Unprecedented conformational flexibility revealed in the ligand-binding domains of the Bovicola ovis ecdysone receptor (EcR) and ultraspiracle (USP) subunits.

The heterodimeric ligand-binding region of the Bovicola ovis ecdysone receptor has been crystallized either in the presence of an ecdysteroid or a synthetic methylene lactam insecticide. Two X-ray crystallographic structures, determined at 2.7 Šresolution, show that the ligand-binding domains of both subunits of this receptor, like those of other nuclear receptors, can display significant conformational flexibility. Thermal melt experiments show that while ponasterone A stabilizes the higher order structure of the heterodimer in solution, the methylene lactam destabilizes it. The conformations of the EcR and USP subunits observed in the structure crystallized in the presence of the methylene lactam have not been seen previously in any ecdysone receptor structure and represent a new level of conformational flexibility for these important receptors. Interestingly, the new USP conformation presents an open, unoccupied ligand-binding pocket.

Abdominal approaches to pelvic prolapse repairs.

PURPOSE OF REVIEW: Abdominal sacrocolpopexy has been considered the gold standard for vaginal vault prolapse repair for several decades. Although transvaginal approaches gained popularity as minimally invasive alternatives, complications related to the use of vaginal mesh have led surgeons to perform these repairs less frequently. By incorporating laparoscopic and robotic techniques into the traditional open abdominal sacrocolpopexy, surgeons can offer the benefits of minimally invasive surgery while avoiding risks of vaginal mesh. This review article aims to evaluate the efficacy and outcomes of abdominal sacrocolpopexy by comparing open, laparoscopic, and robotic assisted laparoscopic surgery. RECENT FINDINGS: The excellent outcomes of open abdominal sacrocolpopexy have repeatedly been shown in published, randomized data. This has been further validated in minimally invasive techniques through randomized data evaluating the outcomes of laparoscopic sacrocolpopexy. Among the various sacrocolpopexy techniques, outcomes are similar among the open, laparoscopic, and robotic approaches. Minimally invasive surgeries have been shown to have advantages in terms of perioperative morbidity. SUMMARY: The superior outcomes of abdominal sacrocolpopexy are available using minimally invasive techniques for pelvic organ prolapse repair. Further research with randomized data is required to establish how these approaches compare to each other. Given the inherent advantages of minimally invasive surgery, robotic or laparoscopic abdominal sacrocolpopexy may become the preferred approach to abdominal pelvic organ prolapse repair.

Color Doppler ultrasound imaging in varicoceles: Is the difference in venous diameter encountered during Valsalva predictive of palpable varicocele grade?

Objective: The clinical grading system for varicoceles is subjective and dependent on clinician experience. Color Doppler ultrasound (US) has not been standardized in the diagnosis of varicoceles. We aimed to determine if US measurement of varicocele could be predictive of World Health Organization (WHO) varicocele grade. Methods: Men who presented for either scrotal pain or infertility to a tertiary men's health clinic underwent physical examination, and varicoceles were graded following WHO criteria (0=subclinical, 1, 2, 3). US was used to measure largest venous diameter in the pampiniform plexus bilaterally at rest and during Valsalva maneuver. Receiver operator characteristic curve analysis was used to determine if resting diameter, diameter during Valsalva, or change in diameter between at rest and during Valsalva provided the highest sensitivity and specificity for determining clinical grade. Threshold values for diameter were determined from these receiver operator characteristic curves. Results: A total of 102 men (50 with clinical varicocele and 52 with subclinical varicocele) were included. Diameter at rest was the best ultrasonographic discriminator between subclinical and clinical varicoceles (area under the curve [AUC]=0.67) with a diameter threshold of 3.0 mm (sensitivity 79%, specificity 42%). Diameter during Valsalva had the greatest AUC for determining clinical Grades 1 versus 2 (AUC=0.57) with diameter threshold of 5.7 mm (sensitivity 71%, specificity 33%). For differentiating between Grades 2 and 3, diameter at rest had the greatest AUC of 0.65 with a threshold of 3.6 mm (sensitivity 71%, specificity 58%). Conclusion: Our results corroborate other studies that have shown a weak correlation between US and clinical grading. The use of diameter during Valsalva was less predictive than diameter at rest and was only clinically significant in differentiating between Grade 1 and 2 varicocele. A standardized method for determining clinically relevant varicoceles on US would allow for improved patient counseling and clinical decision-making.

