Description: Activation of TLR4 by the endotoxin LPS is a critical event in the pathogenesis of Gram-negative sepsis. Caveolin-1, the signaling protein associated with caveolae, is implicated in regulating the lung inflammatory response to LPS; however, the mechanism is not understood. In this study, we investigated the role of caveolin-1 in regulating TLR4 signaling in endothelial cells. We observed that LPS interaction with CD14 in endothelial cells induced Src -dependent caveolin-1 phosphorylation at Tyr 14 . Using a TLR4-MD2-CD14–transfected HEK-293 cell line and caveolin-1–deficient ( cav-1 –/– ) mouse lung microvascular endothelial cells, we demonstrated that caveolin-1 phosphorylation at Tyr 14 following LPS exposure induced caveolin-1 and TLR4 interaction and, thereby, TLR4 activation of MyD88, leading to NF-B activation and generation of proinflammatory cytokines. Exogenous expression of phosphorylation-deficient Y14F caveolin-1 mutant in cav-1 –/– mouse pulmonary vasculature rendered the mice resistant to LPS compared with reintroduction of wild-type caveolin-1. Thus, caveolin-1 Y14 phosphorylation was required for the interaction with TLR4 and activation of TLR4-MyD88 signaling and sepsis-induced lung inflammation. Inhibiting caveolin-1 Tyr 14 phosphorylation and resultant inactivation of TLR4 signaling in pulmonary vascular endothelial cells represent a novel strategy for preventing sepsis-induced lung inflammation and injury.

Description: Objective There have been suggestions that currently recommended waist circumference (WC) cut-off points for Australians of European origin may not be applicable to Aboriginal people who have different body habitus profiles. We aimed to generate equivalent WC values that correspond to body mass index (BMI) points for identifying absolute cardiovascular disease (CVD) risks. Design Prospective cohort study. Setting An Aboriginal community in Australia's Northern Territory. Participants From 1992 to 1998, 920 adults without CVD, with age, WC and BMI measurements were followed-up for up to 20 years. Outcome measures Incident CVD, coronary artery disease (CAD) and heart failure (HF) events during the follow-up period ascertained from hospitalisation data. We generated WC values with 10-year absolute risks equivalent for the development of CVD as BMI values (20–34 kg/m 2 ) using the Weibull accelerated time-failure model. Results There were 211 incident cases of CVD over 13 669 person-years of follow-up. At the average age of 35 years, WC values with absolute CVD, CAD and HF risks equivalent to BMI of 25 kg/m 2 were 91.5, 91.8 and 91.7 cm, respectively, for males, and corresponding WC values were 92.5, 92.7 and 93 cm for females. WC values with equal absolute CVD, CAD and HF risks to BMI of 30 kg/m 2 were 101.7, 103.1 and 102.6 cm, respectively, for males, and corresponding values were 99.2, 101.6 and 101.5 cm for females. Association between WC and CVD did not depend on gender (p=0.54). Conclusions WC ranging from 91 to 93 cm was equivalent to BMI 25 kg/m 2 for overweight, and 99 to 103 cm was equivalent to BMI of 30 kg/m 2 for obesity in terms of predicting 10-year absolute CVD risk. Replicating the absolute risk method in other Aboriginal communities will further validate the WC values generated for future development of WC cut-off points for Aboriginal people.

Description: Objective To use Monte Carlo simulation to assess the uncertainty and variability of tobacco consumption through wastewater analysis in a city. Methods A total of 11 wastewater treatment plants (WWTPs) (serving 2.2 million people; approximately 83% of urban population in Dalian) were selected and sampled. By detection and quantification of principal metabolites of nicotine, cotinine (COT) and trans-3'-hydroxycotinine (OH-COT), in raw wastewater, back calculation of tobacco use in the population of WWTPs can be realised. Results COT and OH-COT were detected in the entire set of samples with an average concentration of 2.33±0.30 and 2.76±0.91 µg/L, respectively. The mass load of absorbed NIC during the sampling period ranged from 0.25 to 4.22 mg/day/capita with an average of 1.92 mg/day/capita. Using these data, we estimated that smokers in the sampling area consumed an average of 14.6 cigarettes per day for active smoker. Uncertainty and variability analysis by Monte Carlo simulation were used to refine this estimate: the procedure concluded that smokers in Dalian smoked between 10 and 27 cigarettes per day. This estimate showed good agreement with estimates from epidemiological research. Conclusions Sewage-based epidemiology may be a useful additional tool for the large-scale monitoring of patterns of tobacco use. Probabilistic methods can be used to strengthen the reliability of estimated use generated from wastewater analysis.

