictors of late recurrence after at least 5 yr from surgery in patients with RCC. We developed a risk score that allows for prognostic stratification and individualized aftercare of patients with regard to counseling, follow-up scheduling, and clinical trial design., (Copyright © 2012 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Context: Several randomized controlled trials (RCTs) on phosphodiesterase type 5 inhibitors (PDE5-Is) have showed significant improvements in both lower urinary tract symptoms (LUTS) and erectile dysfunction (ED) in men affected by one or both conditions, without a significant increase in adverse events. However, the results are inconsistent., Objective: Perform a systematic review and meta-analysis of available prospective and cross-sectional studies on the use of PDE5-Is alone or in combination with α1-adrenergic blockers in patients with LUTS/benign prostatic hyperplasia (BPH)., Evidence Acquisition: A systematic search was performed using the Medline, Embase, and Cochrane Library databases through September 2011 including the combination of the following terms: LUTS, BPH, PDE5-Is, sildenafil, tadalafil, vardenafil, udenafil, α-blockers, and α1-adrenergic blocker. The meta-analysis was conducted according to the guidelines for observational studies in epidemiology., Evidence Synthesis: Of 107 retrieved articles, 12 were included in the present meta-analysis: 7 on PDE5-Is versus placebo, with 3214 men, and 5 on the combination of PDE5-Is with α1-adrenergic blockers versus α1-adrenergic blockers alone, with 216 men. Median follow-up of all RCTs was 12 wk. Combining the results of those trials, the use of PDE5-Is alone was associated with a significant improvement of the International Index of Erectile Function (IIEF) score (+5.5; p<0.0001) and International Prostate Symptom Score (IPSS) (-2.8; p<0.0001) but not the maximum flow rate (Q(max)) (-0.00; p=not significant) at the end of the study as compared with placebo. The association of PDE5-Is and α1-adrenergic blockers improved the IIEF score (+3.6; p<0.0001), IPSS score (-1.8; p = 0.05), and Q(max) (+1.5; p<0.0001) at the end of the study as compared with α-blockers alone., Conclusions: The meta-analysis of the available cross-sectional data suggests that PDE5-Is can significantly improve LUTS and erectile function in men with BPH. PDE5-Is seem to be a promising treatment option for patients with LUTS secondary to BPH with or without ED., (Copyright © 2012 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Context: Little is known on the natural history of positive surgical margins (PSMs) in partial nephrectomy (PN). Accumulating data suggest that secondary nephrectomy might not be necessary in all patients with PSMs after PN., Objective: Provide an overview on incidence and risk factors for PSMs after partial nephrectomy and on the rate of local and distant disease recurrence related to PSMs. We also provide recommendations on how to avoid and how to treat PSMs after PN., Evidence Acquisition: A nonsystematic literature research was based on Medline, Scopus, and Web of Science queries on these keywords: nephron-sparing surgery, partial nephrectomy/ies, and margin. Only human studies (original research) published in English were included., Evidence Synthesis: PSMs are present in 0-7% of patients after open PN, in 0.7-4% after laparoscopic PN, and in 3.9-5.7% after robot-assisted PN. The thickness of healthy parenchyma surrounding the tumour is irrelevant as long as complete tumour removal is achieved. The coincidence of a highly malignant tumour and PSM increases the risk of local recurrence. Intermediate follow-up data indicate that the vast majority of patients with PSMs will not experience local or distant tumour recurrence. Frozen-section analysis for evaluation of resection margins during PN is of minor clinical significance, as the surgeon's gross assessment of macroscopically negative margins provides reliable results., Conclusions: PSMs in PN are rare. As indicated by intermediate follow-up data, the majority of patients with PSMs after PN remain without disease recurrence, and a surveillance strategy seems preferable to surgical reintervention., (Copyright © 2011 European Association of Urology. All rights reserved.)

Background: A new edition of the TNM was recently released that includes modifications for the staging system of kidney cancers. Specifically, T2 cancers were subclassified into T2a and T2b (< or =10 cm vs >10 cm), tumors with renal vein involvement or perinephric fat involvement were classified as T3a cancers, and those with adrenal involvement were classified as T4 cancers., Objective: Our aim was to validate the recently released edition of the TNM staging system for primary tumor classification in kidney cancer., Design, Setting, and Participants: Our multicenter retrospective study consisted of 5339 patients treated in 16 academic Italian centers., Intervention: Patients underwent either radical or partial nephrectomy., Measurements: Univariable and multivariable Cox regression models addressed cancer-specific survival (CSS) after surgery., Results and Limitations: In the study, 1897 patients (35.5%) were classified as pT1a, 1453 (27%) as pT1b, 437 (8%) as pT2a, 153 (3%) as pT2b, 1059 (20%) as pT3a, 117 (2%) as pT3b, 26 (0.5%) as pT3c, and 197 (4%) as pT4. At a median follow-up of 42 mo, 786 (15%) had died of disease. In univariable analysis, patients with pT2b and pT3a tumors had similar CSS, as did patients with pT3c and pT4 tumors. Moreover, both pT3a and pT3b stages included patients with heterogeneous outcomes. In multivariable analysis, the novel classification of the primary tumor was a powerful independent predictor of CSS (p for trend <0.0001). However, the substratification of pT1 tumors did not retain an independent predictive role. The major limitations of the study are retrospective design, lack of central pathologic review, and the small number of patients included in some substages., Conclusions: The recently released seventh edition of the primary tumor staging system for kidney tumors is a powerful predictor of CSS. However, some of the substages identified by the classification have overlapping prognoses, and other substages include patients with heterogeneous outcomes. The few modifications included in this edition may have not resolved the most critical issues in the previous version., (Copyright 2010 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

A rapid, sensitive and specific liquid chromatography-electrospray-tandem mass spectrometric (LC-ESI-MS/MS) method for the simultaneous detection and quantitation of methylprednisolone acetate (MPA) and methylprednisolone (MP) in rat plasma, using a triple-stage quadrupole, has been developed and validated. MP-D(2) was used as internal standard (IS) and acetonitrile was added to plasma samples for protein precipitation. After extraction with dichloromethane, the analytes were separated on a C-12 reversed-phase column by isocratic elution (6min at a flow rate 0.2mLmin(-1)) with water containing 0.01% formic acid (A) and acetonitrile (B) (50:50, v/v). Quantitation was performed in positive ion multiple reaction monitoring (MRM) mode by applying the following precursor-to-product ion transitions: MPA m/z 417-->135+161+253; MP m/z 375-->135+161+253; IS m/z 377-->135+161+253. The method, validated over the concentration range 6-600ngmL(-1), has been shown to meet the current requirements of bioananalytical validation, providing satisfactory results in terms of linearity, recovery, intra-day and inter-day precision and accuracy. The lower limit of quantitation (LLOQ) was 6ngmL(-1) for both the analytes (0.080 and 0.072pmol injected for MP and MPA, respectively). The method was successfully applied to monitor the plasma levels of MPA and MP following intra-articular (IA) injections of a low MPA (Depo-Medrol((R))) dose in rats.