Implantable Penile Prosthesis for Erectile Dysfunction: Insurance Coverage in the United States.

INTRODUCTION: A manufacturer's benefit verification database was evaluated to ascertain United States health plan insurance coverage for implantable penile prostheses for erectile dysfunction. METHODS: All-payer and employer-sponsored health plan benefit verification databases were queried to determine implantable penile prosthesis approval status. For the all-payer analysis, data by payer were available and presented for 2019-2021 to assess approval status varied by payer and over time. For the employer-sponsored health plan analysis, data by payer were available from 2018-2021. RESULTS: Benefit verification records for the all-payer database were available for 3,167 patients in 2019, 3,016 in 2020, and 2,837 in 2021. Insurance type was preferred provider organization (27.5%), Medicare Advantage (26.9%), Medicare (15.9%), or point-of-service (10.5%). Most patients were approved or verified for implantable penile prosthesis coverage (79.4% in 2019, 79.6% in 2020, and 78.4% in 2021). Coverage was most extensive for government-based insurance (Medicare 98.7%, Medicare Advantage 97.1%, Tricare 100%, and Veterans Affairs 80.0%) but was also favorable for commercial insurance (75.0%). The most common reason for lack of coverage was employer exclusion; the proportion of patients with no coverage due to exclusion increased from 13.5% in 2019 to 17.5% in 2021. Analyses of the employer-sponsored health plan database (n=3,083 patients) showed that 63.1% of patients were approved or verified for coverage and 34.2% did not have coverage due to health plan exclusions. CONCLUSIONS: Approximately 80% of patients had implantable penile prosthesis coverage. Employer exclusion was the most common reason for lagging coverage; rates of employer exclusion increased 29.3% from 2019-2021.

Genetic architecture and evolution of color variation in American black bears.

Color variation is a frequent evolutionary substrate for camouflage in small mammals, but the underlying genetics and evolutionary forces that drive color variation in natural populations of large mammals are mostly unexplained. The American black bear, Ursus americanus (U. americanus), exhibits a range of colors including the cinnamon morph, which has a similar color to the brown bear, U. arctos, and is found at high frequency in the American southwest. Reflectance and chemical melanin measurements showed little distinction between U. arctos and cinnamon U. americanus individuals. We used a genome-wide association for hair color as a quantitative trait in 151 U. americanus individuals and identified a single major locus (p < 10-13). Additional genomic and functional studies identified a missense alteration (R153C) in Tyrosinase-related protein 1 (TYRP1) that likely affects binding of the zinc cofactor, impairs protein localization, and results in decreased pigment production. Population genetic analyses and demographic modeling indicated that the R153C variant arose 9.36 kya in a southwestern population where it likely provided a selective advantage, spreading both northwards and eastwards by gene flow. A different TYRP1 allele, R114C, contributes to the characteristic brown color of U. arctos but is not fixed across the range.

Outcomes of pregnancy following surgery for stress urinary incontinence: a systematic review.

