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Background Although several studies link high levels of IL-6 and soluble IL-6 receptor (sIL-6R) to asthma severity and decreased lung function, the role of IL-6 trans-signaling (IL-6TS) in asthmatic patients is unclear. Objective We sought to explore the association between epithelial IL-6TS pathway activation and molecular and clinical phenotypes in asthmatic patients. Methods An IL-6TS gene signature obtained from air-liquid interface cultures of human bronchial epithelial cells stimulated with IL-6 and sIL-6R was used to stratify lung epithelial transcriptomic data (Unbiased Biomarkers in Prediction of Respiratory Disease Outcomes [U-BIOPRED] cohorts) by means of hierarchical clustering. IL-6TS–specific protein markers were used to stratify sputum biomarker data (Wessex cohort). Molecular phenotyping was based on transcriptional profiling of epithelial brushings, pathway analysis, and immunohistochemical analysis of bronchial biopsy specimens. Results Activation of IL-6TS in air-liquid interface cultures reduced epithelial integrity and induced a specific gene signature enriched in genes associated with airway remodeling. The IL-6TS signature identified a subset of patients with IL-6TS–high asthma with increased epithelial expression of IL-6TS–inducible genes in the absence of systemic inflammation. The IL-6TS–high subset had an overrepresentation of frequent exacerbators, blood eosinophilia, and submucosal infiltration of T cells and macrophages. In bronchial brushings Toll-like receptor pathway genes were upregulated, whereas expression of cell junction genes was reduced. Sputum sIL-6R and IL-6 levels correlated with sputum markers of remodeling and innate immune activation, in particular YKL-40, matrix metalloproteinase 3, macrophage inflammatory protein 1β, IL-8, and IL-1β. Conclusions Local lung epithelial IL-6TS activation in the absence of type 2 airway inflammation defines a novel subset of asthmatic patients and might drive airway inflammation and epithelial dysfunction in these patients. Graphical abstract [ABSTRACT FROM AUTHOR]

Methylisothiazolinone (MIT) is a common biocide used in cosmetic and industrial settings. Studies have demonstrated that MIT is a human sensitizer, to the extent that in 2013 MIT was named allergen of the year. Recently, we showed that MIT exposure in Xenopus laevis (the African clawed frog) inhibits wound healing and tail regeneration. However, it is unknown whether MIT affects these processes in other animals. Here, we investigated the effects of MIT exposure in planaria—non-parasitic freshwater flatworms able to regenerate all tissues after injury. Using a common research strain of Dugesia japonica , we determined that intact planarians exposed to 15 μM MIT displayed both neuromuscular and epithelial-integrity defects. Furthermore, regenerating (head and tail) fragments exposed to 15 μM MIT failed to close wounds or had significantly delayed wound healing. Planarian wounds normally close within 1 h after injury. However, most MIT-exposed animals retained open wounds at 24 h and subsequently died, and those few animals that were able to undergo delayed wound healing without dying exhibited abnormal regeneration. For instance, head regeneration was severely delayed or inhibited, with anterior structures such as eyes failing to form in newly produced tissues. These data suggest that MIT directly affects both wound healing and regeneration in planarians. Next, we investigated the ability of thiol-containing antioxidants to rescue planarian wound closure during MIT exposure. The data reveal both n -acetyl cysteine and glutathione were each able to fully rescue MIT inhibition of wound healing. Lastly, we established MIT toxicity levels by determining the LC 50 of 5 different planarian species: D. japonica, Schmidtea mediterranea , Girardia tigrina, Girardia dorotocephala, and Phagocata gracilis . Our LC 50 data revealed that concentrations as low as 39 μM (4.5 ppm) are lethal to planarians, with concentrations of just 5 μM inhibiting wound healing, and suggest that phylogeny is predictive of species toxicity levels. Together these results indicate MIT may have broad wound healing effects on aquatic species in general and are not limited to X. laevis alone. Future studies should investigate the impact of MIT on wound healing in other organisms, including non-aquatic organisms and mammals. [ABSTRACT FROM AUTHOR]

