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The effective storage of urine and its expulsion relies upon the coordinated activity of parasympathetic, sympathetic, and somatic innervations to the lower urinary tract (LUT). At birth, all mammalian neonates lack the ability to voluntary regulate bladder storage or voiding. The ability to control urinary bladder activity is established as connections to the central nervous system (CNS) form through development. The neural regulation of the LUT has been predominantly investigated in adult animal models where comparatively less is known about the neonatal and postnatal neurophysiological development that facilitate urinary continence. Furthermore, congenital neurological or anatomical defects can adversely affect both storage and voiding functions through postnatal development and into adulthood, leading to secondary conditions including vesicoureteral reflux, chronic urinary tract infections, and end-stage renal disease. Therefore, the aim of the review is to provide the current knowledge available on neurophysiological regulation of the LUT through pre- to postnatal development of human and animal models and the consequences of congenital anomalies that can affect LUT neural function.

Benign prostatic hyperplasia (BPH) is a feature of ageing males. Up to half demonstrate bladder outlet obstruction (BOO) with associated lower urinary tract symptoms (LUTS) including bladder overactivity. Current therapies to reduce obstruction, such as α1-adrenoceptor antagonists and 5α-reductase inhibitors, are not effective in all patients. The phosphodiesterase-5 inhibitor (PDE5I), tadalafil, is also approved to treat BPH and LUTS suggesting a role for nitric oxide (NO•), soluble guanylate cyclase (sGC), and cGMP signalling pathways. However, PDE5I refractoriness can develop for reasons including nitrergic nerve damage and decreased NO• production, or inflammation-related oxidation of the sGC haem group, normally maintained in a reduced state by the cofactor, cytochrome-b5-reductase 3 (CYB5R3). sGC activators, such as cinaciguat (BAY 58-2667), have been developed to enhance sGC activity in the absence of NO• or when sGC is oxidised. Accordingly, their effects on the prostate and LUT function of aged mice were evaluated. Aged mice (≥24 months) demonstrated a functional BPH/BOO phenotype, compared to adult animals (2-12 months), with low, delayed voiding responses and elevated intravesical pressures as measured by telemetric cystometry. This was consistent with outflow tract histological and molecular data that showed urethral constriction, increased prostate weight, greater collagen deposition and cellular hyperplasia. All changes in aged animals were attenuated by daily oral treatment with cinaciguat for two weeks, without effect on serum testosterone levels. Cinaciguat had only transient (1 h) cardiovascular effects with oral gavage suggesting a positive safety profile. The benefit of cinaciguat was suggested by its reversal of an overactive cystometric profile in CYB5R3 smooth muscle knock-out mice that mirrors a profile of oxidative dysfunction where PDE5I may not be effective. Thus, the aged male mouse is a suitable model for BPH-induced BOO and cinaciguat has a demonstrated ability to reduce prostate-induced obstruction and consequent effects on bladder function. This article is protected by copyright. All rights reserved.

OBJECTIVE: To assess the functional role of Hyperpolarization-activated cyclic nucleotide-gated gated channel (HCN) subtypes in the aging bladder phenotype characterized by diminished bladder volume sensation (BVS) with or without the detrusor instability (DI). METHODS: Expression of HCN subtypes was examined by quantitative RT-PCR and Western blot in aged male Fisher 344 rats (n=15) and young rats (n=15). Nocturnal urination and awake cystometry (CMG) were assessed in presence and absence of a steady state HCN channel blockade achieved with daily oral gavage of vehicle or Ivabradine (HCN blocker) 6mg/kg for 7 days. RESULTS: The association of BVS with the age-related downregulation (~30%) of cAMP sensitive HCN1, HCN2 subtypes, and (~50%) upregulation of cAMP insensitive HCN3 subtype is evinced by a doubling in the mean urine volume of nocturnal voids (0.82 ±0.22 mL vs 0.41 ±0.12 mL; n=10; p < 0.05) predicting an age-related rise in micturition volume threshold (p

To assess whether quantitative T1 relaxometry can measure permeability, chronic inflammation and mural thickening of mouse bladder wall. Adult female C57BL6 mice unexposed to radiation (controls) or 40 wk postirradiation of 10 Gy were scanned at 9.4 T before and after instillation (0.1 mL) of aqueous, novel contrast mixture (NCM) containing 4 mM gadobutrol and 5 mM ferumoxytol. Rapid acquisition with refocused echo (RARE) sequence was used with variable repetition times (TR). Pixel-wise maps of T1 relaxation times for the segmented bladder wall layers were generated from voxel-wise, nonlinear least square data fitting of TR-dependent signal intensity acquired with TR array of 0.4–10 s followed by the histology of harvested bladder. Significant differences between precontrast and postcontrast T1 (ΔT1) were noted in urothelium and lamina propria of both groups but only in detrusor of irradiated group ( P < 0.001; 2-way ANOVA). Nearly twofold higher gadobutrol permeability (550 ± 73 vs. 294 ± 160 μM; P < 0.01) derived as per 1/ΔT1 = r1. [C] in urothelium of irradiated group. Inflammation and bladder wall thickening (0.75 ± 0. vs. 0.44 ± 0.08 mm; P < 0.001) predicted by MRI was subsequently confirmed by histology and altered expression of CD45 and zonula occludens-1 (ZO-1) relative to controls. NCM enhanced MRI relies on the retention of large molecular weight ferumoxytol in lumen for negative contrast, while permeation of the non-ionic, small molecular weight gadobutrol through ZO-1 generates positive contrast in bladder wall for virtual measurement of paracellular permeability and assessment of chronic inflammation in thin and distensible bladder wall, which is also defined by its variable shape and location within pelvis.

INTRODUCTION AND OBJECTIVE:Radiation-induced cystitis (RC) is a progressive debilitating disorder and its most prevalent phase is the chronic phase that occurs months and years after radiation expo...