PURPOSE: Although few data have been published on the safety of childbearing after surgery for stress urinary incontinence, a large proportion of physicians recommend that women wait to complete childbearing before pursuing surgical treatment for stress urinary incontinence. We systematically reviewed the available literature to examine the safety of pregnancy after stress urinary incontinence surgery, and to measure the effect of such pregnancy on continence outcomes. MATERIALS AND METHODS: The review was conducted according to the recommendations of the MOOSE (Meta-Analysis of Observational Studies in Epidemiology) group. We performed a systematic review to identify articles published before January 2011 on pregnancy after incontinence surgery. Databases searched include PubMed®, EMBASE® and the Cochrane Review. Our literature search identified 592 titles, of which 20 articles were ultimately included in the review. RESULTS: Data were tabulated from case reports, case series and physician surveys. The final analysis in each category included 32, 19 and 67 patients, respectively. Urinary retention developed during pregnancy in 2 women, 1 of whom was treated with a sling takedown and the other with intermittent catheterization. Of these 2 women 1 also had an episode of pyelonephritis during pregnancy, possibly related to the intermittent catheterization. The incidence of postpartum stress urinary incontinence ranged from 5% to 18% after cesarean delivery and from 20% to 30% after vaginal delivery. CONCLUSIONS: Although the data on outcomes in the literature are limited and further studies need to be performed on the subject, the current data suggests that any increase in risks for pregnancy after surgery for stress incontinence may be small. A low risk of urinary retention during pregnancy may exist. Although some data suggest that cesarean deliveries may result in a lower rate of recurrent stress urinary incontinence than vaginal deliveries, a formal analysis could not be performed with the available data.

The human ileocaecal junction: anatomical evidence of a sphincter.

PURPOSE: The human ileocaecal junction (ICJ) is a major transition zone regulating intestinal transit. Historically, it has often been considered a valve rather than a sphincter. The microscopic anatomy of this junction was studied searching for evidence of an anatomical sphincter and neuromuscular specialisation. METHODS: Ileocaecal specimens were obtained from ten cadavers and five surgical donors (7 male, mean age 81 years, age range 68-94) and examined by histology and immunohistochemistry. Quantitative analyses of muscle thickness and submucosal vascularity were performed together with immunohistochemical studies of innervation and the distribution of interstitial cells of Cajal. RESULTS: The thickness of the muscular layer in both the ileum and the colon increased significantly over a distance of 1 cm leading up to the base of the ileal papilla where it reached a maximum (4.19 ± 2.0 mm) before gradually tapering towards the tip of the papilla. Submucosal vascularity in the ileal papilla was not increased compared to the adjacent ileum or caecum/colon. Neuronal density was less in the caecum and ileal papilla compared to the terminal ileum (P < 0.05). Interstitial cells of Cajal were identified within the myenteric plexus of the ICJ but their density was similar to the adjacent bowel. CONCLUSIONS: A localised muscle thickening at the base of the ileal papilla is consistent with an intrinsic anatomical sphincter. There was no evidence that the ICJ has increased submucosal vascularity or a greater density of innervation compared to the adjacent bowel. The term ileocaecal valve is misleading and should be replaced by ileocaecal junction.

Bipolar Electric Field Pulses in the Martian Magnetosheath and Solar Wind; Their Implication and Impact Accessed by System Scale Size.

The scale size of the plasma boundary region between the sheath and ionosphere in the Martian system is often similar to the gyro-radii of sheath protons, ∼200 km. As a result, ion energization via kinetic structures may play an important role in modifying the ion trajectories and thus be important when evaluating the large-scale dynamics of the Martian system. In this paper, we report observations made with the MAVEN Langmuir Probe and Waves instrument of solitary bipolar electric field structures, and assess their potential role in ion energization in the Martian system. The observed structures appear as short duration (∼0.5 ms) bipolar electric field pulses of ∼1-25 mV/m, and are frequently observed in the upstream solar wind and inside the sheath. The study presented in this paper suggests that the bipolar electric field structures observed at Mars have an average electrostatic potential drop of ∼0.07 V. The estimated upper rate at which these structures could further energize the protons is estimated, assuming the protons gain the full 0.07 eV, to be ∼0.13 eV per gyration, or a change in proton energy of ∼0.3%, and a corresponding change in the gyroradius of ∼0.3 km. These numbers imply that to first order the bipolar structures are not a significant source of ion energization in the Martian magnetosheath.