Alternative herbal therapy is increasingly used for inflammatory bowel disease (IBD), but it is essential to prove its safety and efficacy. We have assayed the anti-inflammatory effects of Amanda ® , a standardized water extract of Spanish Marcona almond skin very rich in polyphenols, in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. Amanda ® reduced the inflamed/ulcerated colonic area and preserved the mucosal architecture. The extract decreased the inflammatory infiltrate and the myeloperoxidase activity, as well as improved the oxidative state by increasing the glutathione content. Amanda ® also modulated the expression of pro-inflammatory cytokines and inducible nitric oxide synthase. Besides, Amanda ® improved the epithelial barrier function by restoring the levels of villin and mucin MUC3. Thus, the data from this work prove that Amanda ® extract possesses therapeutic activity in TNBS-induced colitis in rats, targeting different pathological factors of IBD like oxidative stress, neutrophil mucosal infiltration and loss of intestinal epithelial barrier function. [ABSTRACT FROM AUTHOR]

Summary: The palate is the ceiling of the oral cavity that separates the nasal and oral cavities in mammals. Anterior two-thirds of the palate that has the bone and the rugae on the oral surface is the hard palate, and the posterior one-third of the palate lacks bone and is referred to as the soft palate. Palate development involves a sequential process of outgrowth, elevation, and fusion of palatal shelves. When these steps are interrupted or compromised, cleft palate, one of the most frequent human congenital anomalies, develops. In particular, during the elevation process, sufficient proliferation of palatal mesenchymal cells and concomitant synthesis of extracellular matrix (ECM) are required, and these are referred to as intrinsic factors. One of major extrinsic factors for this process is the functional maturation of masticatory muscles that are responsible for movement of the mandible and lingual muscles, such that the tongue can descend and sufficient space for shelf elevation is provided. In addition to these factors, recent findings have suggested that a third factor, epithelial integrity, may be an essential component for successful shelf elevation. For example, disruption of epithelial integrity can result in ectopic epithelial fusion between the palate and the mandible, or tongue, which anchors the palatal shelf to inhibit its elevation. Although most histological and molecular aspects of palatal elevation have been well reviewed, those of epithelial integrity have not been thoroughly elucidated. Therefore, in this review, features of epithelial integrity, and its contributing molecules, are described in relation to palatal shelf elevation in mice. [Copyright &y& Elsevier]

Abstract: Antimicrobial peptides (AMPs) are a diverse group of proteinaceous compounds ranging in size, complexity and antimicrobial spectrum. The activity of AMPs against gut pathogens warrants the study of the interaction of AMPs with the mammalian gastrointestinal tract. In particular, the investigation of the in vitro cytotoxicity of these peptides is critical before they can be considered in clinical infections. The cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin was investigated in two gastrointestinal cell models (HT29 and Caco-2) with the MTT conversion assay, neutral red dye uptake assay and compared with that of vancomycin. The hemolytic activities were also investigated in sheep erythrocytes and the effect of AMPs on paracellular permeability was examined by transepithelial resistance (TEER) and TEM. Gallidermin was the least cytotoxic AMP followed by nisin A, magainin I, magainin II and melittin. Melittin and nisin were the only peptides to result in significant hemolysis. However, while nisin caused hemolysis at concentrations which were 1000-fold higher than those required for antimicrobial activity, melittin was hemolytic at concentrations in the same order of magnitude as its antimicrobial activity. Melittin was the only AMP to affect paracellular permeability. Long term melittin treatment also resulted in loss of microvilli, an increase in cell debris and destruction of intestinal tight junctions and cell–cell adhesion. Gallidermin shows most promise as a therapeutic agent, with relatively low cytotoxicity and potent antimicrobial activities. Melittin, while showing little potential as an antimicrobial agent, may have potential in delivery of poorly bioavailable drugs. [Copyright &y& Elsevier]