PURPOSE: Transient receptor potential cation channel subfamily M member 4 (TRPM4) has been shown to play a key role in detrusor contractility under physiological conditions. In this study we investigated the potential role of TRPM4 in detrusor overactivity following spinal cord transection (SCT) in mice. METHODS: TRPM4 expression and function were evaluated in bladder tissue with or without the mucosa from spinal intact (SI) and SCT female mice (T8-T9 vertebra; 1–28 days post SCT) using PCR, western blots, immunohistochemistry and muscle strip contractility techniques. RESULTS: TRPM4 was expressed in the urothelium (UT) and detrusor smooth muscle (DSM) and was upregulated after SCT. Expression levels peaked 3–7 days post SCT in both the UT and DSM. Pharmacological block of TRPM4 with the antagonist, 9-Phenanthrol (30 µM) greatly reduced spontaneous phasic activity that developed after SCT, regardless of the presence or absence of the mucosa. CONCLUSIONS: Detrusor overactivity following spinal cord injury leads to incontinence and/or renal impairment, and represents a major health problem for which current treatments are not satisfactory. Augmented TRPM4 expression in the bladder after chronic SCT supports the hypothesis that TRPM4 channels play a role in DSM overactivity following SCT. Inhibition of TRPM4 may be beneficial for improving detrusor overactivity in SCI.

New findings What is the central question of this study? Overactive bladder is associated with enhanced spontaneous contractions, but their origins are unclear. The aim of this study was to characterize the accompanying ATP transients. What is the main finding and its importance? Spontaneous detrusor contractions were accompanied by transient increases of ATP, and their appearance was delayed by previous activation of efferent nerves to the detrusor. This indicates that spontaneous ATP release from nerve terminals supports spontaneous contractions. ATP is a functional excitatory neurotransmitter in human bladder only in pathologies such as overactive bladder. A potential drug target is revealed to manage this condition. Abstract Spontaneous contractions are characteristic of the bladder wall, but their origins remain unclear. Activity is reduced if the mucosa is removed but does not disappear, suggesting that a fraction arises from the detrusor. We tested the hypothesis that spontaneous detrusor contractions arise from spontaneous ATP release. Guinea-pig detrusor strips, without mucosa, were superfused with Tyrode solution at 36°C. Preparations were subjected to electrical field stimulation (EFS; 3 s trains at 90 s intervals) to produce nerve-mediated contractions, abolished by 1 µm TTX. Amperometric ATP electrodes on the preparation surface recorded any ATP released. Spontaneous contractions and ATP transients were recorded between EFS trains. Nerve-mediated contractions were attenuated by atropine and α,β-methylene ATP; in combination, they nearly abolished contractions, as did nifedipine. Contractions were accompanied by ATP transients that were unaffected by atropine but inhibited by TTX and greatly attenuated by nifedipine. Spontaneous contractions were accompanied by ATP transients, with a close correlation between the magnitudes of both transients. ATP and contractile transients persisted with TTX, atropine and nifedipine. Immediately after a nerve-mediated contraction and ATP transient, there was a longer interval than normal before spontaneous activity resumed. Spontaneous contractions and ATP transients are proposed to arise from ATP leakage from nerve terminals innervating the detrusor. Extracellular ATP has a greater functional significance in humans who suffer from detrusor overactivity (spontaneous bladder contractions associated with incontinence) owing to its reduced hydrolysis at the nerve-muscle interface. This study shows the origin of spontaneous activity that might be exploited to develop a therapeutic management of this condition.

Loss of bladder control is a challenging outcome facing patients with spinal cord injury (SCI). We report that systemic blocking of pro–nerve growth factor (proNGF) signaling through p75 with a CNS-penetrating small-molecule p75 inhibitor resulted in significant improvement in bladder function after SCI in rodents. The usual hyperreflexia was attenuated with normal bladder pressure, and automatic micturition was acquired weeks earlier than in the controls. The improvement was associated with increased excitatory input to the spinal cord, in particular onto the tyrosine hydroxylase–positive fibers in the dorsal commissure. The drug also had an effect on the bladder itself, as the urothelial hyperplasia and detrusor hypertrophy that accompany SCI were largely prevented. Urothelial cell loss that precedes hyperplasia was dependent on p75 in response to urinary proNGF that is detected after SCI in rodents and humans. Surprisingly, death of urothelial cells and the ensuing hyperplastic response were beneficial to functional recovery. Deleting p75 from the urothelium prevented urothelial death, but resulted in reduction in overall voiding efficiency after SCI. These results unveil a dual role of proNGF/p75 signaling in bladder function under pathological conditions with a CNS effect overriding the peripheral one.

BACKGROUND AND PURPOSETo characterise the molecular mechanisms that determine variability of atropine-resistance of nerve-mediated contractions in human and guinea-pig detrusor smooth muscle EXPERIMENTAL APPROACHAtropine-resistance of nerve-mediated contractions, and the role of P2X1 receptors, was measured in isolated preparations from guinea-pigs and also humans with or without overactive bladder syndrome, from which the mucosa was removed. Nerve-mediated ATP release was measured directly with amperometric ATP-sensitive electrodes. Ecto-ATPase activity of guinea-pig and human detrusor samples was measured in vitro by measuring the concentration-dependent rate of ATP breakdown. The transcription of ecto-ATPase subtypes in human samples was measured by qPCR.KEY RESULTSAtropine resistance was greatest in guinea-pig detrusor, absent in human tissue from normally-functioning bladders and intermediate in human overactive bladder. Greater atropine resistance correlated with reduction of contractions by the ATP-diphospho-hydrolase apyrase, directly implicating ATP in their generation. E-NTPDase-1 was the most abundantly transcribed ecto-ATPase of those tested and transcription was reduced in tissue from human overactive, compared to normal, bladders. E-NTPDase-1 enzymatic activity was inversely related to the magnitude of atropine resistance. Nerve-mediated ATP release was continually measured and varied with stimulation frequency over the range 1-16 Hz.CONCLUSION AND IMPLICATIONSAtropine-resistance in nerve-mediated detrusor contractions is due to ATP release and its magnitude is inversely related to E-NTPDase-1 activity. ATP is released under different stimulation conditions compared to acetylcholine that implies different routes for their release.