Synthesis, binding and bioactivity of gamma-methylene gamma-lactam ecdysone receptor ligands: advantages of QSAR models for flexible receptors.

Nuclear hormone receptors, such as the ecdysone receptor, often display a large amount of induced fit to ligands. The size and shape of the binding pocket in the EcR subunit changes markedly on ligand binding, making modelling methods such as docking extremely challenging. It is, however, possible to generate excellent 3D QSAR models for a given type of ligand, suggesting that the receptor adopts a relatively restricted number of binding site configurations or 'attractors'. We describe the synthesis, in vitro binding and selected in vivo toxicity data for gamma-methylene gamma-lactams, a new class of high-affinity ligands for ecdysone receptors from Bovicola ovis (Phthiraptera) and Lucilia cuprina (Diptera). The results of a 3D QSAR study of the binding of methylene lactams to recombinant ecdysone receptor protein suggest that this class of ligands is indeed recognised by a single conformation of the EcR binding pocket.

Efficacy of a preprostatectomy multi-modal penile rehabilitation regimen on recovery of postoperative erectile function.

To evaluate the efficacy of a novel, multi-modal, preoperative approach to postprostatectomy penile rehabilitation (PR), we performed a retrospective review of patients who underwent nerve-sparing robotic-assisted laparoscopic prostatectomy (NS-RALP). All patients were evaluated at a comprehensive, academic sexual medicine clinic between 2016 and 2017. The "prehabilitation" PR group (n = 106) consisted of men who were seen in the pre-op period and began tadalafil and L-citrulline 2 weeks prior to surgery. Vacuum erectile device (VED) therapy was started at 1-month post-op. These interventions were continued throughout the 12-month follow-up period. Individuals refractory to these therapies could start treatment with intracavernosal injections. The postprostatectomy PR group (n = 25) consisted of men who were not seen in the pre-op period and started the above therapies immediately following their first visit. A higher percentage of men in the prehabilitation group reported return of erectile function within 12 months (56% vs. 24%, P = 0.007). The prehabilitation group also showed better compliance with PR (PDE5i [96% vs. 64%, P < 0.001], L-citrulline [93% vs. 49%, P < 0.001], and VED [55% vs. 20%, P < 0.001]). Seventy-eight percent of men who attended 4-5 follow-up visits reported return of erectile function. Our results suggest that men undergoing a preoperative protocol show superior recovery of erectile function following NS-RALP. Further studies with prospective designs are warranted.

Decisional Conflict and Knowledge Among Patients With Varicocele Seeking Treatment for Infertility.

OBJECTIVE: To study disease-specific knowledge and decisional quality in men with varicocele being counseled for infertility. MATERIALS AND METHODS: An instrument designed to measure decisional quality by evaluating disease-specific knowledge, decisional conflict, and the impression that shared decision-making was administered to 92 men identified to have a varicocele seeking their initial infertility consultation. Mean scores on disease-specific knowledge questionnaire, prevalence of decisional conflict, and impact of consultation on preferred infertility treatment were analyzed. RESULTS: Fifty-five percent of patients were found to have decisional conflict. Compared to those with decisional conflict, men without decisional conflict scored higher on the infertility knowledge assessment (63% vs 53% correct) and were more likely to feel that they discussed treatment options with their physician in detail (98% vs 82%) (all P <0.01). Prior to consultation, 28% of all patients preferred assisted reproductive technologies and 2% preferred varicocelectomy as the primary treatment for infertility. Following consultation, 12% and 17% preferred assisted reproductive technologies and varicocelectomy, respectively. The increase in preference for varicocelectomy was greater in men without decisional conflict (5%-31%) than those with conflict (0%-8%) (P = 0.03). CONCLUSION: Infertile men with varicocele have limited knowledge of their disease and high rates of decisional conflict. Before consultation, men with varicoceles showed preference for assisted reproductive technology over varicocele surgery; this trend reversed after consultation. Men with decisional conflict were less likely to prefer varicocelectomy, even after consultation.

