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Genome-wide association studies revealed that loss-of-function mutations in protein tyrosine phosphatase non-receptor type 2 (PTPN2) increase the risk of developing chronic immune diseases, such as inflammatory bowel disease (IBD) and celiac disease. These conditions are associated with increased intestinal permeability as an early etiological event. The aim of this study was to examine the consequences of deficient activity of the PTPN2 gene product, T cell protein tyrosine phosphatase (TCPTP), on intestinal barrier function and tight junction organization in vivo and in vitro. Here, we demonstrate that TCPTP protected against intestinal barrier dysfunction induced by the inflammatory cytokine IFN-γ by 2 mechanisms: it maintained localization of zonula occludens 1 and occludin at apical tight junctions and restricted both expression and insertion of the cation pore-forming transmembrane protein, claudin-2, at tight junctions through upregulation of the inhibitory cysteine protease, matriptase. We also confirmed that the loss-of-function PTPN2 rs1893217 SNP was associated with increased intestinal claudin-2 expression in patients with IBD. Moreover, elevated claudin-2 levels and paracellular electrolyte flux in TCPTP-deficient intestinal epithelial cells were normalized by recombinant matriptase. Our findings uncover distinct and critical roles for epithelial TCPTP in preserving intestinal barrier integrity, thereby proposing a mechanism by which PTPN2 mutations contribute to IBD.

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFr TKIs) are first-line therapies for various cancers, and cause dose-limiting severe diarrhea in many patients. We hypothesized that diarrhea caused by EGFr TKIs might reflect actions on epithelial transport, barrier function, or both, which we tested using cell cultures including murine and human enteroid-derived monolayers (EDMs), analyzed using electrophysiological and other relevant methods. EGFr TKIs (such as afatinib, erlotinib, and osimertinib) reversed the acute inhibitory effect of EGF on chloride secretion induced by carbachol (CCh) across T84 human colonic epithelial cells, which correlated with the diarrhea-inducing effect of each agent clinically. EGFr TKIs also reduced transepithelial electrical resistance (TEER), whereas co-treatment with CCh delayed the decrease in TEER compared with that of cells co-treated with EGF. Furthermore, afatinib and erlotinib prevented EGF- or CCh-induced EGFr phosphorylation. EGFr TKIs also suppressed phosphorylation of extracellular signal-regulated kinase (Erk)1/2 in response to EGF, whereas they had weaker effects on CCh-induced Erk1/2 phosphorylation. In human EDMs, EGF potentiated ion transport induced by CCh, whereas afatinib reversed this effect. The ability of EGFr TKIs to reverse the effects of EGF on calcium-dependent chloride secretion could contribute to the diarrheal side effects of these agents, and their disruption of epithelial barrier dysfunction is likely also pathophysiologically significant. CCh-activated Erk1/2 phosphorylation was relatively insensitive to EGFr TKIs and delayed the deleterious effects of EGFr TKIs on barrier function. These findings confirm and extend those of other authors, and may be relevant to designing strategies to overcome the diarrheal side effects of EGFr TKIs.

Congenital tufting enteropathy (CTE) is a rare chronic diarrheal disease of infancy caused by mutations in epithelial cell adhesion molecule (EpCAM). Previously, a murine CTE model showed mis-localization of EpCAM away from the basolateral cell surface in the intestine. Here we demonstrate that mutant EpCAM accumulated in the endoplasmic reticulum (ER) where it co-localized with ER chaperone, GRP78/BiP, revealing potential involvement of ER stress-induced unfolded protein response (UPR) pathway in CTE. To investigate the significance of ER-localized mutant EpCAM in CTE, activation of the three UPR signaling branches initiated by the ER transmembrane protein components IRE1, PERK, and ATF6 was tested. A significant reduction in BLOS1 and SCARA3 mRNA levels in EpCAM mutant intestinal cells demonstrated that regulated IRE1-dependent decay (RIDD) was activated. However, IRE1 dependent XBP1 mRNA splicing was not induced. Furthermore, an increase in nuclear-localized ATF6 in mutant intestinal tissues revealed activation of the ATF6-signaling arm. Finally, an increase in both the phosphorylated form of the translation initiation factor, eIF2α, and ATF4 expression in the mutant intestine provided support for activation of the PERK-mediated pathway. Our results are consistent with a significant role for UPR in gastrointestinal homeostasis and provide a working model for CTE pathophysiology.