OBJECTIVE: To evaluate whether approved gastroprokinetic agent, acotiamide exerts a direct excitatory effect in bladder to help explain the reported meaningful reduction of post-void residual urine volume (PVR) in detrusor underactivity (DU) patients after thrice daily oral intake of acotiamide 100 mg for 2 weeks. METHODS: Effect of acotiamide [1–16μM] was assessed on nerve-mediated contractions evoked by electrical field stimulation (EFS) for 5s with 5ms pulse trains of 10V in longitudinal, mucosa intact rat and human bladder strips to construct frequency response curve (1–32Hz) and repeat 10Hz stimulation at 60s interval. Effect of acotiamide 2μM on spontaneous and carbachol evoked contractions was also assessed. RESULTS: Acotiamide 2μM significantly enhanced the Atropine and Tetrodotoxin (TTX)-sensitive EFS evoked contractions of rat and human bladder at 8–32Hz (Two-way ANOVA followed Sidak’s multiple comparison; *p

Aims To present a synopsis of the presentations and discussions from Think Tank I, “Implications for afferent–urothelial bidirectional communication” of the 2014 International Consultation on Incontinence-Research Society (ICI-RS) meeting in Bristol, UK.

Aims: To determine the role of p75 neurotrophin receptor (p75NTR) and the therapeutic effect of the selective small molecule p75NTR modulator, LM11A-31, in spinal cord injury (SCI) induced lower urinary tract dysfunction (LTUD) using a mouse model. Methods: Adult female T8-T9 transected mice were gavaged daily with LM11A-31 (100mg/kg) for up to 6 weeks, starting 1 day before, or 7 days following injury. Mice were evaluated in vivo using urine spot analysis, cystometrograms (CMGs), and external urethral sphincter (EUS) electromyograms (EMGs); and in vitro using histology, immunohistochemistry, and Western blot. Results: Our studies confirm highest expression of p75NTRs in the detrusor layer of the mouse bladder and lamina II region of the dorsal horn of the lumbar-sacral (L6-S1) spinal cord which significantly decreased following SCI. LM11A-31 prevented or ameliorated the detrusor sphincter dyssynergia (DSD) and detrusor overactivity (DO) in SCI mice, significantly improving bladder compliance. Furthermore, LM11A-31 treatment blocked the SCI-related urothelial damage and bladder wall remodeling. Conclusion: Drugs targeting p75NTRs can moderate DSD and DO in SCI mice, may identify pathophysiological mechanisms, and have therapeutic potential in SCI patients.

While urothelial signals, including sonic hedgehog (Shh), drive bladder mesenchyme differentiation, it is unclear which pathways within the mesenchyme are critical for its development. Studies have shown that fibroblast growth factor receptor (Fgfr)2 is necessary for kidney and ureter mesenchymal development. The objective of the present study was to determine the role of Fgfr2 in the bladder mesenchyme. We used Tbx18cre mice to delete Fgfr2 in the bladder mesenchyme ( Fgfr2 BM−/−). We performed three-dimensional reconstructions, quantitative real-time PCR, in situ hybridization, immunolabeling, ELISAs, immunoblot analysis, void stain on paper, ex vivo bladder sheet assays, and in vivo decerebrated cystometry. Compared with control bladders, embryonic day 16.5 (E16.5) Fgfr2 BM−/− bladders had thin muscle layers with less α-smooth muscle actin and thickened lamina propria with increased collagen type Ia and IIIa that intruded into the muscle. The reciprocal changes in mutant layer thicknesses appeared partly due to a cell fate switch. From postnatal days 1 to 30, Fgfr2 BM−/− bladders demonstrated progressive muscle loss and increased collagen expression. Postnatal Fgfr2 BM−/− bladder sheets exhibited decreased agonist-mediated contractility and increased passive stretch tension versus control bladder sheets. Cystometry revealed high baseline and threshold pressures and shortened intercontractile intervals in Fgfr2 BM−/− versus control bladders. Mechanistically, whereas Shh expression appeared normal, mRNA and protein readouts of hedgehog activity were increased in E16.5 Fgfr2 BM−/− versus control bladders. Moreover, E16.5 Fgfr2 BM−/− bladders exhibited higher levels of Cdo and Boc, hedgehog coreceptors that enhance sensitivity to Shh, compared with control bladders. In conclusion, loss of Fgfr2 in the bladder mesenchyme leads to abnormal bladder morphology and decreased compliance and contractility.

The isolated bladder shows autonomous micromotions, which increase with bladder distension, generate sensory nerve activity, and are altered in models of urinary dysfunction. Intravesical pressure resulting from autonomous activity putatively reflects three key variables; the extent of micromotion initiation, distances over which micromotions propagate, and overall bladder tone. In vivo, these variables are subordinate to the efferent drive of the central nervous system. In the micturition cycle storage phase, efferent inhibition keeps autonomous activity generally at a low level, where it may signal 'state of fullness', whilst maintaining compliance. In the voiding phase, mass efferent excitation elicits generalised contraction (global motility initiation). In lower urinary tract dysfunction, efferent control of the bladder can be impaired, for example due to peripheral 'patchy' denervation. In this case, loss of efferent inhibition may enable unregulated micromotility, and afferent stimulation, predisposing to urinary urgency. If denervation is relatively slight, the detrimental impact on voiding may be low, as the adjacent innervated areas may be able to initiate micromotility synchronous with the efferent nerve drive, so that even denervated areas can contribute to the voiding contraction. This would become increasingly inefficient the more severe the denervation, such that ability of triggered micromotility to propagate sufficiently to engage the denervated areas in voiding declines, so the voiding contraction increasingly develops the characteristics of underactivity. In summary, reduced peripheral coverage by the dual efferent innervation (inhibitory and excitatory) impairs regulation of micromotility initiation and propagation, potentially allowing emergence of overactive bladder and, with progression, detrusor underactivity.