Direct-To-Consumer Internet Prescription Platforms Overlook Crucial Pathology Found During Traditional Office Evaluation of Young Men With Erectile Dysfunction.

OBJECTIVE: To determine comorbidities in young men with erectile dysfunction (ED) who are increasingly targeted by direct-to-consumer (DTC) internet platforms that sell phosphodiesterase-5 (PDE-5) inhibitors without comprehensive clinical evaluation; and, further, to characterize the portrayal of DTC platforms by popular news media. METHODS: We retrospectively reviewed all men age ≤40 evaluated for ED at an andrology clinic during January 2016-March 2019 to obtain demographics, exam and lab findings, and treatments. Five news sources were analyzed during the study period to characterize whether articles about DTC platforms were positive, critical, or balanced/neutral. RESULTS: We identified 388 patients, with age 29.5 ± 5.0 years, 15% rate of obesity, 20% prediabetes or diabetes, 54% dyslipidemia, and 20% hypogonadism. Serum lab findings associated with subfertility were found in 11%. Semen analysis was conducted in 64 men, of whom 40% were abnormal. Varicoceles were found in 35%. PDE-5 inhibitor was prescribed to 328 men (88%). Off-label empiric therapies included clomiphene (32.9%) or aromatase inhibitor (12.1%). Testosterone replacement was initiated in 9.7%. Analysis of news coverage revealed 18 articles, of which 61% portrayed DTC platforms exclusively in a positive light. CONCLUSION: Office consultation identified young men with significant comorbidities that would be missed by DTC platforms, which employ only questionnaires for health screening. DTC platforms present themselves as medical authorities without following AUA Guidelines, yet garner mostly positive press coverage. Patients engaging these platforms may falsely believe they are receiving adequate medical assessment. Urologists may do well to incorporate telemedicine to enfranchise young men with evidence-based evaluation.

Decisional Quality Among Patients Making Treatment Decisions for Urolithiasis.

OBJECTIVE: To measure decisional quality, physician loyalty, and treatment preference in patients diagnosed with urolithiasis, a "preference-sensitive" condition, to identify areas of improvement to be addressed by a targeted shared decision-making intervention. METHODS: We identified patients who presented for an initial consultation for urolithiasis from March 2016 to May 2017. Patients completed a 24-item patient experience questionnaire after the consultation which assessed decisional quality domains, physician loyalty, and treatment preference. We summarized treatment preferences before and after the consultation and described the changes. RESULTS: Among the total of 113 patients who met inclusion criteria, 78 (69%) patients chose to participate. Thirty-six (46%) of those patients had evidence of decisional conflict and 42 (54%) did not. Only 1 in 5 patients experiencing decisional conflict reported knowing the benefits and risks of each treatment option. Patients with decisional conflict reported lower perceived shared decision-making, treatment satisfaction, and urolithiasis knowledge. Physician loyalty was lower among patients with decisional conflict as well. Thirty-nine percent of them were 'Promoters' of their urologist, compared to 71% of patients without decisional conflict. Sixty-four percent of patients without decisional conflict identified a treatment preference before consultation, while only 17% of patients who experienced decisional conflict were able to do so. CONCLUSION: Many patients with urolithiasis experience decisional conflict and are unsure of their preferences when making a treatment choice. Uncertainty can correlate with low physician loyalty after consultation. This population may benefit from a shared decision-making intervention that improves decisional quality while incorporating patient-specific preferences.

Time-Driven Activity-Based Costing Analysis of Urological Stone Disease.

