Gastrointestinal dysfunction in patients and mice expressing the autism-associated R451C mutation in neuroligin-3.

Gastrointestinal (GI) problems constitute an important comorbidity in many patients with autism. Multiple mutations in the neuroligin family of synaptic adhesion molecules are implicated in autism, however whether they are expressed and impact GI function via changes in the enteric nervous system is unknown. We report the GI symptoms of two brothers with autism and an R451C mutation in Nlgn3 encoding the synaptic adhesion protein, neuroligin-3. We confirm the presence of an array of synaptic genes in the murine GI tract and investigate the impact of impaired synaptic protein expression in mice carrying the human neuroligin-3 R451C missense mutation (NL3R451C ). Assessing in vivo gut dysfunction, we report faster small intestinal transit in NL3R451C compared to wild-type mice. Using an ex vivo colonic motility assay, we show increased sensitivity to GABAA receptor modulation in NL3R451C mice, a well-established Central Nervous System (CNS) feature associated with this mutation. We further show increased numbers of small intestine myenteric neurons in NL3R451C mice. Although we observed altered sensitivity to GABAA receptor modulators in the colon, there was no change in colonic neuronal numbers including the number of GABA-immunoreactive myenteric neurons. We further identified altered fecal microbial communities in NL3R451C mice. These results suggest that the R451C mutation affects small intestinal and colonic function and alter neuronal numbers in the small intestine as well as impact fecal microbes. Our findings identify a novel GI phenotype associated with the R451C mutation and highlight NL3R451C mice as a useful preclinical model of GI dysfunction in autism. Autism Res 2019, 12: 1043-1056. © 2019 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: People with autism commonly experience gastrointestinal problems, however the cause is unknown. We report gut symptoms in patients with the autism-associated R451C mutation encoding the neuroligin-3 protein. We show that many of the genes implicated in autism are expressed in mouse gut. The neuroligin-3 R451C mutation alters the enteric nervous system, causes gastrointestinal dysfunction, and disrupts gut microbe populations in mice. Gut dysfunction in autism could be due to mutations that affect neuronal communication.

TCF-1 limits the formation of Tc17 cells via repression of the MAF-RORγt axis.

Interleukin (IL)-17-producing CD8+ T (Tc17) cells have emerged as key players in host-microbiota interactions, infection, and cancer. The factors that drive their development, in contrast to interferon (IFN)-γ-producing effector CD8+ T cells, are not clear. Here we demonstrate that the transcription factor TCF-1 (Tcf7) regulates CD8+ T cell fate decisions in double-positive (DP) thymocytes through the sequential suppression of MAF and RORγt, in parallel with TCF-1-driven modulation of chromatin state. Ablation of TCF-1 resulted in enhanced Tc17 cell development and exposed a gene set signature to drive tissue repair and lipid metabolism, which was distinct from other CD8+ T cell subsets. IL-17-producing CD8+ T cells isolated from healthy humans were also distinct from CD8+IL-17- T cells and enriched in pathways driven by MAF and RORγt Overall, our study reveals how TCF-1 exerts central control of T cell differentiation in the thymus by normally repressing Tc17 differentiation and promoting an effector fate outcome.

Draft Genome Sequence of Enterobacter ludwigii NCR3, a Heavy Metal-Resistant Rhizobacterium.

We report here the draft genome of Enterobacter ludwigii NCR3, a Gram-negative bacterium isolated from the Carpobrotus rossii (Haw.) Schwantes rhizosphere. The analysis of the ~4.8-Mb draft genome shows that this strain harbors several genes associated with heavy metal resistance and plant growth-promoting activity, suggesting its potential application in microbe-assisted phytoremediation.

Responding to cough presentations: an interview study with Cambodian pharmacies participating in a National Tuberculosis Referral Program.

RATIONALE, AIMS AND OBJECTIVES: Asia-Pacific carries a high burden of respiratory-related mortality. Timely referral and detection of tuberculosis cases optimizes patient and public health outcomes. Registered private pharmacies in Cambodia participate in a National Tuberculosis Referral Program to refer clients with cough suggestive of tuberculosis to public sector clinics for diagnosis and care. The objective of this study was to investigate clinical intentions of pharmacy staff when presented with a hypothetical case of a client with prolonged cough suggestive of tuberculosis. METHOD: A random sample of 180 pharmacies was selected. Trained interviewers administered a hypothetical case scenario to trained pharmacy staff. Participants provided 'yes'/'no' responses to five clinical actions presented in the scenario. Actions were not mutually exclusive. Data were tabulated and compared using chi-square tests or Fisher's exact tests. RESULTS: Overall, 156 (92%) participants would have referred the symptomatic client in the case scenario. Participants who would have referred the client were less likely to sell a cough medicine (42% vs. 100%, P < 0.001) and less likely to sell an antibiotic (19% vs. 79%, P < 0.001) than those who would not have referred the client. CONCLUSION: Involving pharmacies in a Referral Program may have introduced concepts of appropriate clinical care when responding to clients presenting with cough suggestive of tuberculosis. However, results showed enhancing clinical competence among all referral programme participants particularly among non-referring pharmacies and those making concurrent sales of cough-related products would optimize pharmacy-initiated referral. Further research into actual clinical practices at Referral Program pharmacies would be justified.

Referral of tuberculosis symptomatic clients from private pharmacies to public sector clinics for diagnosis and treatment in Cambodia.

RATIONALE, AIMS AND OBJECTIVES: Cambodia is one of the 22 countries with a high burden of tuberculosis (TB). People often first seek treatment for cough and other TB symptoms through private pharmacies. The National Tuberculosis Programme trained willing private sector pharmacies to refer TB symptomatic clients to their closest public sector clinic for diagnosis and treatment. The study objective was to investigate factors associated with referral of TB symptomatic clients from pharmacies to public sector clinics in Phnom Penh, Cambodia. METHOD: Face-to-face structured interviews were conducted with staff from a stratified random sample of 180 private pharmacies in Phnom Penh in 2012. Trained interviewers were Khmer speakers. Logistic regression was used to compute odds ratios (ORs) and 95% confidence intervals (CIs) for factors associated with self-reported referral during the previous 3 months. RESULTS: Fifty (29.6%) pharmacies reported that they had referred 125 clients (range 1-10) to public sector clinics during the previous 3 months. In total, 164 (96.5%) pharmacies reported that they always referred all TB symptomatic clients to DOTS (directly observed treatment, short course) clinics. More than 6-year participation in the programme (OR 5.23, 95% CI 1.93-14.18) and willingness to always continue referring (OR 12.24, 95% CI 11.61-93.10) were associated with referral of one or more clients in the previous 3 months. Referral to the client's closest clinic was negatively associated with referral (OR 0.45, 95% CI 0.23-0.99). CONCLUSION: Pharmacies' ongoing commitment to the Referral Programme was strongly associated with referral. Increased advocacy among the high number of non-referring pharmacies may improve programme performance. Factors negatively associated with referral may need investigation.