Introduction Neural stimulation has become an established minimally invasive treatment for various lower urinary tract symptoms. The results both short- and long-term are encouraging, however, there is still a lack of knowledge of obvious risk factors, which may affect the outcome of treatment. Although neural stimulation has been embraced by healthcare professionals and patients, the exact mechanism by which neural stimulation works is still unclear. Discussion A condense review of knowledge available on this topic is presented. Several research questions are raised. Outlines of research studies, both clinical and basic science, are suggested. Conclusions Further studies are necessary to understand mechanism of action of neural stimulation and its implications on treatment outcomes. Neurourol. Urodynam. 33:618–621, 2014. © 2014 Wiley Periodicals, Inc.

Aims To present a brief review on discussions from “Do we understand any more about lower urinary tract interstitial cells?” session at the 2013 International Consultation on Incontinence-Research Society (ICI-RS) meeting in Bristol, UK. Methods Discussion focused on bladder interstitial cell (IC) subtypes, their localization and characterization, and communication between themselves, the urothelium, and detrusor smooth muscle. The role of ICs in bladder pathologies and new methods for studying ICs were also addressed. Results ICs have been studied extensively in the lower urinary tract and have been characterized based on comparisons with ICs of Cajal in the gastro-intestinal tract. In fetal bladders it is believed that ICs drive intrinsic contractions to expel urine through the urachus. These contractions diminish postpartum as bladder innervation develops. Voiding in human neonates occurs when filling triggers a spinal cord reflex that contracts the detrusor; in rodents, maternal stimulation of the perineum triggers voiding. Following spinal cord injury, intrinsic contractions, and spinal micturition reflexes develop, similar to those seen during neonatal development. These enhanced contractions may stimulate nociceptive and mechanosensitive afferents contributing to neurogenic detrusor overactivity and incontinence. The IC-mediated activity is believed to be initiated in the lamina propria by responding to urothelial factors. These IC may act syncytially through gap junction coupling and modulate detrusor activity through unknown mechanisms. Conclusion There has been a great deal of information discovered regarding bladder ICs, however, many of their (patho)physiological functions and mechanisms are still unclear and necessitates further research. Neurourol. Urodynam. 33:573–576, 2014. © 2014 Wiley Periodicals, Inc.

Background Botulinum neurotoxin A (BoNTA), which alleviates overactive bladder symptoms, is thought to act predominantly via the inhibition of transmitter release from parasympathetic nerves. However, actions at other sites such as afferent nerve terminals are possible. Objective To evaluate the effects of BoNTA on bladder afferent neuropeptide release and firing. Design, setting, and participants One side of the bladder of control and chronic (1–2 wk) spinal cord transected (SCT; T 8 -T 9 ) adult female mice was injected with BoNTA (0.5 U/5μl saline). After 48h, bladders with L 6 -S 2 spinal nerves were prepared for in vitro recordings. Outcome measurements and statistical analysis In bladder preparations, tension and optical mapping of Ca 2+ transients were used to measure intrinsic contractions, those evoked by capsaicin or the electrical stimulation of spinal nerves. Afferent firing was evoked by stretch or intrinsic bladder contractions. The numbers of responding units and firing rates were measured. Animal numbers were used to detect moderate to large between-group differences based on Cohen's criteria. Two-way analysis of variance was used to test spatial/temporal differences in Ca 2+ signals as mean plus or minus standard deviation. Differences between data sets were tested with the student t test and skewed data sets with a Mann-Whitney U test (significant when p Results and limitations In control and SCT bladders, BoNTA treatment decreased the contractions evoked by electrical stimulation of spinal nerves without altering intrinsic contractions. Afferent firing on untreated sides in response to stretch/intrinsic contractions was increased in SCTs versus controls. On BoNTA-treated sides, afferent firing rates were greatly attenuated in response to mechanical stimulation as were the capsaicin-evoked optical signals mediated by neuropeptide release. Conclusions SCT caused an increased sensitivity of afferent nerves to mechanical stimulation that was reduced by BoNTA treatment. Increased intrinsic activity after SCT was unaffected by the toxin. Thus BoNTA suppresses neurogenic detrusor overactivity by targeting afferent as well as efferent pathways in the bladder.

To measure the relative transcription of adenosine receptor subtypes and the contractile effects of adenosine and selective receptor-subtype ligands on detrusor smooth muscle from patients with neuropathic overactive (NDO) and stable bladders and also from guinea-pigs. Contractile function was measured at 37°C in vitro from detrusor smooth muscle strips. Contractions were elicited by superfusate agonists or by electrical field stimulation. Adenosine-receptor (A1, A2A, A2B, A3) transcription was measured by RT-PCR. Adenosine attenuated nerve-mediated responses with equivalent efficacy in human and guinea-pig tissue (pIC50 3.65–3.86); the action was more effective at low (1–8 Hz) compared to high (20–40 Hz) stimulation frequencies in human NDO and guinea-pig tissue. With guinea-pig detrusor the action of adenosine was mirrored by the A1/A2-agonist N-ethylcarboxamidoadenosine (NECA), partly abolished in turn by the A2B-selectve antagonist alloxazine, as well as the A1-selective agonist N6- cyclopentyladenosine (CPA). With detrusor from stable human bladders the effects of NECA and CPA were much smaller than that of adenosine. Adenosine also attenuated carbachol contractures, but mirrored by NECA (in turn blocked by alloxazine) only in guinea-pig tissue. Adenosine receptor subtype transcription was measured in human detrusor and was similar in both groups, except reduced A2A levels in overactive bladder. Suppression of the carbachol contracture in human detrusor is independent of A-receptor activation, in contrast to an A2B-dependent action with guinea-pig tissue. Adenosine also reduced nerve-mediated contractions, by an A1- dependent action suppressing ATP neurotransmitter action. Electronic supplementary material The online version of this article (doi:10.1007/s00210-016-1255-1) contains supplementary material, which is available to authorized users.