INTRODUCTION: The documented increasing incidence of nephrolithiasis in the United States will likely be associated with significant economic impact. Time-driven activity-based costing is an analytical method that has been successfully adapted from industrial analysis for use in health care. Using this costing approach we characterized the cost of 4 stone treatment modalities at our academic medical center, including trial of passage, semirigid ureteroscopy, flexible ureteroscopy and extracorporeal shock wave lithotripsy. METHODS: We developed process mapping for urological evaluation, treatment and followup of renal or ureteral stones less than 10 mm in size for each treatment method. We calculated cost of resources, equipment, disposables, personnel and space used for each step in the process. Cost was based on the capacity of each resource and the amount of time required for the treatment process. RESULTS: The cost for trial of stone passage, $389, was expectedly lower than for surgical interventions and was mainly driven by clinic visit costs. Extracorporeal shock wave lithotripsy and semirigid and flexible ureteroscopy costs were $4,367, $4,830 and $5,356, respectively. Intraoperative disposables and personnel were the top contributors to overall treatment costs. CONCLUSIONS: Conservative management is less costly than surgical interventions. Flexible ureteroscopy is the most expensive of surgical interventions. We describe the first time-driven activity-based cost analysis of stone management to our knowledge. Identifying the main drivers of cost can help to improve the value of urological care and improve future cost-effectiveness analyses.

An anti-TL1A antibody for the treatment of asthma and inflammatory bowel disease.

TL1A is an attractive therapeutic target for the treatment of mucosal inflammation associated with inflammatory bowel disease (IBD) and asthma. Blockade of the TL1A pathway has been shown to reduce inflammatory responses while leaving baseline immunity intact, and to be beneficial in animal models of colitis and asthma. Given the therapeutic potential of blocking this pathway in IBD and asthma, we developed C03V, a human antibody that binds with high affinity to soluble and membrane-bound TL1A. In an assay measuring apoptosis induced by exogenous TL1A, C03V was 43-fold more potent than the next most potent anti-TL1A antibody analyzed. C03V also potently inhibited endogenous TL1A activity in a primary cell-based assay. This potency was linked to the C03V-binding epitope on TL1A, encompassing the residue R32. This residue is critical for the binding of TL1A to its signaling receptor DR3 but not to its decoy receptor DcR3, and explains why C03V inhibited TL1A-DR3 binding to a much greater extent than TL1A-DcR3 binding. This characteristic may be advantageous to preserve some of the homeostatic functions of DcR3, such as TL1A antagonism. In colitis models, C03V significantly ameliorated microscopic, macroscopic and clinical aspects of disease pathology, and in an asthma model it significantly reduced airways inflammation. Notable in both types of disease model was the reduction in fibrosis observed after C03V treatment. C03V has the potential to address unmet medical needs in asthma and IBD.

Investigation of the Christiansen effect in the mid-infrared region for airborne particles.

During measurements of open-path Fourier transform infrared spectra, airborne dust may be present in the infrared beam. We have investigated the feasibility of identifying and quantifying the airborne particulate matter from spectra measured in this way. Although the results showed that analysis of the particulate matter was not able to be performed from these spectra, insight into the size and wavelength dependence of the Christiansen effect at wavelengths where the particles absorb strongly was obtained. Airborne particles larger than or equal to the wavelength of the incident radiation give rise to asymmetrical features in the spectrum caused by the Christiansen effect. However, the transmittance at wavelengths where the refractive index of the particles equals that of the atmosphere never reaches 1.0 because of absorption by the particles. As the particle size becomes much smaller than the wavelength of the incident radiation, the Christiansen effect becomes less pronounced and eventually is not exhibited.

Cost-effectiveness analysis of treatments for metastatic castration resistant prostate cancer.

