Urothelial barrier function is maintained by apical membrane plaques and intercellular tight junctions (TJ). Little is known about the composition and regulation of TJ expression in human urothelium. In this study, we have characterised the expression of TJ components in situ and their regulation in an in vitro model of differentiating normal human urothelial (NHU) cells. In normal ureteric urothelium in situ, there was a differentiation-associated profile of claudins 3, 4, 5, 7, ZO1 and occludin proteins. Proliferating NHU cells in vitro expressed predominantly claudin 1 protein and transcripts for claudins 1-5 and 7. Following induction of differentiation by pharmacological activation of PPARγand blockade of EGFR, there was de novo expression of claudin 3 mRNA and protein and downregulation of claudin 2 transcription. There was also a massive increase in expression of claudin 4 and 5 proteins which was due to inhibition of proteasomal degradation of claudin 4 and consequential stabilisation of the claudin 5 heterodimerisation partner. NHU cell differentiation was accompanied by relocalisation of TJ proteins to intercellular junctions. The differentiation-associated development of TJ formation in vitro reflected the stage-related TJ expression seen in situ. This was distinct from changes in TJ composition of NHU cells mediated by increasing the calcium concentration of the medium. Our results imply a role for PPARγ and EGFR signalling pathways in regulating TJ formation in NHU cells and support the hypothesis that TJ development is an integral part of the urothelial differentiation programme.The maintenance of epithelial barrier function requires that the transepithelial passage of water and solutes be tightly regulated. Ion channels and membrane pumps located in the apical and basolateral membrane compartments control transcellular ion transport, whereas tight junctions (TJ), located at the superior aspect of the intercellular junctional complex, control paracellular diffusion (Schneeberger and Lynch, 2004). TJ are composed of cytoplasmic plaque proteins, such as the zonular occludens (ZO) proteins that link the TJ to the cytoskeleton, and integral transmembrane proteins, such as occludin, junctional adhesion molecule (JAM) and claudins that define the properties of the paracellular pore (Tsukita and Furuse, 2002; GonzalezMariscal et al., 2003). The TJ not only limits paracellular movement, but maintain polarity by restricting the movement of proteins and lipids between apical and basolateral membrane compartments. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptThe claudins, which represent a multigene family of some 24 members ranging from 20,000 to 27,000 Mr, are considered to represent the primary seal-forming fibrils of the TJ. Claudins are tetra-spanning proteins, comprising two extracellular loops and short amino and carboxy termini (Schneeberger and Lynch, 2004). The expression of different claudin proteins and the pairing of claudins to form homotypic or heterotypic f...

LKT and IE contributed equally assessed by the acquisition of a transitional cell morphology, a switch from a cytokeratin (CK)13 lo /CK14hi to a CK13 hi /CK14 lo phenotype, expression of claudin 3, 4 and 5 proteins, and induction of uroplakin gene transcription. RESULTSTwo of 12 SUI cell lines showed early senescent changes in culture and were not characterized further; one of seven IC, one of five IDO and a further three SUI cell lines had some evidence of senescence at passage 3. Of the seven IC-derived cell lines, four showed a near normal range of differentiationassociated responses, but the remainder showed little or no response. Most IDO cell lines (four of five) showed a normal differentiation response, but at least three of the 10 SUI cell lines showed some compromise of differentiation potential. CONCLUSIONThis study supports the existence of a subset of patients with IC in whom a failure of urothelial cytodifferentiation might contribute to the disease, and provides a novel platform for investigating the cell biology of urothelium from SUI and other benign dysfunctional conditions.