Spinal cord transection (SCT) leads to an increase in spontaneous contractile activity in the isolated bladder that is reminiscent of an overactive bladder syndrome in patients with similar damage to the central nervous system. An increase in interstitial cell number in the suburothelial space between the urothelium and detrusor smooth muscle layer occurs in SCT bladders, and these cells elicit excitatory responses to purines and pyrimidines such as ATP, ADP, and UTP. We have investigated the hypothesis that these agents underlie the increase in spontaneous activity. Rats underwent lower thoracic spinal cord transection, and their bladder sheets or strips, with intact mucosa except where specified, were used for experiments. Isometric tension was recorded and propagating Ca2+ and membrane potential ( Em) waves were recorded by fluorescence imaging using photodiode arrays. SCT bladders were associated with regular spontaneous contractions (2.9 ± 0.4/min); ADP, UTP, and UDP augmented the amplitude but not their frequency. With strips from such bladders, a P2Y6-selective agonist (PSB0474) exerted similar effects. Fluorescence imaging of bladder sheets showed that ADP or UTP increased the conduction velocity of Ca2+/ Em waves that were confined to regions of the bladder wall with an intact mucosa. When transverse bladder sections were used, Ca2+/ Em waves originated in the suburothelial space and propagated to the detrusor and urothelium. Analysis of wave propagation showed that the suburothelial space exhibited properties of an electrical syncitium. These experiments are consistent with the hypothesis that P2Y-receptor agonists increase spontaneous contractile activity by augmenting functional activity of the cellular syncitium in the suburothelial space.

Background Botulinum neurotoxins type A (BoNT/A), β3-adrenergic receptor agonists, and phosphodiesterase type 5 (PDE5) inhibitors are promising agents that mitigate lower urinary tract symptoms by attenuating the sensory system. However, whether they act directly on afferent nerves or indirectly through the other cell types is unclear. Methods Spinal cord transected female mice were used as a model for neurogenic bladder overactivity. In vivo methods utilized decerebrate mouse cystometry. In vitro approaches included optical mapping of Ca2+ transient, single unit afferent nerve recordings and tension measurements from bladder sheets and wall cross-sections. Immunohistochemistry was used to measure the expression of β3-adrenergic receptors on dorsal root ganglion (DRG) neurons. Results Our unique approaches revealed the direct effects of BoNT/A in inhibiting neuropeptide release and firing rates in afferents following bladder injections. β3-adrenergic receptor agonists are demonstrated to directly inhibit afferent nerve firing independent of the relaxing effects on bladder smooth muscle. Moreover, data suggest the expression of these receptors on DRG neurons that send projections to the bladder. The mechanism of action of PDE5 inhibitors on bladder overactivity is discussed. Discussion The questions raised during the plenary session of the 2011 International Consultation on Incontinence-Research Society meeting regarding the benefits of BoNT/A, β3-adrenergic receptor agonist and PDE5 inhibitor treatments of overactive bladder are addressed. Conclusion Our findings suggest that the abovementioned agents, in low enough concentrations, can directly inhibit afferent excitability without decreasing detrusor contractility. Accordingly, they have considerable potential for treating the sensory component of lower urinary tract dysfunctions. Neurourol. Urodynam. 31:300–308, 2012. © 2012 Wiley Periodicals, Inc.

Common methods for inducing neurogenic bladder overactivity include: (1) spinal cord transection or contusion injury, (2) supraspinal injury (decerebration, local lesions, middle cerebral artery occlusion and (3) systemic (e.g., cyclophosphamide) or intravesical administration of irritant or inflammatory agents (e.g., acrolein, acid, lipopolysaccharide). These insults can cause peripheral, spinal and supraspinal effects which are difficult to resolve using traditional cystometry alone. To address this problem, we have developed new methods as well as new highly reproducible models to study neurogenic bladder dysfunction in the mouse. These models include direct sensitization of the bladder using ionizing radiation or indirect sensitization through colonic irradiation (figure 1). The classic model for neurogenic bladder overactivity is spinal cord injury which results in non-voiding contractions during the filling phase. However, there is also an intrinsic myogenic component that is present when studied in vitro. Our radiation models are purely neurogenic and do exhibit overactivity in isolated tissues. Figure: 1 New in vitro studies utilize bladder-urethra sheets, spinal cord slices and in-line cultured cells. Bladder-urethra sheets can be isolated with the hypogastric (T11-L2) and pelvic (L6-S2) nerves and imaged for the propagation of optical activity along the bladder wall simultaneously with afferent nerve and tension recordings. Alternatively, bladder sheets can be mounted vertically as cross-sections for measuring the spread of activity across the wall from mucosa to serosa. Spinal cord slices are sectioned 0.5 to 1 mm thick to include the dorsal and ventral roots which can be stimulated to optically map pathologically-induced changes in spinal cord circuitry. In-line cultured cells, on the other hand, allow for different linear arrangements of urothelial, interstitial, smooth muscle and dorsal root ganglion cells to map alterations in the flow of information in cell-to-cell communication. In vivo studies utilize cystometry which can be combined with cerebro-cortical imaging of optical activity during bladder filling and emptying. The new models of neurogenic overactivity are induced by irradiation of the bladder (direct sensitization) or colon (indirect cross-sensitization) both of which result in inflammation and urothelial damage leading to bladder overactivity. The changes induced by these models have been compared to those induced by the well established models of chronic spinal cord transection at the thoracic (T8-T9) and lumbar (L4-L5) levels.

