An ABC transporter containing a forkhead-associated domain interacts with a serine-threonine protein kinase and is required for growth of Mycobacterium tuberculosis in mice

Forkhead-associated (FHA) domains are modular phosphopeptide recognition motifs with a striking preference for phosphothreonine-containing epitopes. FHA domains have been best characterized in eukaryotic signaling pathways but have been identified in six proteins in Mycobacterium tuberculosis, the causative organism of tuberculosis. One of these, coded by gene Rv1747, is an ABC transporter and the only one to contain two such modules. A deletion mutant of Rv1747 is attenuated in a mouse intravenous injection model of tuberculosis where the bacterial load of the mutant is 10-fold lower than that of the wild type in both lungs and spleen. In addition, growth of the mutant in mouse bone marrow-derived macrophages and dendritic cells is significantly impaired. In contrast, growth of this mutant in vitro was indistinguishable from that of the wild type. The mutant phenotype was lost when the mutation was complemented by the wild-type allele, confirming that it was due to mutation of Rv1747. Using yeast two-hybrid analysis, we have shown that the Rv1747 protein interacts with the serine-threonine protein kinase PknF. This interaction appears to be phospho-dependent since it is abrogated in a kinase-dead mutant and by mutations in the presumed activation loop of PknF and in the first FHA domain of Rv1747. These results demonstrate that the protein coded by Rv1747 is required for normal virulent infection by M. tuberculosis in mice and, since it interacts with a serine-threonine protein kinase in a kinase-dependent manner, indicate that it forms part of an important phospho-dependent signaling pathway

Crystallographic analysis of polypyrimidine tract-binding protein-Raver1 interactions involved in regulation of alternative splicing.

Accepted versio

Association between smoking cessation and short-term health-care use: results from an international prospective cohort study (ATTEMPT)

BACKGROUND AND AIMS: Previous studies have found that smoking cessation is associated with a short-term increase in health-care use. This may be because 'sicker' smokers are more likely to stop smoking. The current study assessed the association between smoking cessation and health-care use, adjusting for pre-cessation physical and mental health conditions. DESIGN/SETTING: Data came from the ATTEMPT cohort, a multi-national prospective survey of smokers in the United States, Canada, United Kingdom, France and Spain, that lasted 18 months (with follow-ups every 3 months). PARTICIPANTS: A total of 3645 smokers completed the baseline questionnaire. All participants smoked at least five cigarettes per day, intended to quit smoking within the next 3 months and were between 35 and 65 years of age. MEASUREMENTS: Participants were asked questions about their socio-demographic and smoking characteristics, as well previous smoking-related morbidities. Participants were also asked to report their health-care use in the previous 3 months i.e. emergency room (ER) visits, hospitalization, whether hospitalization required surgery, and health-care appointments. FINDINGS: A total of 8252, 4779 and 1954 baseline episodes of smoking were available for 3, 6 and 12 months, respectively. Of these, 2.8% (n = 230), 0.9% (n = 40) and 0.7% (n = 14) were followed by 3, 6 and 12 months of abstinence. No significant differences were found among 3, 6 or 12 months of abstinence and ER visits, hospitalization and whether hospitalization required surgery or health-care visits. However, 6-month smoking cessation episodes were associated with higher odds of reporting an appointment with a dietician. CONCLUSION: Smoking cessation does not appear to be associated with a substantial short-term increase or decrease in health-care use after adjusting for pre-cessation morbidities

Primary versus Staged Closure of Exomphalos Major: Cardiac Anomalies Do Not Affect Outcome

Aim: The objective of the study is to describe management of exomphalos major and investigate the effect of congenital cardiac anomalies. / Methods: A single-center retrospective review (with audit approval) was performed of neonates with exomphalos major (fascial defect ≥ 5cm ± liver herniation) between 2004 and 2014. Demographic and operative data were collected and outcomes compared between infants who had primary or staged closure. Data, median (range), were analyzed appropriately. Results: A total of 22 patients were included, 20 with liver herniation and 1 with pentalogy of Cantrell. Gestational age was 38 (30–40) weeks, birth weight 2.7 (1.4–4.6) kg, and 13 (60%) were male. Two were managed conservatively due to severe comorbidities, 5 underwent primary closure, and 15 had application of Prolene (Ethicon Inc) mesh silo and serial reduction. Five died, including two managed conservatively, none primarily of the exomphalos. Survivors were followed up for 38 months (2–71). Cardiac anomalies were present in 20 (91%) patients: 8 had minor and 12 major anomalies. Twelve (55%) patients had other anomalies. Primary closure was associated with shorter length of stay (13 vs. 85 days, p = 0.02), but infants had similar lengths of intensive care stay, duration of parenteral feeds, and time to full feeds. Infants with cardiac anomalies had shorter times to full closure (28 vs. 62 days, p = 0.03), but other outcomes were similar. / Conclusion: Infants whose defect can be closed primarily have a shorter length of stay, but other outcomes are similar. Infants with more significant abdominovisceral disproportion are managed with staged closure; the presence of major cardiac anomalies does not affect surgical outcome

Comparison of clinic-based versus home-based balance and agility training for the symptoms of knee osteo-arthritis