Description: Objectives We compared the differences in median age at spermarche among 11 ethnic minorities in 2010, estimated the trends regarding age at spermarche in different ethnic minorities from 1995 to 2010, and explored the association of spermarche with body mass index (BMI). Methods We used four cross-sectional Chinese National Surveys on Students’ Constitution and Health (CNSSCH, 1995, 2000, 2005 and 2010), and the total sample size was 40 113 children aged 11–18 years. The median age at spermarche of each ethnic minority was determined by using probit analysis. Logistic regression was used to assess the association of spermarche with BMI. Results In 2010, the ethnic minorities with earliest age at spermarche were Qiang (12.03 years), Zhuang (12.91 years) and Kirghiz (13.17 years); the three ethnic minorities with latest age at spermarche were Dong (14.73 years), Yao (14.60 years), and Naxi (14.36 years). From 1995 to 2010, age at spermarche showed a decline in almost each minority group except Yao and Dong. A higher BMI was associated with an increased likelihood of having reached spermarche after adjusting for age, regions or ethnic minorities. Conclusions A large variation in age at spermarche was observed among different ethnic minorities. The age at spermarche showed a downward shift in almost each of the 11 ethnic minorities with different patterns over time, and the children with higher BMI are more likely to enter puberty early.

Description: Objectives Regular exercise is a key component of obesity prevention and 48% of Americans do not meet minimum guidelines for weekly exercise. Social support has been shown to help individuals start and maintain exercise programmes. We evaluated social support among endurance athletes and explored the relationship between social support for exercise, health behaviours and health status. Design Survey. Setting The largest Nordic ski race in North America. Participants 5433 past participants responded to an online questionnaire. Outcome measures Social support, health behaviours and health status. Results The mean overall support score was 32.1 (SD=16.5; possible range=–16.0 to 88.0). The most common forms of social support were verbal such as discussing exercise, invitations to exercise and celebrating the enjoyment of exercise. We found that an increase of 10 points in the social support score was associated with a 5 min increase in weekly self-reported exercise (5.02, 95% CI 3.63 to 6.41). Conclusions Physical activity recommendations should incorporate the importance of participation in group activities, especially those connected to strong fitness cultures created by community and competitive events.

Description: Objective This study aimed to evaluate daily cholesterol intake across demographic factors and its food sources in elderly Chinese. Design A longitudinal study was conducted using demographic and dietary data for elders aged 60 and above from eight waves (1991–2011) of the China Health and Nutrition Survey. Setting The data were derived from urban and rural communities of nine provinces (autonomous regions) in China. Participants There were 16 274 participants (7657 male and 8617 female) in this study. Outcomes The primary outcome was daily cholesterol intake, which was calculated by using the Chinese Food Composition Table, based on dietary data. Results Daily consumption of cholesterol in the elderly significantly increased by 34% from 1991 to 2011 (p〈0.0001) and reached 253.9 mg on average in 2011. Secular trends in the proportion of subjects with an intake of 〉300 mg/day increased significantly during 1991–2011 (p〈0.0001). The major food sources of cholesterol by ranked order were eggs, pork, and fish and shellfish in 1991 and 2011, while organ meats which ranked fourth in the contribution to total intake in 1991 was replaced by poultry in 2011. Moreover, younger elders, male elders and elders from a high-income family or a highly urbanised community had higher cholesterol intakes and larger proportions of subjects with excessive cholesterol consumption in each survey year. Conclusions The large growth in daily cholesterol intake may pose major challenges for the health of elders in China. Reduced exposure to food enriched in cholesterol is required for elderly Chinese.

Description: Induction of type I IFN (IFN-I) is essential for host antiviral immune responses. However, IFN-I also plays divergent roles in antibacterial immunity, persistent viral infections, autoimmune diseases, and tumorigenesis. IFN regulatory factor 3 (IRF3) is the master transcription factor that controls IFN-I production via phosphorylation-dependent dimerization in most cell types in response to viral infections and various innate stimuli by pathogen-associated molecular patterns (PAMPs). To monitor the dynamic process of IRF3 activation, we developed a novel IRF3 dimerization reporter based on bimolecular luminescence complementation (BiLC) techniques, termed the IRF3-BiLC reporter. Robust induction of luciferase activity of the IRF3-BiLC reporter was observed upon viral infection and PAMP stimulation with a broad dynamic range. Knockout of TANK-binding kinase 1, the critical upstream kinase of IRF3, as well as the mutation of serine 386, the essential phosphorylation site of IRF3, completely abolished the luciferase activity of IRF3-BiLC reporter, confirming the authenticity of IRF3 activation. Taken together, these results demonstrated that the IRF3-BiLC reporter is a highly specific, reliable, and sensitive system to measure IRF3 activity. Using this reporter system, we further observed that the temporal pattern and magnitude of IRF3 activation induced by various PAMPs are highly complex with distinct cell type–specific characteristics, and IRF3 dimerization is a direct regulatory node for IFN-α/β receptor–mediated feed-forward regulation and crosstalk with other pathways. Therefore, the IRF3-BiLC reporter has multiple potential applications, including mechanistic studies as well as the identification of novel compounds that can modulate IRF3 activation.

