ICOS ligand and IL-10 synergize to promote host-microbiota mutualism.

Genome-wide association studies have identified ICOSLG, which encodes the inducible costimulator ligand (ICOSLG or ICOSL) as a susceptibility locus for inflammatory bowel disease. ICOSL has been implicated in the enhancement of pattern recognition receptor signaling in dendritic cells, induction of IL-10 production by CD4 T cells, and the generation of high-affinity antibodies to specific antigens-all of which can potentially explain its involvement in gastrointestinal inflammation. Here, we show that murine ICOSL deficiency results in significant enrichment of IL-10-producing CD4 T cells particularly in the proximal large intestine. Transient depletion of IL-10-producing cells from adult ICOSL-deficient mice induced severe colonic inflammation that was prevented when mice were first treated with metronidazole. ICOSL-deficient mice displayed reduced IgA and IgG antibodies in the colon mucus and impaired serum antibody recognition of microbial antigens, including flagellins derived from mucus-associated bacteria of the Lachnospiraceae family. Confirming the synergy between ICOSL and IL-10, ICOSL deficiency coupled with CD4-specific deletion of the Il10 gene resulted in juvenile onset colitis that was impeded when pups were fostered by ICOSL-sufficient dams. In this setting, we found that both maternally acquired and host-derived antibodies contribute to the life anti-commensal antibody repertoire that mediates this protection in early life. Collectively, our findings reveal a partnership between ICOSL-dependent anti-commensal antibodies and IL-10 in adaptive immune regulation of the microbiota in the large intestine. Furthermore, we identify ICOSL deficiency as an effective platform for exploring the functions of anti-commensal antibodies in host-microbiota mutualism.

Development and Evaluation of the Ally Sexual and Gender Minority Diversity and Inclusion Training at the Centers for Disease Control and Prevention.

CONTEXT: The Centers for Disease Control and Prevention (CDC) developed a workforce training on sexual and gender minorities (SGMs). OBJECTIVE: This article describes the evaluation of the training. DESIGN: Participants completed pre- and posttest surveys. After the pilot evaluation, some improvements were made to the curriculum and to the pre- and posttest questionnaires. Participants in subsequent (implementation) training were similarly asked to complete pre- and posttest questionnaires. SETTING: CDC. PARTICIPANTS: CDC staff. MAIN OUTCOME MEASURES: Participants' knowledge, ally identity, and perceptions of SGMs. RESULTS: Pilot and implementation training data showed increases in participant knowledge of 44% and 49%, respectively, increases in ally identity of 11% and 14%, respectively, and increases in positive perceptions of SGM of 25% and 31%, respectively. CONCLUSION: These results suggest that the CDC Ally Training may be a useful tool for improving staff knowledge and perceptions of SGM people.

DLG2 variants in patients with pubertal disorders.

PURPOSE: Impaired function of gonadotropin-releasing hormone (GnRH) neurons can cause a phenotypic spectrum ranging from delayed puberty to isolated hypogonadotropic hypogonadism (IHH). We sought to identify a new genetic etiology for these conditions. METHODS: Exome sequencing was performed in an extended family with autosomal dominant, markedly delayed puberty. The effects of the variant were studied in a GnRH neuronal cell line. Variants in the same gene were sought in a large cohort of individuals with IHH. RESULTS: We identified a rare missense variant (F900V) in DLG2 (which encodes PSD-93) that cosegregated with the delayed puberty. The variant decreased GnRH expression in vitro. PSD-93 is an anchoring protein of NMDA receptors, a type of glutamate receptor that has been implicated in the control of puberty in laboratory animals. The F900V variant impaired the interaction between PSD-93 and a known binding partner, Fyn, which phosphorylates NMDA receptors. Variants in DLG2 that also decreased GnRH expression were identified in three unrelated families with IHH. CONCLUSION: The findings indicate that variants in DLG2/PSD-93 cause autosomal dominant delayed puberty and may also contribute to IHH. The findings also suggest that the pathogenesis involves impaired NMDA receptor signaling and consequently decreased GnRH secretion.

Assessment of endoscope reprocessing at World Gastroenterology Organisation training centers using adenosine triphosphate testing.