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Objective: Simulation is now firmly established in modern surgical training and is applicable not only to acquiring surgical skills but also to non-surgical skills and professionalism. A 5-day intensive Urology Simulation Boot Camp was run to teach emergency procedural skills, clinical reasoning, and communication skills using clinical scenario simulations, endoscopic and laparoscopic trainers. This paper reports the educational value of this first urology boot camp. Subjects and methods: Sixteen urology UK trainees completed pre-course questionnaires on their operative experience and confidence level in common urological procedures. The course included seven modules covering basic scrotal procedures, laparoscopic skills, ureteroscopy, transurethral resection of the prostate and bladder tumour, green light laser prostatectomy, familiarisation with common endoscopic equipment, bladder washout to remove clots, bladder botox injection, setting up urodynamics. Emergency urological conditions were managed using scenarios on SimMan®. The main focus of the course was hands-on training using animal models, bench-top models and virtual reality simulators. Post-course assessment and feedback on the course structure and utility of knowledge gained together with a global outcome score was collected. Results: Overall all the sections of feedback received score of over 4.5/5, with the hands-on training on simulators getting the best score 4.8/5. When trainees were asked “The training has equipped me with enhanced knowledge, understanding and skills,” the average score was 4.9/5.0. The vast majority of participants felt they would recommend the boot camp to future junior trainees. Conclusion: This first UK Urology Simulation Boot Camp has demonstrated feasibility and effectiveness in enhancing trainee’s experience. Given these positive feedbacks there is a good reason to expect that future courses will improve the overall skills of a new urology trainee

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Ann R Coll Surg Engl 2007; 89: [799][800][801][802][803] 799 Acute epididymo-orchitis is a common and increasing problem with patients frequently presenting as GP or selfreferrals via the emergency department. Epididymoorchitis is predominantly an acute inflammatory process. However, if left untreated, chronic inflammation may develop with the sequelae of abscess formation, testicular infarction, testicular atrophy and infertility. 1 In men aged >3 5 years, the majority of cases are due to infection with common urinary pathogens and may involve other risk factors such as bladder outlet obstruction and urogenital malformations. In men aged ≤ 35 years who are sexually active, acute epididymo-orchitis may be associated with sexually transmitted organisms and infection of the consort. 2,3 Infection by Chlamydia trachomatis remains the most common bacterial sexually transmitted disease (STD) in the UK and studies have shown that acute epididymoorchitis in young males is frequently associated with C. trachomatis infection. 4,5 The increasing incidence of acute epididymo-orchitis in this younger age group is occurring alongside a nation-wide increase in the transmission rates of C. trachomatis infection and is part of a wider public health crisis of spiralling STD rates in the UK population. 6,7 UROLOGY

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Objectives To evaluate deceased non‐heart beating (DNHB) donors and deceased heart beating (DHB) brain‐stem dead donors, as sources of viable urological tissue for use in biomedical research. To identify sources of viable human bladder tissue as an essential resource for cell biological research aimed at understanding human diseases of the bladder and for developing new tissue engineering and regenerative medicine strategies for bladder reconstruction. Typically, normal human urinary tract tissue is obtained from adult or paediatric surgical patients with benign urological conditions, but few surgical procedures yield useful quantities of healthy bladder tissue for research. Patients and Methods Research ethics committee approval was obtained for collection of donor bladder tissue. Consent for DHB donors was undertaken by the Donor Transplant Coordinators. Tissue Donor Coordinators were responsible for consent for DNHB donors and the retrieval of bladders was coordinated through the National Blood Service Tissue Banking Service. All retrievals were performed by practicing urologists and care was taken to maintain sterility and to minimise bacterial contamination. Two bladders were retrieved from DNHB donors and four were retrieved from DHB donors. Results By histology, DNHB donor bladder tissue exhibited marked urothelial tissue damage and necrosis, with major loss or absence of urothelium. No cell cultures could be established from these specimens, as the urothelial cells were not viable in primary culture. Bladder urothelium from DHB donors was intact, but showed some damage, including loss of superficial cells and variable separation from the basement membrane. All four DHB bladder specimens yielded viable urothelial cells that attached in primary culture, but cell growth was slow to establish and cultures showed a limited capacity to form a functional barrier epithelium and a propensity to senesce early. Conclusions We have shown that normal human bladder urothelial cell cultures can be established and serially propagated from DHB donor bladders. However, our study suggests that rapid post‐mortem changes to the bladder affect the quality and viability of the urothelium, rendering tissue from DNHB donors an inadequate source for urothelial cell culture. Our experience is that whereas patients are willing to donate surgical tissue for research, there is a barrier to obtaining consent from next of kin for retrieved tissues to be used for research purposes.