Objective. To compare clinic-based (CB) and home-based (HB) deliveries of a knee osteoarthritis (OA) exercise programme. Methods. Outcomes from a CB exercise study (N=6) utilising kinesthesia, balance and agility (KBA) exercises were compared with those from a HB KBA study (N=6). Both conditions trained 30 minutes, 3 days per week for 8 weeks. CB sessions were conducted in a group led by an exercise physiologist (EP); HB participants received an initial 3 sessions of one-to-one training from an EP, written/pictorial instructions, telephone and e-mail follow-up, and in-person refresher sessions during weeks 4 and 6. The primary outcome was an OA-specific physical function survey. Community activity level, self-report knee stability, 15-m get up and go walk, and stair climb and descent were also measured. Results. Adherence was 94% in both conditions. KBA improved PF in both CB (59%; 18±12.5 pts; p=0.008) and HB (33%; 7.3±7.5 pts; p=0.03), with no difference between conditions. All outcome improvements were somewhat larger for CB, but these differences did not reach statistical significance. Conclusion. We found no difference in outcomes between CB and HB exercise in this preliminary comparison. Our results support that KBA is an effective intervention for symptomatic knee OA that may be delivered in CB or HB settings

Policy Coherence in US Tobacco Control: Beyond FDA Regulation

Joshua Yang and Thomas Novotny explore whether the US government can develop and implement a coherent policy agenda to reduce tobacco-related morbidity and mortality

Predictors of Segmented School Day Physical Activity and Sedentary Time in Children from A Northwest England Low-income Community

Background: Schools have been identified as important settings for health promotion through physical activity participation, particularly as children are insufficiently active for health. The aim of this study was to investigate the child and school-level influences on children′s physical activity levels and sedentary time during school hours in a sample of children from a low-income community; Methods: One hundred and eighty-six children (110 boys) aged 9–10 years wore accelerometers for 7 days, with 169 meeting the inclusion criteria of 16 h∙day−1 for a minimum of three week days. Multilevel prediction models were constructed to identify significant predictors of sedentary time, light, and moderate to vigorous physical activity during school hour segments. Child-level predictors(sex, weight status, maturity offset, cardiorespiratory fitness, physical activity self-efficacy, physical activity enjoyment) and school-level predictors (number on roll, playground area, provision score) were entered into the models; Results: Maturity offset, fitness, weight status, waist circumference-to-height ratio, sedentary time, moderate to vigorous physical activity, number of children on roll and playground area significantly predicted physical activity and sedentary time; Conclusions: Research should move towards considering context-specific physical activity and its correlates to better inform intervention strategies

Evaluation of wrist and hip sedentary behaviour and moderate-to-vigorous physical activity raw acceleration cutpoints in older adults

Wrist-based accelerometers are now common in assessing physical activity (PA) and sedentary behaviour (SB) in population-based studies, but there is a scarcity of raw acceleration cutpoints in older adults. The study aimed to determine and evaluate wrist-based GENEActiv (GA) and hip-based ActiGraph GT3X+ (AG) raw acceleration cutpoints for SB and moderate-to-vigorous PA (MVPA) in older adults ≥60 years of age. A laboratory-based calibration analyses of 34 healthy older adults involved receiver operator characteristic (ROC) curves to determine raw acceleration cutpoints for SB and MVPA. ROC analysis revealed an area under the curve (AUC) of 0.88 for GA SB and MVPA, and 0.90 for AG SB and 0.94 for AG MVPA. Sensitivity optimised SB and specificity optimised MVPA GA cutpoints of 57 mg and 104 mg, and AG cutpoints of 15 mg and 69 mg were also generated, respectively. Cross-validation analysis revealed moderate agreement for GA and AG SB cutpoints, and fair to substantial agreement for GA and AG MVPA cutpoints, respectively. The resultant cutpoints can classify older adults as engaging in SB or not engaging in MVPA but the sensitivity optimised SB cutpoints should be interpreted with a degree of caution due to their modest cross-validation results

Surgical jejunostomy and radiological gastro-jejunostomy feeding in children: Risks, benefits and nutritional outcomes

PURPOSE: Radiologically inserted gastrojejunal tubes (RGJ) and surgical jejunostomy (SJ) are established modes of jejunal feeding. The aim of the study is to review nutritional outcomes, complications and the practical consideration to enable patients and carers to make informed choice. METHODS: Retrospective review of patient notes with a RGJ or SJ in 2010, with detailed follow-up and review of the literature. RESULTS: Both RGJ and SJ are reliable modes to provide stable enteral nutrition. Both have complications and their own associated limitations. CONCLUSION: The choice has to be tailored to the individual patient, the social care available, the inherent medical disease and risk/benefit of repeated anaesthetic and radiation exposure. RGJ and SJ are important tools for nutritional management that achieve and maintain growth in a complex group of children. The risk and benefits should be reviewed for each individual patient

Reproducible isolation of lymph node stromal cells reveals site-dependent differences in fibroblastic reticular cells

Within lymph nodes, non-hematopoietic stromal cells organize and interact with leukocytes in an immunologically important manner. In addition to organizing T and B cell segregation and expressing lymphocyte survival factors, several recent studies have shown that lymph node stromal cells shape the naïve T cell repertoire, expressing self-antigens which delete self-reactive T cells in a unique and non-redundant fashion. A fundamental role in peripheral tolerance, in addition to an otherwise extensive functional portfolio, necessitates closer study of lymph node stromal cell subsets using modern immunological techniques; however this has not routinely been possible in the field, due to difficulties reproducibly isolating these rare subsets. Techniques were therefore developed for successful ex vivo and in vitro manipulation and characterization of lymph node stroma. Here we discuss and validate these techniques in mice and humans, and apply them to address several unanswered questions regarding lymph node composition. We explored the steady-state stromal composition of lymph nodes isolated from mice and humans, and found that marginal reticular cells and lymphatic endothelial cells required lymphocytes for their normal maturation in mice. We also report alterations in the proportion and number of fibroblastic reticular cells (FRCs) between skin-draining and mesenteric lymph nodes. Similarly, transcriptional profiling of FRCs revealed changes in cytokine production from these sites. Together, these methods permit highly reproducible stromal cell isolation, sorting, and culture