Background and study aims Adequacy of endoscope disinfection in resource-limited settings is unknown. Adenosine triphosphate (ATP) testing is useful for evaluation of endoscope reprocessing, and ATP <200 relative light units (RLUs) after manual endoscope cleaning has been associated with adequacy of endoscope disinfection. Methods Consecutive endoscopes undergoing reprocessing at five World Gastroenterology Organisation (WGO) training centers underwent ATP testing before and after an on-site educational intervention designed to optimize reprocessing practices. Results A total of 343 reprocessing cycles of 65 endoscopes were studied. Mean endoscope age was 5.3 years (range 1-13 years). Educational interventions, based on direct observation of endoscope reprocessing practices at each site, included refinements in pre-cleaning, manual cleaning, high-level disinfection, and endoscope drying and storage. The percentage of reprocessing cycles with post-manual cleaning ATP ≧200 decreased from 21.4% prior to educational intervention to 14.8% post-intervention ( P =0.11). In multivariable logistic modelling, gastroscopes were significantly less likely (odds ratio [OR] 0.04, 95% confidence interval [CI] 0.01-0.19; P <0.001) than colonoscopes to achieve post-manual cleaning ATP < 200. No other factor (educational intervention, study site, endoscope age) was significantly associated with improved outcomes. Endoscope ID was not significantly associated with ATP values, and sites that performed manual versus automated HLD did not have significantly different likelihood of post-manual cleaning ATP <200 (OR 1.18, 95% CI 0.56-2.50; P =0.67). Conclusions In resource-limited settings, approximately 20% of endoscope reprocessing cycles may result in inadequate disinfection. This was not significantly improved by a comprehensive educational intervention. Alternative approaches to endoscope reprocessing are needed.

Early Life Stress in Mice Leads to Impaired Colonic Corticosterone Production and Prolonged Inflammation Following Induction of Colitis.

BACKGROUND: Early life stress (ELS) is an environmental trigger believed to promote increased risk of IBD. Our goal was to identify mechanisms whereby ELS in mice affects susceptibility to and/or severity of gut inflammation. METHODS: We utilized 2 published animal models of ELS. In the first model, newborn mice were separated from the dam daily for 4 to 8 hours starting on postnatal day 2 and then weaned early on postnatal day 17. Control mice were left undisturbed with the dams until weaning on postnatal day 21. In the second model, dams were fed dexamethasone or vehicle ad libitum in drinking water on postpartum days 1 to 14. Plasma and colonic corticosterone were measured in juvenile and adult mice. Colitis was induced in 4-week-old mice via intraperitoneal injection of interleukin (IL)-10 receptor blocking antibody every 5 days for 15 days. Five or 15 days later, colitis scores and transcripts for Tnf, glucocorticoid receptors, and steroidogenic enzymes were measured. RESULTS: Mice exposed to ELS displayed reduced plasma and colonic corticosterone. Control animals showed improvements in indices of inflammation following cessation of interleukin-10 receptor blockade, whereas ELS-exposed animals maintained high levels of Tnf and histological signs of colitis. In colitic animals, prior exposure to ELS was associated with significantly lower expression of genes associated with corticosterone synthesis and responsiveness. Finally, TNF stimulation of colonic crypt cells from ELS mice led to increased inhibition of corticosterone synthesis. CONCLUSIONS: Our study identifies impaired local glucocorticoid production and responsiveness as a potential mechanism whereby ELS predisposes to chronic colitis in susceptible hosts.

Using 2 distinct animal models, this study shows that in mice, early life stress leads to reduced colonic corticosterone and that induction of colitis after stress removal results in reduced transcription of glucocorticoid synthesis genes, increased Tnf, and enhanced chronicity of intestinal inflammation.

Haemobilia secondary to an arterio-biliary fistula: A rare complication of intra-operative microwave ablation of hepatocellular carcinoma.

A rare case of arterio-biliary fistula and haemobilia complicating intra-operative microwave ablation of hepatocellular carcinoma in a 58-year-old woman with cirrhosis.

mir-374-5p, mir-379-5p, and mir-503-5p Regulate Proliferation and Hypertrophic Differentiation of Growth Plate Chondrocytes in Male Rats.