Evaluating the safety of previously fabricated and effective green synthetized colloidal silver (GSCS) on the mucosal barrier structure and function is essential prior to conduct human trials. The GSCS was applied to primary human nasal epithelial cells (HNECs) grown in an air-liquid interface (ALI) culture. Epithelial barrier integrity was evaluated by measuring the transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC)-dextran paracellular permeability. Ciliary beat frequency (CBF) was quantified. Effects of the GSCS on cell viability and inflammation were examined through lactate dehydrogenase, the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide viability assay and interleukin 6 (IL-6) enzyme linked immunosorbent assay. The localization and transportation of GSCS within HNECs and their HNEC-ALI cultures was assessed by transmission electron microscopy and inductively coupled plasma-mass-spectrometry, respectively. Application of GSCS to HNECs-ALI cultures for up to 2 h caused a significant reduction in the TEER values, however, it did not drop within the first 10 and 20 min for CRS and non-CRS control HNECs. The paracellular permeability, cell viability, IL-6 secretion and CBF remained unchanged. No GSCS was observed within or transported across HNECs. In conclusion, application of GSCS to HNECs is devoid of toxic effects. [ABSTRACT FROM AUTHOR]

Defects in the maintenance of intercellular junctions are associated with loss of epithelial barrier function and consequent pathological conditions, including invasive cancers. Epithelial integrity is dependent on actomyosin bundles at adherens junctions, but the origin of these junctional bundles is incompletely understood. Here we show that peripheral actomyosin bundles can be generated from a specific actin stress fiber subtype, transverse arcs, through their lateral fusion at cell-cell contacts. Importantly, we find that assembly and maintenance of peripheral actomyosin bundles are dependent on the mechanosensitive CaMKK2/AMPK signaling pathway and that inhibition of this route leads to disruption of tension-maintaining actomyosin bundles and re-growth of stress fiber precursors. This results in redistribution of cellular forces, defects in monolayer integrity, and loss of epithelial identity. These data provide evidence that the mechanosensitive CaMKK2/AMPK pathway is critical for the maintenance of peripheral actomyosin bundles and thus dictates cell-cell junctions through cellular force distribution. • Peripheral actomyosin bundles can be formed from a specific stress fiber subtype • Maintenance of these bundles is dependent on mechanosensitive CaMKK2/AMPK pathway • Inhibition of CaMKK2/AMPK pathway leads to redistribution of cellular forces Rajakylä et al. show that in epithelial cell cultures, peripheral actomyosin bundles are generated from a specific actin stress fiber type, transverse arcs. Bundle assembly is triggered by a mechanosensitive Ca2+/CaMKK2/AMPK pathway upon cell confluency. Inhibition of this route causes altered cellular force distribution and loss of epithelial cell identity. [ABSTRACT FROM AUTHOR]

Background & Aims It is a challenge to make a diagnosis of eosinophilic esophagitis (EoE) because its symptoms and histologic features overlap with those of gastroesophageal reflux disease (GERD). A minimally invasive device was recently developed to detect mucosal impedance (MI) that measures epithelial integrity during upper endoscopy. We aimed to quantify MI along the esophagus and identify patterns that differentiated patients with and without GERD from those with EoE, and determine whether MI values and patterns are sufficient to identify patients with EoE using histologic findings as a reference. Methods We performed a retrospective analysis of 91 patients with upper gastrointestinal symptoms referred for diagnostic testing for GERD and EoE from 2012 through 2014 (discovery set). During the first endoscopy, MI measurements were obtained at 2, 5, and 10 cm from the squamocolumnar junction. GERD was confirmed by ambulatory pH tests, and histologic analyses of biopsies were used to confirm EoE. We then used statistical modeling to identify MI patterns along the esophagus (at 10 cm, 5 cm, and 2 cm) that associated with GERD vs EoE. We validated our findings in a prospective cohort of 49 patients undergoing elective upper endoscopy for dysphagia, from 2015 through 2016, testing the ability of MI patterns to identify patients with vs. without EoE. Results We found patients with EoE to have a unique MI pattern, with low values along the esophageal axis. MI measurements at 5 cm could discern patients with normal vs abnormal mucosa with 83% sensitivity and 79% specificity, and patients with EoE vs GERD with 84% sensitivity and 70% specificity; these measurements differentiated the patient populations with the highest level of accuracy of any of the 6 measurements tested. In the validation study, a rater using the esophageal MI pattern identified patients with EoE with 100% sensitivity and 96% specificity. Conclusion We identified and validated a pattern of MI along the esophagus that can identify patients with EoE vs normal mucosa or GERD with high levels of sensitivity. [ABSTRACT FROM AUTHOR]