BackgroundIntestinal hyper-permeability plays a critical role in the etiopathogenesis of inflammatory bowel disease (IBD) by affecting the penetration of pathogens, toxic compounds and macromolecules. 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], the active form of vitamin D, has been shown to be an important regulator of IBD and recent epidemiology suggests that patients with IBD have an impaired vitamin D status. The purpose of this study is to investigate the possible protective effects of 1,25(OH)2D3 on mucosal injury and epithelial barrier disruption on dextran sulfate sodium (DSS)-induced acute colitis model.MethodsWe used DSS-induced acute colitis model to investigate the protective effects of 1,25(OH)2D3 on mucosal injury and epithelial barrier integrity. Severity of colitis was evaluated by disease activity index (DAI), body weight (BW) change, colon length, histology, myeloperoxidase (MPO) activity, and proinflammatory cytokine production including tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). In vitro the protective role of 1,25(OH)2D3 was assessed by incubating Caco-2 cells with or without DSS and measuring transepithelial electrical resistance (TEER) and fluorescein isothiocyanate dextran (FITC-D). The intestinal permeability was analyzed by FITC-D, bacterial translocation and measurement of lipopolysaccharide (LPS). Ultrastructural features of the colon tissue and Caco-2 cell monolayer were observed by electron microscopy. Expressions of tight junction (TJ) proteins in the colon mucosa and Caco-2 cells were detected by immunohistochemistry, immunofluorescence, Western blot and real-time fluorescent quantitative PCR, respectively.ResultsDSS-induced acute colitis model was characterized by a reduced BW, AUC of BW, serum calcium, higher DAI, AUC of DAI, shortened colon length, elevated MPO activity, worsened histologic inflammation, increased mononuclear cell numbers in mesenteric lymph nodes (MLNs) and colonic lamina propria (LP), and enhanced proteins and mRNA levels of TNF-α and IFN-γ. 1,25(OH)2D3 markedly increased expressions of TJ proteins and mRNA and decreased the FITC-D permeability and the level of LPS. Furthermore, 1,25(OH)2D3 abrogated bacterial translocation to MLNs and ameliorated ultrastructural features of the colon epithelium by scanning electron microscopy (SEM). In vitro, 1,25(OH)2D3 increased TEER, TJ proteins and mRNA expressions, decreased the FITC-D permeability, and preserved structural integrity of the TJ in Caco-2 cells.Conclusions1,25(OH)2D3 may play a protective role in mucosal barrier homeostasis by maintaining the integrity of junction complexes and in healing capacity of the colon epithelium. 1,25(OH)2D3 may represent an attractive and novel therapeutic agent for the adjuvant therapy of IBD.

The direct interaction of hyaluronic acid (HA) and proteins of the inter-␣-inhibitor family plays a critical role in organization and stabilization of the expanding cumulus extracellular matrix (cECM) following an ovulatory stimulus. Despite similarities in the morphology of cumulus oocyte complexes (COCs) expanding in vivo and in vitro, we find that the cECM of COCs which expand within intact follicles are more elastic and resistant to shear stress than the cECM of those stabilized in vitro. Western blot analysis shows that only the heavy chains of inter-␣-inhibitor are incorporated into the cECM and appears to be covalently linked to HA after stabilization in vivo while intact inter-␣-inhibitor is bound to the HA-enriched cECM by a non-covalent mechanism in in vitro stabilized COCs. However, purified pre-␣-inhibitor and HA can form covalent linkage in the presence of granulosa cells or with granulosa cellconditioned medium. In addition, COCs resistance to shear stress is also enhanced by coincubation with granulosa cells. Upon formation of the apparent covalent linkage between heavy chains and HA in culture medium, the light chain (bikunin) is concomitantly released into the medium as a complex with chondroitin sulfate moieties of inter-␣-inhibitor supporting the possibility that HA may replace the chondroitin sulfate linkage to the heavy chains. We speculate that a factor(s) secreted by granulosa cells within the follicle may catalyze a transesterification reaction resulting in an exchange of chondroitin sulfate with HA at the heavy chain/chondroitin sulfate junction followed by release of chondroitin sulfate-bikunin into the follicular fluid. It is also possible that the consequent further stabilization of the cECM through the covalent interaction of HA and heavy chains of inter-␣-inhibitor may play an important role in the process of ovulation.In most mammalian species (including mouse, rat, and human), cumulus-oocyte complexes (COCs) 1 of pre-ovulatory follicles undergo a dramatic change following an ovulatory stimulus. The tightly packed cumulus cells first disaggregate and then synthesize and secrete large amounts of hyaluronic acid (HA) into their extracellular matrices (ECMs). The ECM, cumulus cells, and oocyte are thus integrally bound within an expanded mucoid complex which is about 20 to 40 times larger (volume) dependent upon the species (1). This process of cumulus expansion is required for ovulation and may also facilitate the process of fertilization (2-4).We have previously identified a serum factor (proteins of the inter-␣-inhibitor family), critical in organizing and stabilizing the expanding cumulus matrix (5). This protein factor appears to be excluded from follicular fluid until the ovulatory gonadotropin surge and then quickly diffuses into the follicular fluid where it becomes integrated within the cumulus ECM (5, 6). Two major forms of this factor, pre-␣-inhibitor (P␣I) and inter-␣-inhibitor (I␣I), exist in mammalian species including mouse, bovine, and human (7,8). They each include a ...