Growth plate chondrocytes undergo sequential differentiation to form the resting zone, the proliferative zone (PZ), and the hypertrophic zone (HZ). The important role of microRNAs (miRNAs) in the growth plate was previously revealed by cartilage-specific ablation of Dicer, an enzyme essential for biogenesis of many miRNAs. To identify specific miRNAs that regulate differentiation of PZ chondrocytes to HZ chondrocytes, we microdissected individual growth plate zones from juvenile rats and performed miRNA profiling using a solution hybridization method and miRNA sequencing. Thirty-four miRNAs were differentially expressed between the PZ and the HZ, and we hypothesized that some of the miRNAs that are preferentially expressed in the PZ may promote proliferation and inhibit hypertrophic differentiation. Consistent with this hypothesis, transfection of inhibitors for four of these miRNAs (mir-369-3p, mir-374-5p, mir-379-5p, and mir-503-5p) decreased proliferation in primary epiphyseal chondrocytes. The inhibitors for three of these miRNAs (mir-374-5p, mir-379-5p, and mir-503-5p) also increased expression of multiple genes that are associated with chondrocyte hypertrophic differentiation. We next hypothesized that preferential expression of these miRNAs in the PZ is driven by the parathyroid hormone-related protein (PTHrP) concentration gradient across the growth plate. Consistent with this hypothesis, treatment of primary chondrocytes with a parathyroid hormone (PTH)/PTHrP receptor agonist, PTH1-34, increased expression of mir-374-5p, mir-379-5p, and mir-503-5p. Taken together, our findings suggest that the PTHrP concentration gradient across the growth plate induces differential expression of mir-374-5p, mir-379-5p, and mir-503-5p between the PZ and the HZ. In the PZ, the higher expression levels of these miRNAs promote proliferation and inhibit hypertrophic differentiation. In the HZ, downregulation of these miRNAs inhibits proliferation and promotes hypertrophic differentiation.

A case of an isolated gastric variceal bleed secondary to a pancreatic neuroendocrine tumour.

Gastric varices are an important complication of portal hypertension. We present here a case of bleeding gastric varices secondary to a pancreatic neuroendocrine tumour. To our knowledge, there are no other reports in the literature. Such cases are always a timely and important reminder of the importance in considering causes of portal hypertension other than cirrhosis, particularly in patients with limited risk factors for liver disease.

There is no dose-escalation response to dexamethasone (0.0625-1.0 mg/kg) in pediatric tonsillectomy or adenotonsillectomy patients for preventing vomiting, reducing pain, shortening time to first liquid intake, or the incidence of voice change.

BACKGROUND: Tonsillectomy is associated with postoperative nausea and vomiting (PONV) if no prophylaxis is administered. Previous studies have shown that a single dose of dexamethasone decreases the incidence of PONV. The most effective dose of dexamethasone to affect clinical outcome is yet to be defined. METHODS: One-hundred-twenty-five children were enrolled in a double-blind, prospective, randomized, dose-escalating study of dexamethasone: 0.0625, 0.125, 0.25, 0.5, or 1 mg/kg, maximum dose 24 mg. Nonparametric ANOVA was used to analyze the incidence of vomiting by treatment group for 0 to < or =5 h, >5 to 24 h. The Cox Proportional Likelihood Ratio Test was used to compare the time of first vomit and time to first pain medication across treatment groups. RESULTS: There was no difference in the incidence of vomiting for the five escalating doses of dexamethasone in the time period. There were no differences in secondary outcomes (analgesic requirements, time to first liquid, and change in voice) across treatment groups. CONCLUSION: We conclude that the lowest dose of dexamethasone (0.0625 mg/kg) was as effective as the highest dose of dexamethasone (1.0 mg/kg) for preventing PONV or reducing the incidence of other secondary outcomes following tonsillectomy or adenotonsillectomy. There is no justification for the use of high-dose dexamethasone for the prevention of PONV in this cohort of children.

Anatomical location of the bifurcation of the sciatic nerve in the posterior thigh in infants and children: a formula derived from MRI imaging for nerve localization.

BACKGROUND AND OBJECTIVES: Sciatic nerve block is performed at the popliteal fossa for various surgical procedures in infants and children. The aim of this study is to review magnetic resonance imaging scans in children of various ages to assess the location of the division of the nerve in the posterior thigh. METHODS: After Institutional Review Board approval was obtained, measurements of the bifurcation of the sciatic nerve in the posterior thigh were recorded from magnetic resonance images that were previously obtained in children of various ages. Data were recorded in a database and parametric and nonparametric statistical analysis was performed. RESULTS: Measurements were recorded from 59 patient images. Using a linear regression model, we were able to arrive at a formula for determining the point of bifurcation of the sciatic nerve at the posterior thigh. The mathematical formula derived from the linear regression equation was: CONCLUSIONS: We speculate that this knowledge will assist practitioners who use nerve stimulation techniques for sciatic nerve blocks in children of all ages.

