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Functional gastrointestinal disorders (FGIDs) have prominent sex differences in incidence, symptoms, and treatment response that are not well understood. Androgens are steroid hormones present at much higher levels in males than females and could be involved in these differences. In adults with irritable bowel syndrome (IBS), a FGID that affects 5% to 10% of the population worldwide, we found that free testosterone levels were lower than those in healthy controls and inversely correlated with symptom severity. To determine how this diminished androgen signaling could contribute to bowel dysfunction, we depleted gonadal androgens in adult mice and found that this caused a profound deficit in gastrointestinal transit. Restoring a single androgen hormone was sufficient to rescue this deficit, suggesting that circulating androgens are essential for normal bowel motility in vivo. To determine the site of action, we probed androgen receptor expression in the intestine and discovered, unexpectedly, that a large subset of enteric neurons became androgen-responsive upon puberty. Androgen signaling to these neurons was required for normal colonic motility in adult mice. Taken together, these observations establish a role for gonadal androgens in the neural regulation of bowel function and link altered androgen levels with a common digestive disorder., Introduction Almost all disorders of the gut-brain axis, from inflammatory bowel disease to neurological disorders with GI comorbidities, exhibit sex differences (1-3). Irritable bowel syndrome (IBS) is one of the [...]

Background Currently available tocolytic agents are not effective treatment for preterm labor beyond 48 h. A major reason is the development of maternal side effects which preclude the maintenance of an effective steady-state drug concentration. One strategy that can mitigate these side effects is utilizing synergistic drug combinations to reduce the drug concentrations necessary to elicit a clinical effect. We have previously shown that three anoctamin 1 (ANO1) antagonists mediate potent relaxation of precontracted human uterine smooth muscle (USM). In this study, we aimed to determine whether a combination of sub-relaxatory doses of tocolytic drugs in current clinical use [the L-type voltage-gated calcium channel (VGCC) blocker, nifedipine (NIF); and the [beta].sub.2-adrenergic ([beta]2AR) agonist, terbutaline (TRB)] will potentiate USM relaxation with two ANO1 antagonists [benzbromarone (BB) and MONNA (MN)]. Objective This study sought to examine the synergistic potency and mechanistic basis of two ANO1 antagonists with currently available tocolytic drugs. Functional endpoints assessed included relaxation of pre-contracting pregnant human USM tissue, inhibition of intracellular calcium release, and reduction of spontaneous transient inward current (STIC) recordings in human uterine smooth muscle cells. Methods Human myometrial strips and primary human USM cells were used in organ bath and calcium flux experiments with different combinations of sub-threshold doses of ANO1 antagonists and terbutaline or nifedipine to determine if ANO1 antagonists potentiate tocolytic drugs. Results The combination of sub-threshold doses of two ANO1 antagonists and current tocolytic drugs demonstrate a significant degree of synergy to relax human pregnant USM compared to the effects achieved when these drugs are administered individually. Conclusion A combination of sub-threshold doses of VGCC blocker and [beta]2AR agonist with ANO1 antagonists potentiates relaxation of oxytocin-induced contractility and calcium flux in human USM ex vivo. Our findings may serve as a foundation for novel tocolytic drug combinations. Keywords: ANO1, Calcium-activated chloride channel, TMEM16A, Uterine smooth muscle, Tocolytics, Author(s): Shunsuke Hyuga[sup.1], Robert C. Parry[sup.1], Jennifer Danielsson[sup.1], Joy Vink[sup.2], Xiao Wen Fu[sup.1], Amy Wu[sup.1], William Dan[sup.1], Peter D. Yim[sup.1] and George Gallos[sup.1] Introduction Preterm birth (PTB) remains a major [...]

Introduction γ-amino butyric acid (GABA) is not only the major inhibitory neurotransmitter in the central nervous system (CNS), but it also plays an important role in the lung, mediating airway smooth muscle relaxation and mucus production. As kinases such as protein kinase A (PKA) are known to regulate the release and reuptake of GABA in the CNS by GABA transporters, we hypothesized that β-agonists would affect GABA release from airway epithelial cells through activation of PKA. Methods C57/BL6 mice received a pretreatment of a β-agonist or vehicle (PBS), followed by methacholine or PBS. Bronchoalveolar lavage (BAL) was collected and the amount of GABA was quantified using HPLC mass spectrometry. For in vitro studies, cultured BEAS-2B human airway epithelial cells were loaded with .sup.3H-GABA. .sup.3H-GABA released was measured during activation and inhibition of PKA and tyrosine kinase signaling pathways. Results β-agonist pretreatment prior to methacholine challenge attenuated in vivo GABA release in mouse BAL and .sup.3H-GABA release from depolarized BEAS-2B cells. GABA release was also decreased in BEAS-2B cells by increases in cAMP but not by Epac or tyrosine kinase activation. Conclusion β-agonists decrease GABA release from airway epithelium through the activation of cAMP and PKA. This has important therapeutic implications as β-agonists and GABA are important mediators of both mucus production and airway smooth muscle tone., Author(s): Jennifer Danielsson[sup.1] , Sarah Zaidi[sup.2] , Benjamin Kim[sup.3] , Hiromi Funayama[sup.1] [sup.4] , Peter D. Yim[sup.1] , Dingbang Xu[sup.1] , Tilla S. Worgall[sup.3] , George Gallos[sup.1] , Charles W. [...]