Aerobic exercise could cause sympathetic nerve stimulation, consequently causing the expansion of the TM and SC, which, in turn, leads to IOP reduction. Furthermore, SC and the TM might have an autonomic regulation function, and their expansion and collapse might not be completely dependent on the IOP.

A relatively simple technique was used to estimate the size of the combined pools of zinc with which plasma zinc exchanges within 2 d (EZP). EZP size was determined from the amount of isotope introduced into the plasma and the coefficient of the simple exponential decay function fitting enrichment data between d 3 and 9 after isotope administration. Using data from 11 healthy adults, comparisons were made of EZP size estimations using oral and intravenous isotopes (r = 0.93 using urine) and urine and plasma enrichment (r = 0.85 for intravenous). EZP size estimations from urine and plasma enrichment following intravenous isotope administration were 2.35 +/- 0.84 and 2.83 +/- 0.86 mmol, respectively (mean +/- SD, P < 0.01). EZP size correlated with habitual dietary zinc intake (partial r = 0.74, P < 0.02). Cumulative declines in EZP size in two healthy adults after 3 wk of consuming a moderately zinc-restricted diet followed by 1 wk of severe zinc restriction were 26 and 32%. These results indicated that EZP size is dependent on dietary intake. We conclude that this technique is adequate to demonstrate EZP differences that are of nutritional and physiological interest. EZP size estimates can be obtained using orally or intravenously administered isotope and using plasma or urine enrichment data.

Intestinal fibrostenosis is among the hallmarks of severe Crohn’s disease. Patients with certain TNFSF15 (gene name for TL1A) variants over-express TL1A and have a higher risk of developing strictures in the small intestine. Additionally, sustained Tl1a expression in mice leads to small and large intestinal fibrostenosis under colitogenic conditions. The aim of this study was to determine whether established murine colonic fibrosis could be reversed with Tl1a antibody. Treatment with neutralizing Tl1a antibody reversed colonic fibrosis back to the original pre-inflamed levels, potentially as result of lowered expression of connective tissue growth factor (Ctgf), Il31Ra, transforming growth factor (Tgf) β1 and insulin-like growth factor-1 (Igf1). Additionally, blocking Tl1a function by either neutralizing Tl1a antibody or deletion of death domain receptor 3 (Dr3) reduced the number of fibroblasts and myofibroblasts, the primary cell types that mediate tissue fibrosis. Primary intestinal myofibroblasts expressed Dr3 and functionally responded to direct Tl1a signaling by increasing collagen and Il31Ra expression. These data demonstrated a direct role for TL1A-DR3 signaling in tissue fibrosis and that modulation of TL1A-DR3 signaling could inhibit gut fibrosis.

Summary HAP2, a male gamete-specific protein conserved across vast evolutionary distances has garnered considerable attention as a potential membrane fusogen required for fertilization in taxa ranging from protozoa and green algae to flowering plants and invertebrate animals [1–6]. However, its presence in Tetrahymena thermophilaa ciliated protozoan with seven sexes or mating types that bypasses the production of male gametes raises interesting questions regarding the evolutionary origins of gamete-specific functions in sexually dimorphic species. Here we show that HAP2 is expressed in all seven mating types of T. thermophila and that fertility is only blocked when the gene is deleted from both cells of a mating pair. HAP2 deletion strains of complementary mating types can recognize one another and form pairs, however pair stability is compromised and membrane pore formation at the nuclear exchange junction is blocked. The absence of pore formation is consistent with previous studies suggesting a role for HAP2 in gamete fusion in other systems. We propose a model in which each of the several hundred membrane pores established at the conjugation junction of mating Tetrahymena represents the equivalent of a male/female interface, and that pore formation is driven on both sides of the junction by the presence of HAP2. Such a model supports the idea that many of the disparate functions of sperm and egg were shared by the “isogametes” of early eukaryotes, and became partitioned to either male or female sex cells later in evolution.

Primary open angle glaucoma (POAG), a major cause of blindness worldwide, is a complex disease with a significant genetic contribution. We performed Exome Array (Illumina) analysis on 3504 POAG cases and 9746 controls with replication of the most significant findings in 9173 POAG cases and 26 780 controls across 18 collections of Asian, African and European descent. Apart from confirming strong evidence of association at CDKN2B-AS1 (rs2157719 [G], odds ratio [OR] = 0.71, P = 2.81 × 10−33), we observed one SNP showing significant association to POAG (CDC7–TGFBR3 rs1192415, ORG-allele = 1.13, Pmeta = 1.60 × 10−8). This particular SNP has previously been shown to be strongly associated with optic disc area and vertical cup-to-disc ratio, which are regarded as glaucoma-related quantitative traits. Our study now extends this by directly implicating it in POAG disease pathogenesis.

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