A substantial portion of the current research on lower urinary tract dysfunction is focused on afferent mechanisms. The main goals are to define and modulate the signaling pathways by which afferent information is generated, enhanced and conveyed to the central nervous system. Alterations in bladder afferent mechanisms are a potential source of voiding dysfunction and an emerging source for drug targets. Established drug therapies such as muscarinic receptor antagonists, and two emerging therapies, beta(3)-adrenergic receptor agonists and botulinum toxin type-A, may act partly through afferent mechanisms. This review focuses on these two new principles and new and established methods for determining their sites of action. It also provides brief information on the innervation of the bladder, afferent receptors and transmitters and how these may communicate with the urothelium, interstitial cells and detrusor smooth muscle to regulate micturition. Peripheral and central mechanisms of afferent sensitization and myogenic mechanisms that lead to detrusor overactivity, overactive bladder symptoms and urgency sensations are also covered. This work is the result from 'Think Tank' presentations, and the lengthy discussions that followed, at the 2010 International Consultation on Incontinence Research Society meeting in Bristol, UK. Neurourol. Urodynam. 30:684-691, 2011.

The joy of eating is one that people wish to enjoy throughout their lives, and the loss of the ability to orally ingest food reduces QOL. Therefore, nutritional management for facility residents must aim to ensure an appropriate energy intake for each individual.The measures we implemented after conducting a survey on food texture at welfare facilities for the elderly in Okayama city in September 2007 produced favorable results. In the present study, we report the positive results obtained from a follow- up survey conducted half a year later in April 2008.Subjects were residents at welfare facilities for the elderly, specifically healthcare and welfare facilities for elderly requiring long-term care. Individuals receiving tube feeding were excluded.Survey items included confirmation of consistency using a food texture checklist, care requirement level, level of independence in activities of daily living for disabled or elderly individuals, walking ability, oral function, presence or absence of dentures, dentition, and BMI.Survey results confirmed that care requirement level was correlated with the degree of bedridden state, walking ability, and oral function, and that it also influenced food texture. In particular, the selection of food texture was influenced by factors such as oral function, presence or absence of dentures, and dentition. Moreover, introduction of soft food which solidified paste food with some coagulants enabled almost all subjects to maintain or increase their BMI over the six-month period.These findings indicate that registered dietitians must monitor not only food consumption, but also care requirement level, degree of bedridden state, walking ability, presence or absence of dentures, and dentition.The present support system enabled the provision of safe and satisfying meals to facility residents and was thought to have contributed greatly to care for maintaining the dignity of elderly individuals from the perspective of nutritional management.

Increased gap junction expression in lamina propria myofibroblasts and urothelial cells may be involved in detrusor overactivity, leading to incontinence. Immunohistochemistry was used to compare connexin (Cx) 26, 43, and 45 expression in the bladders of neonatal, adult, and spinal cord-transected rats, while optical imaging was used to map the spread of spontaneous activity and the effects of gap junction blockade. Female adult Sprague-Dawley rats were deeply anesthetized, a laminectomy was performed, and the spinal cord was transected (T8/T9). After 14 days, their bladders and those of age-matched adults (4 mo old) and neonates (7–21 day old) were excised and studied immunohistochemically using frozen sections or optically using whole bladders stained with voltage- and Ca2+-sensitive dyes. The expression of Cx26 was localized to the urothelium, Cx43 to the lamina propria myofibroblasts, and Cx45 to the detrusor smooth muscle. While the expression of Cx45 was comparable in all bladders, the expression of Cx43 and Cx26 was increased in neonate and transected animals. In the bladders of adults, spontaneous activity was initiated at multiple sites, resulting in a lack of coordination. Alternatively, in neonate and transected animals spontaneous activity was initiated at a focal site near the dome and spread in a coordinated fashion throughout the bladder. Gap junction blockade (18β-glycyrrhetinic acid, 1 μM) abolished this coordinated activity but had no effect on the uncoordinated activity in adult bladders. These data suggest that coordinated spontaneous activity requires gap junction upregulation in urothelial cells and lamina propria myofibroblasts.

OBJECTIVE: To assess the safety and efficacy of anticholinergics in men with lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia (BPH) by a systematic review of published reports and a meta-analysis of the reported outcomes. METHODS: We searched Medline, Embase and Cochrane databases (1966-2006), and hand-searched relevant reference lists and conference proceedings, for studies on the use of anticholinergics in men with BPH or bladder outlet obstruction. Eligible studies were assessed for quality and foreign language studies were translated. We collected data on all reported outcomes, conducted meta- analyses on the maximum urinary flow rate (Q(max)), postvoid residual urine volume (PVR) and volume at first contraction, and calculated the acute urinary retention (AUR) rate. We used sensitivity analysis to confirm the findings. RESULTS: We identified five randomized controlled trials (RCTs) and 15 observational studies. Four RCTs incorporating 633 patients were included in the meta-analyses. Anticholinergics did not significantly alter Q(max) (0.1 mL/s, 95% confidence interval, CI, 0.6-0.7). The PVR was increased by 11.6 mL (95% CI 4.5-18.6) although there was no significant difference between AUR rates. The total International Prostate Symptom Scores (IPSS) were not significantly different, but there were improvements for IPSS storage subscores in one RCT. The AUR rate was 0.3% at the 12-week follow-up in 365 men in the RCTs and observational studies. CONCLUSION: Anticholinergic use in men with LUTS suggestive of BPH appears to be safe. Further studies are required to establish efficacy with a suitable precision.