Outcomes of preoperative biliary drainage from a single tertiary center: Is there still a role for plastic stents?

OBJECTIVES: Preoperative biliary drainage (PBD) can relieve symptoms of cholestasis, but carries risk of procedural complications. Metal stents have wider lumens and longer patency, although plastic stents (PS) remain in use. We reviewed the outcomes after PBD in patients with cholestasis. METHODS: Patients with symptomatic cholestasis who were likely to wait for over 2 weeks before surgery and were thus treated with PBD between January 2011 and May 2015 were included. Patients were evaluated for stenting-related complications, time interval to surgery, resection rate, improvement in bilirubin level and surgical complications. RESULTS: Forty patients underwent PBD by endoscopic retrograde cholangiopancreatography (ERCP). Of these, 36 patients received the placement of PS, one received a metal stent and the remaining three required percutaneous drainage due to unsuccessful biliary cannulation. Serum bilirubin declined from 172 µmol/L (baseline) to 14 µmol/L at 30 days (P < 0.0001). Median time interval from ERCP to surgery in all patients was 5 weeks (range 2-36 weeks). Preoperative stenting-related complications occurred in seven patients after a median of 3 weeks (range 1-6 weeks). Median time to surgery was similar in patients with and without stenting-related complications (5 weeks vs 4 weeks, respectively, P = 0.33). Surgery was completed in 32 (80%) patients, with a post-Whipple complication rate of 53%. CONCLUSIONS: PBD using mostly PS was effective in reducing bilirubin levels and did not detrimentally affect time interval to surgery. Median time interval to stenting-related complication occurred after 3 weeks, suggesting PS may be most useful for short-term PBD.

Endtidal carbon dioxide monitoring in children with congenital heart disease during sedation for cardiac catheterization by nonanesthesiologists.

BACKGROUND: This prospective observational study compared endtidal carbon dioxide (P(E)CO(2)) with blood gas carbon dioxide (PaCO(2)) values in children sedated by nonanesthesiologists for cardiac catheterization. METHODS: A nasal cannula designed to obtain gas sampling simultaneously from over the mouth and nares was taped into place after assuring a good waveform. Patients' cardiac lesions, site of blood gas sampling and P(E)CO(2)were recorded. RESULTS: Two hundred and one blood/P(E)CO(2) pairs were measured in 59 patients from 4 days to 18 years of age. Linear regression, Pearson correlation, and Bland-Altman analysis revealed a reasonable relationship (r = 0.493, P < 0.01, bivariate Pearson correlation) for all blood/expired CO(2) pairs even when the blood sample was obtained from an area of the circulation with shunting. There was no significant difference in the accuracy of the blood/P(E)CO(2) pairs between infants who weighed 15 kg. Thirteen children were diagnosed with partial or complete airway obstruction. CONCLUSIONS: Endtidal CO(2) measurement provides a reasonable reflection of blood CO(2) values if the expired gas-sampling catheter is taped in place after assuring a good waveform. The veracity of the data was the same throughout the patient size range. Expired CO(2) monitoring is useful for assessing the adequacy of respirations and the patency of the airway in children 3-89 kg.

"No! The lambs can stay out because they got cozies": constructive and destructive sibling conflict, pretend play, and social understanding.

Associations among constructive and destructive sibling conflict, pretend play, internal state language, and sibling relationship quality were investigated in 40 middle-class dyads with a kindergarten-age child (M age = 5.7 years). In 20 dyads the sibling was older (M age = 7.1 years) and in 20 dyads the sibling was younger (M age = 3.6 years). Dyads were videotaped playing with a farm set for 15 min; transcribed sessions were coded for (1) five types of conflict issues; (2) constructive, destructive, and passive resolution strategies; and (3) verbal and physical aggression. Measures of pretend play enactment, low- and high-level pretense negotiation strategies, and internal state language were also based on the transcripts. The Sibling Behavior and Feelings Questionnaire was used to assess both siblings' perceptions of sibling relationship quality. Findings revealed that conflict issues, aggression, and internal state language were associated with specific resolution strategies. Associations were evident between conflict issues and resolutions. Moreover, conflict issues and resolutions were associated with (1) relationship quality, (2) high-level pretense negotiation, and (3) internal state language employed in both play and conflict. Findings are discussed in light of recent theory on developmental processes operating within children's relationships.