Description: Helicobacter pylori infection causes chronic gastritis and peptic ulceration. H. pylori –initiated chronic gastritis is characterized by enhanced expression of many NF-B–regulated inflammatory cytokines. Brd4 has emerged as an important NF-B regulator and regulates the expression of many NF-B–dependent inflammatory genes. In this study, we demonstrated that Brd4 was not only actively involved in H. pylori –induced inflammatory gene mRNA transcription but also H. pylori –induced inflammatory gene enhancer RNA (eRNA) synthesis. Suppression of H. pylori –induced eRNA synthesis impaired H. pylori –induced mRNA synthesis. Furthermore, H. pylori stimulated NF-B–dependent recruitment of Brd4 to the promoters and enhancers of inflammatory genes to facilitate the RNA polymerase II–mediated eRNA and mRNA synthesis. Inhibition of Brd4 by JQ1 attenuated H. pylori –induced eRNA and mRNA synthesis for a subset of NF-B–dependent inflammatory genes. JQ1 also inhibited H. pylori –induced interaction between Brd4 and RelA and the recruitment of Brd4 and RNA polymerase II to the promoters and enhancers of inflammatory genes. Finally, we demonstrated that JQ1 suppressed inflammatory gene expression, inflammation, and cell proliferation in H. pylori –infected mice. These studies highlight the importance of Brd4 in H. pylori –induced inflammatory gene expression and suggest that Brd4 could be a potential therapeutic target for the treatment of H. pylori –triggered inflammatory diseases and cancer.

Description: Objective and design We conducted a systematic review and meta-analysis to evaluate the incidence of adverse events in the emergency department (ED) during procedural sedation in the paediatric population. Randomised controlled trials and observational studies from the past 10 years were included. We adhere to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Setting ED. Participants Children. Interventions Procedural sedation. Outcomes Adverse events like vomiting, agitation, hypoxia and apnoea. Meta-analysis was performed with random-effects model and reported as incidence rates with 95% CIs. Results A total of 1177 studies were retrieved for screening and 258 were selected for full-text review. 41 studies reporting on 13 883 procedural sedations in 13 876 children (≤18 years) were included. The most common adverse events (all reported per 1000 sedations) were: vomiting 55.5 (CI 45.2 to 65.8), agitation 17.9 (CI 12.2 to 23.7), hypoxia 14.8 (CI 10.2 to 19.3) and apnoea 7.1 (CI 3.2 to 11.0). The need to intervene with either bag valve mask, oral airway or positive pressure ventilation occurred in 5.0 per 1000 sedations (CI 2.3 to 7.6). The incidences of severe respiratory events were: 34 cases of laryngospasm among 8687 sedations (2.9 per 1000 sedations, CI 1.1 to 4.7; absolute rate 3.9 per 1000 sedations), 4 intubations among 9136 sedations and 0 cases of aspiration among 3326 sedations. 33 of the 34 cases of laryngospasm occurred in patients who received ketamine. Conclusions Serious adverse respiratory events are very rare in paediatric procedural sedation in the ED. Emesis and agitation are the most frequent adverse events. Hypoxia, a late indicator of respiratory depression, occurs in 1.5% of sedations. Laryngospasm, though rare, happens most frequently with ketamine. The results of this study provide quantitative risk estimates to facilitate shared decision-making, risk communication, informed consent and resource allocation in children undergoing procedural sedation in the ED.

Description: IgG autoantibodies mediate pathology in systemic lupus patients and lupus-prone mice. In this study, we showed that the class-switched IgG autoantibody response in MRL/ Fas lpr/lpr and C57/ Sle1Sle2Sle2 mice was blocked by the CID 1067700 compound, which specifically targeted Ras-related in brain 7 (Rab7), an endosome-localized small GTPase that was upregulated in activated human and mouse lupus B cells, leading to prevention of disease development and extension of lifespan. These were associated with decreased IgG-expressing B cells and plasma cells, but unchanged numbers and functions of myeloid cells and T cells. The Rab7 inhibitor suppressed T cell–dependent and T cell–independent Ab responses, but it did not affect T cell–mediated clearance of Chlamydia infection, consistent with a B cell–specific role of Rab7. Indeed, B cells and plasma cells were inherently sensitive to Rab7 gene knockout or Rab7 activity inhibition in class switching and survival, respectively, whereas proliferation/survival of B cells and generation of plasma cells were not affected. Impairment of NF-B activation upon Rab7 inhibition, together with the rescue of B cell class switching and plasma cell survival by enforced NF-B activation, indicated that Rab7 mediates these processes by promoting NF-B activation, likely through signal transduction on intracellular membrane structures. Thus, a single Rab7-inhibiting small molecule can target two stages of B cell differentiation to dampen the pathogenic autoantibody response in lupus.