Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana.

Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The N-glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of N-glycans consists of complex type structures in both protein samples. The most abundant (>50%) N-glycan structure was GlcNAc₂(Xyl)Man₃(Fuc)GlcNAc₂ in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose N-glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in Nicotiana benthamiana plants with apoplast targeting

Comparing Programming Self-Esteem of Upper Secondary School Teachers to CS1 Students

Teacher self-esteem has been found to impact student learning in a number of non-computing fields. As computing slowly becomes a part of the upper secondary school (high school) curriculum in many countries, instruments designed to measure teachers’ programming self-esteem can help inform classroom practice and processes such as teacher professional development needs. This study examines if there are differences in programming self-esteem (using the Bergin Programming Self-Esteem Instrument) between upper secondary school teachers and CS1 students in Ireland. In addition this study provides evidence of validity when using this instrument (originally developed for CS1 students) to measure upper secondary school teacher programming self-esteem. To test for evidence of validity, we compared the results of the programming self-esteem construct given to upper secondary school teachers (n=130) to a recent study of programming selfesteem among CS1 students (n=693). We found evidence of both reliability and validity with teachers that aligns with the evidence found for the CS1 students, demonstrating utility for use with teacher cohorts. Comparing these findings, teachers reported statistically significantly lower programming self-esteem compared to CS1 students. Interestingly CS1 students identifying as male had a statistically significant higher programming self-esteem than those identifying as female. However, we found no statistically significant difference for teacher gender, unlike previous work. Our results indicate that teacher programming self-esteem should be given consideration in the design and implementation of professional development

Clinical lactation practice: 20 years of evidence

Lactation consultants depend on a vast multidisciplinary knowledge base to support their practices. To coincide with the 20-year anniversary of the International Lactation Consultant Association, the authors sought to highlight the knowledge base to demonstrate how practice has been affected. Using standard databases, they extracted English-language scientific literature related to breastfeeding and maternal and infant health outcomes; factors associated with breastfeeding initiation, exclusivity, and duration; lactation physiology; common breastfeeding challenges; breastfeeding practices within vulnerable populations; health professional support of breastfeeding; and breastfeeding practices in developing countries of Africa. Summaries of research are provided to demonstrate scientific method and knowledge evolution. As the knowledge of the biological, behavioral, and environmental factors that affect breastfeeding continues to grow, researchers and lactation consultants will identify additional research areas. Thus, the cycle of describing and explaining phenomena, testing interventions to improve practice, and ultimately improving breastfeeding outcomes worldwide will continue

Association Between Cytokines and Liver Histology in Children with Nonalcoholic Fatty Liver Disease.

BackgroundReliable non-invasive markers to characterize inflammation, hepatocellular ballooning, and fibrosis in nonalcoholic fatty liver disease (NAFLD) are lacking. We investigated the relationship between plasma cytokine levels and features of NAFLD histology to gain insight into cellular pathways driving NASH and to identify potential non-invasive discriminators of NAFLD severity and pattern.MethodsCytokines were measured from plasma obtained at enrollment in pediatric participants in NASH Clinical Research Network studies with liver biopsy-proven NAFLD. Cytokines were chosen a priori as possible discriminators of NASH and its components. Minimization of Akaike Information Criterion (AIC) was used to determine cytokines retained in multivariable models.ResultsOf 235 subjects, 31% had "Definite NASH" on liver histology, 43% had "Borderline NASH", and 25% had NAFLD but not NASH. Total plasminogen activator inhibitor 1 (PAI1) and activated PAI1 levels were higher in pediatric participants with Definite NASH and with lobular inflammation. Interleukin-8 (IL-8) was higher in those with stage 3-4 fibrosis and lobular inflammation. sIL-2rα was higher in children with stage 3-4 fibrosis and portal inflammation. In multivariable analysis, PAI1 variables were discriminators of Borderline/Definite NASH, definite NASH, lobular inflammation and ballooning. IL-8 increased with steatosis and fibrosis severity; sIL-2rα increased with fibrosis severity and portal inflammation. IL-7 decreased with portal inflammation and fibrosis severity.ConclusionsPlasma cytokines associated with histology varied considerably among NASH features, suggesting promising avenues for investigation. Future, more targeted analysis is needed to identify the role of these markers in NAFLD and to evaluate their potential as non-invasive discriminators of disease severity