INTRODUCTION AND OBJECTIVES: Many urologic conditions are associated with dysfunction of autonomic bladder innervation. A precise understanding of the anatomical relationship of the bladder neck and its autonomic nervous system could substantially improve treatment modalities. Currently there is no comprehensive model that provides a detailed three-dimensional (3D) view of bladder neck innervation. Therefore, we sought to create a 3D reconstruction of autonomic nervous tissue innervating the bladder neck using male and female cadaver histopathology. METHODS: We obtained intact pelvic tissues from a male and female cadaver. Axial cross-sections of the bladder neck were generated at 3e5mm intervals and stained for S100 protein. Distances between autonomic nerves and bladder mucosa were recorded. Nerve tracings were manually demarcated and imported into SolidWorks (Waltham, MA, USA) and Blender (Amsterdam, Netherlands) software programs to generate 3D reconstructions of autonomic nerve anatomy. RESULTS: Longitudinal and circular nerve tracings were successfully demarcated and converted into a 3D image reconstruction. We successfully precisely characterized anatomic nerve distributions. Axial cross-sections and 3D images showed that autonomic innervation was highly concentrated in the posterior aspect of the bladder neck in both male and female bladder specimens (Figures 1 and 2). The mean distances between autonomic nerve tissue and the bladder mucosa was 1.15mm posteriorly and 4.0mm anteriorly in the male bladder (0.27-2.87 vs. 2.03-6.20, p

The micturition reflex involves afferent nerve activation when the bladder is sufficiently full and subsequent controlled firing of parasympathetic efferent nerves to contract the detrusor muscle as part of the voiding mechanism. Alteration of the sensitivity of afferent activation or loss of control over transmitter release could lead to sensory- or motor-activated incontinence, respectively. The control mechanisms that regulate these 2 activities remain poorly understood. Current opinion is that the sensation of bladder fullness is relayed by afferent nerves in the mucosal layer, which are activated by the release of chemical mediators, such as adenosine triphosphate (ATP), from the urothelium when it is stretched as the bladder fills. This hypothesis supports the concept that other chemical signals that affect bladder sensation (eg, changes in urine composition and agents such as capsaicin) can modulate the sensitivity of the basic system. It has also been proposed that a layer of myofibroblasts immediately below the basal lamina of the urothelium acts as a variable gain regulator of the sensory process between ATP release and afferent excitation. These myofibroblasts are functionally connected to form an electrical syncytium, make close contact with nerves, and respond by generating electrical responses and transient increases in intracellular Ca2+ when exposed to ATP. On the efferent side, using a guinea pig detrusor model, possible modulators of transmitter release have been investigated, including adenosine (the breakdown product of the neurotransmitter ATP). Adenosine reduces the force of nerve-mediated contractions by acting predominantly at presynaptic sites at the nerve-muscle junction via a subtype of an adenosine receptor-the A1 receptor. An additional effect, possibly via A2 receptors, is also present on the detrusor muscle itself. These actions of adenosine are less evident in human detrusor muscle but remain a potential modulatory target. In summary, the cellular and molecular regulation of bladder fullness sensation and efferent transmitter release are becoming better understood and represent potential drug targets for the management of detrusor overactivity.

Contractile activation of detrusor smooth muscle is initiated by the release of transmitters from motor nerves. Acetylcholine is a ubiquitous transmitter, as also is adenosine triphosphate (ATP) in many animal bladders and in people from several patient groups with pathological bladder function. In recent years there has been progress in explaining several cellular mechanisms that link transmitter release to contraction and these will be considered. The lifetime of ATP in the neuromuscular junction is finite and broken down ultimately to adenosine, which can exert modulatory control of contractile activation. Adenosine depresses nerve-mediated contractions and two sites of action have been proposed: an action on the motor nerves via A receptors to depress further transmitter release and a less well-defined depressant effect on the detrusor muscle. The Ca2+ ions that activate the contractile proteins are derived from intracellular stores, which releases their content via IP receptor activation and Ca2+-induced Ca2+ release. Filling of the stores in the rest interval is mediated via transmembrane flux of Ca2+through Ca2+ channels. Activation of the channels is regulated by the level of the intracellular [Ca2+], via activation and inactivation of Ca2+-sensitive K channels. Thus, Ca2+ store filling is regulated by intracellular [Ca2+] via a negative feedback process. The presence and physiological function of spontaneous contractions in detrusor remain contentious and little is known about their origin. One possibility is that they originate from random Ca2+ sparks, i.e. localized transient increases of [Ca2+] that may eventually progress to generate a cellular Ca2+ transient. Observations by confocal microscopy have revealed the presence of such sparks, especially near the cell membrane, and thus provide a cellular basis for spontaneous contractions. Finally, the questions arises as to whether detrusor smooth muscle is a functional syncitium. The demonstration of small gap junctions by electron microscopy and the demonstration of the gap junction protein connexin45 indicate that the muscle mass may indeed be functionally connected. The implications regarding the spread of excitation are discussed.

Neural stimulation has become an established minimally invasive treatment for various lower urinary tract symptoms. The results both short- and long-term are encouraging, however, there is still a lack of knowledge of obvious risk factors, which may affect the outcome of treatment. Although neural stimulation has been embraced by healthcare professionals and patients, the exact mechanism by which neural stimulation works is still unclear.A condense review of knowledge available on this topic is presented. Several research questions are raised. Outlines of research studies, both clinical and basic science, are suggested.Further studies are necessary to understand mechanism of action of neural stimulation and its implications on treatment outcomes.