OBJECTIVE: Treatment options for metastatic castration resistant prostate cancer (mCRPC) have expanded rapidly in recent years. Given the significant economic burden, we sought perform a cost-effectiveness analysis (CEA) of the contemporary treatment paradigm for mCRPC. METHODS: We devised a treatment protocol consisting of sipuleucel-T, enzalutamide, abiraterone, docetaxel, radium-223, and cabazitaxel. We estimated number and length of treatments for each therapy using dosing schedules or progression free survival data from published clinical trials. We estimated treatment cost using billing data and Medicare reimbursement values and performed a CEA. Our analysis assumed US$100,000 per life year saved (LYS) as the threshold societal willingness to pay. RESULTS: Incremental cost-effectiveness ratios (ICER) for strategies incorporating sipuleucel-T that were not eliminated by extended dominance exceeded the societal threshold willingness-to-pay of US$100,000 per LYS, the lowest of which was sipuleucel-T + enzalutamide + abiraterone + docetaxel at US$207,714 per LYS. Enzalutamide + abiraterone + docetaxel exhibited the most favorable ICER among strategies without sipuleucel-T at US$165,460 per LYS. CONCLUSION: Based on the available survival data and current costs of treatment, all treatment strategies greatly exceed a commonly assumed societal willingness-to-pay threshold of US$100,000 per LYS. Improvements in this regard can only come with a reduction in pricing, better tailoring of treatment or significant enhancements in survival with clinical use of treatment combinations or sequences.

Heterogeneity of Outcomes in D'Amico Intermediate-Risk Prostate Cancer Patients after Radical Prostatectomy: Influence of Primary and Secondary Gleason Score.

AIM: The aim of this study was to clarify and examine the outcomes of prostate cancer patients classified as intermediate risk (IR) using the D'Amico risk classification system, specifically focusing on the influence of primary and secondary biopsy Gleason score (BGS). PATIENTS AND METHODS: An institutional review board-approved database of robotic-assisted radical prostatectomies performed after 2006 was stratified by standard D'Amico criteria. IR patients were then sub-stratified by BGS. Pathologic and intermediate-term biochemical disease-free survival (BDFS) outcomes were analyzed. RESULTS: Overall, 1,090 patients were classified as D'Amico low-risk, 896 as IR, and 240 as high-risk. Of the 896 IR patients, 63 had BGS 6, 630 were 3 + 4 = 7, and 203 4 + 3 = 7. Among IR patients, as the BGS increased, there was an increasing likelihood of extracapsular extension (21, 28, and 38%, respectively; p = 0.005), positive surgical margins (14, 26, 31%; p = 0.048), and worse 3-year BDFS (96, 94, 88%; p = 0.01). Multivariable logistic regression and Cox regression analyses confirmed differences among IR groups. CONCLUSION: D'Amico IR patients demonstrate significant heterogeneity in both pathologic outcomes and BDFS. IR patients with a BGS of 6 appear to have similar intermediate-term BDFS as low-risk patients. An increasing BGS from 3 + 3 to 3 + 4 to 4 + 3 results in a higher likelihood of locally-advanced disease and intermediate-term biochemical failure.

Characterisation and cloning of a Na(+)-dependent broad-specificity neutral amino acid transporter from NBL-1 cells: a novel member of the ASC/B(0) transporter family.

Na(+)-dependent neutral amino acid transport into the bovine renal epithelial cell line NBL-1 is catalysed by a broad-specificity transporter originally termed System B(0). This transporter is shown to differ in specificity from the B(0) transporter cloned from JAR cells [J. Biol. Chem. 271 (1996) 18657] in that it interacts much more strongly with phenylalanine. Using probes designed to conserved transmembrane regions of the ASC/B(0) transporter family we have isolated a cDNA encoding the NBL-1 cell System B(0) transporter. When expressed in Xenopus oocytes the clone catalysed Na(+)-dependent alanine uptake which was inhibited by glutamine, leucine and phenylalanine. However, the clone did not catalyse Na(+)-dependent phenylalanine transport, again as in NBL-1 cells. The clone encoded a protein of 539 amino acids; the predicted transmembrane domains were almost identical in sequence to those of the other members of the B(0)/ASC transporter family. Comparison of the sequences of NBL-1 and JAR cell transporters showed some differences near the N-terminus, C-terminus and in the loop between helices 3 and 4. The NBL-1 B(0) transporter is not the same as the renal brush border membrane transporter since it does not transport phenylalanine. Differences in specificity in this protein family arise from relatively small differences in amino acid sequence.