By tethering intermediate filaments (IFs) to sites of intercellular adhesion, desmosomes facilitate formation of a supercellular scaffold that imparts mechanical strength to a tissue. However, the role IF–membrane attachments play in strengthening adhesion has not been directly examined. To address this question, we generated Tet-On A431 cells inducibly expressing a desmoplakin (DP) mutant lacking the rod and IF-binding domains (DPNTP). DPNTP localized to the plasma membrane and led to dissociation of IFs from the junctional plaque, without altering total or cell surface distribution of adherens junction or desmosomal proteins. However, a specific decrease in the detergent-insoluble pool of desmoglein suggested a reduced association with the IF cytoskeleton. DPNTP-expressing cell aggregates in suspension or substrate-released cell sheets readily dissociated when subjected to mechanical stress whereas controls remained largely intact. Dissociation occurred without lactate dehydrogenase release, suggesting that loss of tissue integrity was due to reduced adhesion rather than increased cytolysis. JD-1 cells from a patient with a DP COOH-terminal truncation were also more weakly adherent compared with normal keratinocytes. When used in combination with DPNTP, latrunculin A, which disassembles actin filaments and disrupts adherens junctions, led to dissociation up to an order of magnitude greater than either treatment alone. These data provide direct in vitro evidence that IF–membrane attachments regulate adhesive strength and suggest furthermore that actin- and IF-based junctions act synergistically to strengthen adhesion.

(IC) is a chronic bladder inflammatory disease of unknown etiology that is often regarded as a neurogenic cystitis. IC is associated with urothelial lesions, voiding dysfunction, and pain in the pelvic/perineal area, and diet can exacerbate IC symptoms. In this study, we used a murine neurogenic cystitis model to investigate the development of pelvic pain behavior. Neurogenic cystitis was induced by the injection of Bartha's strain of pseudorabies virus (PRV) into the abductor caudalis dorsalis tail base muscle of female C57BL/6J mice. Infectious PRV virions were isolated only from the spinal cord, confirming the centrally mediated nature of this neurogenic cystitis model. Pelvic pain was assessed using von Frey filament stimulation to the pelvic region, and mice infected with PRV developed progressive pelvic pain. Pelvic pain was alleviated by 2% lidocaine instillation into either the bladder or the colon but not following lidocaine instillation into the uterus. The bladders of PRV-infected mice showed markers of inflammation and increased vascular permeability compared with controls. In contrast, colon histology was normal and vascular permeability was unchanged, suggesting that development of pelvic pain was due only to bladder inflammation. Bladder-induced pelvic pain was also exacerbated by colonic administration of a subthreshold dose of capsaicin. These data indicate organ cross talk in pelvic pain and modulation of pain responses by visceral inputs distinct from the inflamed site. Furthermore, these data suggest a mechanism by which dietary modification benefits pelvic pain symptoms.bladder; interstitial cystitis; diet INTERSTITIAL CYSTITIS (IC), also known as painful bladder syndrome, is a chronic bladder inflammatory disease with unknown etiology that afflicts as many as 1 million patients in the United States, with females comprising ϳ90% of patients (21). Symptoms of IC include pelvic and/or perineal pain, urinary frequency, urgency, and nocturia (17,20,32,39). IC is often regarded as a neurogenic cystitis due to voiding dysfunction and the partial efficacy of sacral nerve stimulation or neuropharmacological therapies in some patients that suggests a neuronal component. Supporting this idea, cats are susceptible to feline IC, a disease that closely mimics human IC and is associated with increased activity in the sympathetic nervous system (40). Since IC symptoms can flare in response to certain foods (e.g., tomatoes), dietary modification is commonly employed by IC patients, although evidence of altered urine properties as the mechanism for dietary effects is lacking.One model of IC pathogenesis involves a positive feedback loop, whereby substance P-containing peripheral nerves stimulate mast cells, in turn releasing inflammatory mediators that induce urothelial inflammation (15). Furthermore, histamine release by mast cells feeds back onto peripheral nerves to cause sustained release of substance P and mast cell activation. Consistent with this model, patients with IC show elevated mast cell counts ...

Neuronal growth cone advance was investigated by correlative light and electron microscopy carried out on chick dorsal root ganglion cells. Advance was analyzed in terms of the two principal organelles responsible for protrusive motility in the growth cone – namely, veils and filopodia. Veils alternated between rapid phases of protrusion and retraction. Electron microscopy revealed characteristic structural differences between the phases. Our results provide a significant advance in three respects: first, protruding veils are comprised of a densely branched network of actin filaments that is lamellipodial in appearance and includes the Arp2/3 complex. On the basis of this structural and biomarker evidence, we infer that the dendritic nucleation and/or array-treadmilling mechanism of protrusive motility is conserved in veil protrusion of growth cones as in the motility of fibroblasts; second, retracting veils lack dendritic organization but contain a sparse network of long filaments; and third, growth cone filopodia have the capacity to nucleate dendritic networks along their length, a property consistent with veil formation seen at the light microscopic level but not previously understood in supramolecular terms. These elements of veil and filopodial organization, when taken together, provide a conceptual framework for understanding the structural basis of growth cone advance.

Purpose The primary responsibility of Institutional Review Boards (IRBs) is to protect human research subjects and therefore ensure that studies are conducted in accordance with a standard set of ethical principles. A number of studies have compared the responses from IRBs in multicenter clinical trials involving medical therapies. To date, none have been conducted in trials investigating surgical interventions. The intent of this study was to investigate the consistency of the recommendations issued from one institutional IRB to another in the Minimally Invasive Surgical Therapies (MIST) for benign prostatic hyperplasia (BPH), a multicenter trial with a uniform consent and study protocol. Materials and Methods We obtained the IRB responses from six of the seven participating institutions after the initial submission of the MIST study protocol and classified the responses. We then re-distributed the approved protocols to an IRB at another participating institution and analyzed their review of these protocols. Results We found that both the number and types of responses required for IRB approval of an identical study protocol varied significantly among the participating institutions. We also found that IRB responses were inconsistent in the second review, although all protocols were ultimately approved. Conclusion We conclude that the current system of local IRB review in the context of a multicenter surgical trial is inefficient in the review process and may not provide expertise in overseeing surgical trials. Based on these results, a central surgical IRB may be needed to improve of the ethical review process in multicenter trials.