Function diagrams put the focus on physiology and physiological concepts rather than the associated anatomy. Function diagrams could potentially serve as an elaboration tool and memory aid (mnemonic) to improve learning and recall. The function diagram prototype of the gastrointestinal system can aid in the instruction of difficult gastrointestinal physiology topics using a sequential focus on fundamental gastrointestinal functions.

Key points•Nucleotide binding oligomerization domain (Nod)-like receptors regulate cognition, anxiety and hypothalamic-pituitary-adrenal axis activation. •Nod-like receptors regulate central and peripheral serotonergic biology. •Nod-like receptors are important for maintenance of gastrointestinal physiology. •Intestinal epithelial cell expression of Nod1 receptors regulate behaviour.AbstractGut-brain axis signalling is critical for maintaining health and homeostasis. Stressful life events can impact gut-brain signalling, leading to altered mood, cognition and intestinal dysfunction. In the present study, we identified nucleotide binding oligomerization domain (Nod)-like receptors (NLR), Nod1 and Nod2, as novel regulators for gut-brain signalling. NLR are innate immune pattern recognition receptors expressed in the gut and brain, and are important in the regulation of gastrointestinal physiology. We found that mice deficient in both Nod1 and Nod2 (NodDKO) demonstrate signs of stress-induced anxiety, cognitive impairment and depression in the context of a hyperactive hypothalamic-pituitary-adrenal axis. These deficits were coupled with impairments in the serotonergic pathway in the brain, decreased hippocampal cell proliferation and immature neurons, as well as reduced neural activation. In addition, NodDKO mice had increased gastrointestinal permeability and altered serotonin signalling in the gut following exposure to acute stress. Administration of the selective serotonin reuptake inhibitor, fluoxetine, abrogated behavioural impairments and restored serotonin signalling. We also identified that intestinal epithelial cell-specific deletion of Nod1 (VilCre+ Nod1f/f ), but not Nod2, increased susceptibility to stress-induced anxiety-like behaviour and cognitive impairment following exposure to stress. Together, these data suggest that intestinal epithelial NLR are novel modulators of gut-brain communication and may serve as potential novel therapeutic targets for the treatment of gut-brain disorders.

BackgroundEosinophilic esophagitis (EoE) is a chronic TH2-assocated inflammatory condition accompanied by substantial impairments in epithelial barrier function and increased numbers of interleukin 9 (IL-9) expressing inflammatory cells. While IL-9 is known to affect barrier function in the intestine, the functional effects of IL-9 on the esophagus are unclear. Herein we aimed to understand the expression of the IL-9 receptor and effects of IL-9 on the epithelium in EoE.MethodsWe used esophageal biopsies from pediatric EoE patients with active and inactive disease to analyze the expression of the IL-9 receptor, the adherens junction protein E-cadherin and the tight junction protein claudin-1. We treated primary human esophageal epithelial cells with IL-9 to understand its effects on E-cadherin expression and function.ResultsActive EoE subjects had increased epithelial expression of IL-9 receptor mRNA and protein (P

Ontario, Canada mandates integrating environmental education (EE) into all subject areas from K-12, but pre-service teachers receive little to no instruction on how to do so. This study focusses on the experiences of 13 pre-service students who demonstrated their passion for EE through their own activism and volunteerism. Findings include the participants' definition of activism, lack of acknowledgement of activist experiences within the program, and participants' worries about being positioned as radical. Further, participants expressed a need for support, space, and time within the pre-service program to discuss these experiences and share ideas and resources within the teacher community. Suggestions to augment teacher education programs and for further research are provided.

In situ groundwater monitoring is critical for irrigated agroecosystems and informs land cover changes. Yet, such data can pose management challenges and confound agroecological relationships. Correspondingly, satellite-based approaches, including the GRACE-constellation, are increasing. Although in situ and GRACE-derived comparisons occur, limited research considers agroecological dependencies. Herein, we examined differences in groundwater monitoring approaches (observed [in situ, O] vs. predicted [GRACE-derived, P]) within the Yazoo–Mississippi Delta (YMD), an agroecosystem in the southeastern USA. We compared variations in modeled groundwater hydrology, land cover, and irrigation dynamics of the YMD within the upper-quartile (UQ) area of interest (AOI) (highest groundwater levels) and lower-quartile (LQ) AOI (lowest groundwater levels) every year from 2008 to 2020. Spatially, OUQ and PUQ were in northern portions of the YMD, with the OLQ and PLQ in southern portions. Groundwater levels between OUQ:PUQ and OLQ:PLQ each had correlations > 0.85. Regarding land cover, most categories varied within ±2.50% between model estimates over time. Relatedly, we documented 14 instances where correlations between land use category and groundwater level were inverted across models (OLQ:PLQ (5), OUQ:OLQ (6), PUQ:PLQ (3)). Irrigation results were not statistically different among all models. Overall, our results highlight the importance of quantifying model incongruences for groundwater and land cover management. [ABSTRACT FROM AUTHOR]