Tumor necrosis factor (TNF)-[alpha] is a potent inflammatory cytokine implicated in the exacerbation of asthma. Chronic exposure to TNF-[alpha] has been reported to induce G protein-coupled receptor desensitization, but adenylyl cyclase sensitization, in airway smooth muscle cells by an unknown mechanism. Cyclic AMP, which is synthesized by adenylyl cyclases in response to G protein-coupled receptor signals, is an important second messenger involved in the regulation of the airway muscle proliferation, migration, and tone. In other cell types, TNF-[alpha] receptors transactivate the EGF receptor, which activates raf-1 kinase. Further studies in transfected cells show that raf-1 kinase can phosphorylate and activate some isoforms of adenylyl cyclase. Cultured human airway smooth muscle cells were treated with TNF-[alpha] in the presence or absence of inhibitors of prostaglandin signaling, protein kinases, or Gi proteins. TNF-[alpha] caused a significant dose- (1-10 ng/ml) and time-dependent (24 and 48 h) increase in forskolin-stimulated adenylyl cyclase activity, which was abrogated by pretreatment with GW5074 (a raf-1 kinase inhibitor), was partially inhibited by an EGF receptor inhibitor, but was unaffected by pertussis toxin. TNF-[alpha] also increased phosphorylation of [Ser.sup.338] on raf-1 kinase, indicative of activation. IL-1[beta] and EGF sensitization of adenylyl cyclase activity was also sensitive to raf-1 kinase inhibition by GW5074. Taken together, these studies link two signaling pathways not previously characterized in human airway smooth muscle cells: TNF-[alpha] transactivation of the EGF receptor, with subsequent raf-1 kinase-mediated activation of adenylyl cyclase. GW5074; mitogen-activated protein kinase; phosphorylation; immunoblot; cell culture doi:10.1152/ajplung.00123.2006.

Vascular disease produces changes in lumenal shape evident in magnetic resonance angiography (MRA). However, quantification of vascular shape from MRA is problematic due to image artifacts. Prior deformable models for vascular surface reconstruction primarily resolve problems of initialization of the surface mesh. However, initialization can be obtained in a trivial manner for MRA using isosurfaces. We propose a methodology for deforming the isosurface to conform to the boundaries of objects in the image with minimal a priori assumptions of object shape. As in conventional methods, external forces attract the surface toward edges in the image. However, smoothing is produced by a moment that aligns the normals of adjacent surface triangles. Notably, the moment produces no translational motion of surface triangles. The deformable isosurface was applied to a digital phantom of a stenotic artery, to MRA of three renal arteries with atherosclerotic disease and MRA of one carotid artery with atherosclerotic disease. Results of the surface reconstruction from the deformable model were compared with conventional X-ray angiography for the renal arteries. Measurement of the degree of stenosis of the renal arteries was within 12% [+ or -] 6%. The deformable model provided improvements over the isosurface in all cases in terms of measurement of the degree of stenosis or improving the surface smoothness. Index Terms--Carotid artery, deformable model, magnetic resonance angiography, renal artery.

Three-dimensional (3-D) angiographic methods are gaining acceptance for evaluation of atherosclerotic disease. However, measurement of vessel stenosis from 3-D angiographic methods can be problematic due to limited image resolution and contrast. We present a method for reconstructing vessel surfaces from 3-D angiographic methods that allows for objective measurement of vessel stenosis. The method is a deformable model that employs a tubular coordinate system. Vertex merging is incorporated into the coordinate system to maintain even vertex spacing and to avoid problems of self-intersection of the surface. The deformable model was evaluated on clinical magnetic resonance (MR) images of the carotid (n = 6) and renal (n = 2) arteries, on an MR image of a physical vascular phantom and on a digital vascular phantom. Only one gross error occurred for all clinical images. All reconstructed surfaces had a realistic, smooth appearance. For all segments of the physical vascular phantom, vessel radii from the surface reconstruction had an error of less than 0.2 of the average voxel dimension. Variability of manual initialization of the deformable model had negligible effect on the measurement of the degree of stenosis of the digital vascular phantom. Index Terms--Atherosclerosis, deformable model, magnetic resonance angiography, surface reconstruction.

Interpretation of magnetic resonance angiography (MRA) is problematic due to complexities of vascular shape and to artifacts such as the partial volume effect. We present new methods to assist in the interpretation of MRA. These include methods for detection of vessel paths and for determination of branching patterns of vascular trees. They are based on the ordered region growing (ORG) algorithm that represents the image as an acyclic graph, which can be reduced to a skeleton by specifying vessel endpoints or by a pruning process. Ambiguities in the vessel branching due to vessel overlap are effectively resolved by heuristic methods that incorporate a priori knowledge of bifurcation spacing. Vessel paths are detected at interactive speeds on a 500-MHz processor using vessel endpoints. These methods apply best to smaller vessels where the image intensity peaks at the center of the lumen which, for the abdominal MRA, includes vessels whose diameter is less than 1 cm. Index Terms--Magnetic resonance angiography, skeletonization, visualization.