To present a brief review on discussions from "Do we understand any more about lower urinary tract interstitial cells?" session at the 2013 International Consultation on Incontinence-Research Society (ICI-RS) meeting in Bristol, UK.Discussion focused on bladder interstitial cell (IC) subtypes, their localization and characterization, and communication between themselves, the urothelium, and detrusor smooth muscle. The role of ICs in bladder pathologies and new methods for studying ICs were also addressed.ICs have been studied extensively in the lower urinary tract and have been characterized based on comparisons with ICs of Cajal in the gastro-intestinal tract. In fetal bladders it is believed that ICs drive intrinsic contractions to expel urine through the urachus. These contractions diminish postpartum as bladder innervation develops. Voiding in human neonates occurs when filling triggers a spinal cord reflex that contracts the detrusor; in rodents, maternal stimulation of the perineum triggers voiding. Following spinal cord injury, intrinsic contractions, and spinal micturition reflexes develop, similar to those seen during neonatal development. These enhanced contractions may stimulate nociceptive and mechanosensitive afferents contributing to neurogenic detrusor overactivity and incontinence. The IC-mediated activity is believed to be initiated in the lamina propria by responding to urothelial factors. These IC may act syncytially through gap junction coupling and modulate detrusor activity through unknown mechanisms.There has been a great deal of information discovered regarding bladder ICs, however, many of their (patho)physiological functions and mechanisms are still unclear and necessitates further research. Neurourol. Urodynam. 33:573-576, 2014. © 2014 Wiley Periodicals, Inc.

Mandibular movements are coordinated ones caused by TMJ, muscles, dentitions and neuromuscles in stomatognathic system. The pathways in areas of intercuspal position (ICP) during chewing movements which are particularly influenced by dentitions were investigated. An evaluation of these movements will contribute to establising a functional diagnosis method.An objective assessment fitting the regression line to sample data on pathways in areas of ICP during chewing movements was also reported. In addition to these studies, we developed a system to measurement of mandibular movements during grinding movement utilizing visual feedback system.In this study, it became possible to establish a new displaying method for tooth grinding curved surfaces and pathways during chewing movements by applying algorithms of perspective transformation, hidden line elimination and coordinate transformation to incisal 3-dimensional data (X, Y, Z) obtained from a Sirognathograph (SGG).The results were as follows.:1. Tooth grinding curved surfaces were constructed as the wire frame model.2. Pathways in areas of ICP during chewing movements were displayed on the tooth grinding curved surfaces.3. A new method makes possible to display the relation between tooth grinding curved surfaces as anatomical information and pathways in areas of ICP chewing movements as functional information.

In 1993,portraits photographed by Toyo KIKUCHI were found from the KUKUCHI family's KURA (Japanese traditional warehouse). The number of the portraits is more than forty, and two albums with thirty portraits were also found. In New York, Toyo KIKUCHI had successed his new buisiness namely "Home Portrait" during the late MEIJI and the early TAISHOU eras. These photographs (8×10 inches) with brilliant highlight and deep shadow tones in sepia color were taken by him at his studio in New York. Toyo's photographs, which were found last year, were evaluated by the current leading photographers as the most excellent studio portraits not only at that time but also with the modern sense of photographic art. On the other hand, Toyo mastered photographic emulsion technology in U.S.A., and he established Oriental Photo Industrial Co., Ltd., a wellknown manufacuturer of photographic materials, after his return from New York. Hitherto, Toyo KIKUCHI's achievemwnts were known as the enterpriser and also the emulsion engineer in photographic industry rather than the studio photographer. The photographs discovered, however, are the definite evidence of his quite outstanding skills in studio photography. The authors worked out "The Exhibition of Home Portrait by Toyo KIKUCHI" which held during June and July, 1994 at THE SHADAI GALLERY. A part of this paper was prepared for the material of the exhibition and deals with the genealogical survey of Toyo KIKUCHI as a studio photographer and also an emulsion engineer.

症例は63歳,女性,著明な左胸水と縦隔腫瘤にて発症.胸水腫瘍細胞の表面抗原の検索でCD56, CD7, CD33陽性, CD3, CD16, CD57陰性にて, Myeloid/Natural Killer(以下NK)前駆細胞性急性白血病と診断した.骨髄中には腫瘍細胞を認めなかった. JALSGAML97プロトコールにより化学療法を施行し,完全寛解に至った.貯留胸水細胞より診断し得た本症は文献的に初めてであり報告した.

Botulinum neurotoxins type A (BoNT/A), β(3)-adrenergic receptor agonists, and phosphodiesterase type 5 (PDE5) inhibitors are promising agents that mitigate lower urinary tract symptoms by attenuating the sensory system. However, whether they act directly on afferent nerves or indirectly through the other cell types is unclear.Spinal cord transected female mice were used as a model for neurogenic bladder overactivity. In vivo methods utilized decerebrate mouse cystometry. In vitro approaches included optical mapping of Ca(2+) transient, single unit afferent nerve recordings and tension measurements from bladder sheets and wall cross-sections. Immunohistochemistry was used to measure the expression of β(3)-adrenergic receptors on dorsal root ganglion (DRG) neurons.Our unique approaches revealed the direct effects of BoNT/A in inhibiting neuropeptide release and firing rates in afferents following bladder injections. β(3)-adrenergic receptor agonists are demonstrated to directly inhibit afferent nerve firing independent of the relaxing effects on bladder smooth muscle. Moreover, data suggest the expression of these receptors on DRG neurons that send projections to the bladder. The mechanism of action of PDE5 inhibitors on bladder overactivity is discussed.The questions raised during the plenary session of the 2011 International Consultation on Incontinence-Research Society meeting regarding the benefits of BoNT/A, β(3)-adrenergic receptor agonist and PDE5 inhibitor treatments of overactive bladder are addressed.Our findings suggest that the abovementioned agents, in low enough concentrations, can directly inhibit afferent excitability without decreasing detrusor contractility. Accordingly, they have considerable potential for treating the sensory component of lower urinary tract dysfunctions.

The addition of 1-chloroalkyl p-tolyl sulfoxides to carbonyl compounds gave adducts which were then converted to chlorooxiranes in two steps with good overall yields. The treatment of the chlorooxiranes with various nucleophiles gave α-hygroxy α′-substituted ketones or α-hydroxy ketones in good yields.