High school students' prevalence as food-service industry employees and their lack of food safety knowledge make them prime candidates for food safety education. The researchers developed a food-safety-focused curriculum for high school students aligned with Indiana Academic Standards for Agriculture, Advanced Life Science: Food. The curriculum was designed to provide students with fundamental food safety concepts through experiential learning and incorporation of science, technology, engineering, agriculture, and mathematics (STEAM) activities in the context of different careers related to agriculture, especially in food science. This study uses the Delphi technique to evaluate the food safety curriculum, including the identification of barriers to incorporating the curriculum into classrooms. The Delphi technique uses an expert panel to generate consensus related to a topic. A panel of experts in the field of education evaluated the curriculum through three rounds of surveys containing questions related to six curriculum assessment topics. Experts rated the degree to which they agreed with statements about the curriculum using a 5-point Likert scale and multiple-choice questions. At the conclusion of the study, the cost to purchase materials for cooking labs was the only identified barrier to curriculum incorporation (62.5%). Experts agreed that the curriculum addressed academic standards (100%), was engaging for students (100.0%), was easy for teachers to use (89.5%), and successfully incorporated STEAM (100.0%), experiential learning (89.5%), and career-education (78.9%). This study highlights the feasibility of providing food safety education to high school students in less traditional disciplines while promoting career development through the incorporation of experiential learning, STEAM, and career-education components.

This study explored children's area estimation performance. Two groups of fourth grade children completed area estimation tasks with rectangles ranging from 5 to 200 square units. A randomly assigned treatment group completed instructional sessions that involved a conceptual area measurement strategy along with numerical feedback. Children tended to underestimate areas of rectangles. Furthermore, rectangle size was related to performance such that estimation error and variability increased as rectangle size increased. The treatment group exhibited significantly improved area estimation performance in terms of accuracy, as well as reduced variability and instances of extreme responses. Area measurement estimation findings are related to a Hypothetical Learning Trajectory for area measurement.

While infectious agents have typical host preferences, the noninvasive enteric bacterium Vibrio cholerae is remarkable for its ability to survive in many environments, yet cause diarrheal disease (cholera) only in humans. One key V. cholerae virulence factor is its neuraminidase (VcN), which releases host intestinal epithelial sialic acids as a nutrition source and simultaneously remodels intestinal polysialylated gangliosides into monosialoganglioside GM1. GM1 is the optimal binding target for the B subunit of a second virulence factor, the AB5 cholera toxin (Ctx). This coordinated process delivers the CtxA subunit into host epithelia, triggering fluid loss via cAMP-mediated activation of anion secretion and inhibition of electroneutral NaCl absorption. We hypothesized that human-specific and human-universal evolutionary loss of the sialic acid N-glycolylneuraminic acid (Neu5Gc) and the consequent excess of N-acetylneuraminic acid (Neu5Ac) contributes to specificity at one or more steps in pathogenesis. Indeed, VcN was less efficient in releasing Neu5Gc than Neu5Ac. We show enhanced binding of Ctx to sections of small intestine and isolated polysialogangliosides from human-like Neu5Gc-deficient Cmah-/- mice compared to wild-type, suggesting that Neu5Gc impeded generation of the GM1 target. Human epithelial cells artificially expressing Neu5Gc were also less susceptible to Ctx binding and CtxA intoxication following VcN treatment. Finally, we found increased fluid secretion into loops of Cmah-/- mouse small intestine injected with Ctx, indicating an additional direct effect on ion transport. Thus, V. cholerae evolved into a human-specific pathogen partly by adapting to the human evolutionary loss of Neu5Gc, optimizing multiple steps in cholera pathogenesis.