Developing a realist informed framework for cultural adaptation of lifestyle interventions for the prevention of type 2 diabetes in South Asian populations in Europe

Aims Selected lifestyle interventions proven effective for White-European populations have been culturally adapted for South Asian populations living in Europe, who are at higher risk of type 2 diabetes. However, a limited theoretical basis underpins how cultural adaptations are believed to augment intervention effectiveness. We undertook a realist review to synthesise existing literature on culturally adapted type 2 diabetes prevention interventions, to develop a framework that shows ‘how’ cultural adaptation works, for ‘whom’ and in ‘what contexts’. Methods We followed the stepped methodological approach of realist review. Our work concluded a European-wide project (EuroDHYAN), and core studies were identified from the preceding EuroDHYAN reviews. Data were extracted, coded into themes and synthesised to create ‘Context–Mechanism–Outcome’ configurations and to generate a refined explanatory framework. Results We identified eight core intervention papers. From this evidence, and supporting literature, we examined the ‘Team’ domain of cultural adaptation and identified a mechanism of shared cultural identity which we theorised as contributing to strong team-participant relationships. We also identified four key contexts which influenced intervention outcomes: ‘research setting’ and ‘heterogeneous populations’ (intrinsic to the intervention) and ‘broader environment’ and ‘socio-cultural stress’ (extrinsic barriers). Conclusions This work instigates research into the mechanisms of cultural adaptation which, if pursued, will allow a more nuanced understanding of how to apply adaptations, and for whom. In practice we recommend greater consideration of heterogeneous and intersecting population characteristics; how intervention design can safeguard sustainability; and how the four key contexts identified influence how, and whether, these interventions work

Climate change and hydrology at the prairie margin : historic and prospective future flows of Canada's Red Deer and other Rocky Mountain river

This projected commenced with support from Alberta Agriculture and the Red Deer River Watershed Alliance, and subsequent funding was provided by Alberta Environment and Parks, Alberta Innovates, and the Natural Sciences and Engineering Research Council of Canada (NSERC), and Agriculture and Agri‐Food Canada provided some climate data. This paper follows from an MSc thesis chapter by the first author, and we extend thanks to faculty advisors Cam Goater, Stefan Kienzle, and Larry Flanagan and to two anonymous reviewers for very helpful recommendations.Peer reviewedPostprin

Evaluation of instream flow management for riparian restoration along the St. Mary River, Alberta

River damming and offstream water diversion have led to the severe collapse of riparian cottonwoods along the St. Mary River. To promote cottonwood recovery, a systemic restoration approach was applied by improving critical components of the flow regime, and initial assessments showed successful seedling recruitment. This research provides a longer-term assessment by comparing sapling distribution and growth among six regulated and free-flowing river reaches. Sapling patches along the lower St. Mary were very sparse (14% of reach) compared to those along the similarly-regulated Waterton River (82%). This disparity highlighted different conceptual models of degradation and restoration trajectories. Reach mean sapling diameters were strongly positively related to average spring river levels (R2 = 0.987), indicating that growth along the lower St. Mary may be hindered by low flows. However, more variation in sapling size occurred at smaller spatial scales indicating that site-level factors like density are also important determinants of growth

Assessment of abdominal fat compartments using DXA in premenopausal women from anorexia nervosa to morbid obesity

Objective: The purpose of this study was to test a newly developed DXA method for abdominal fat depot quantification in subjects with AN, normal weight, and obesity using CT as a gold standard. Design and Methods 135 premenopausal women (overweight/obese: n=89, normal-weight: n=27, AN: n=19); abdominal visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), and total adipose tissue (TAT) areas determined on CT and DXA. Results: There were strong correlations between DXA and CT measurements of abdominal fat compartments in all groups with the strongest correlation coefficients in the normal-weight and overweight/obese groups. Correlations of DXA and CT VAT measurements were strongest in the obese group and weakest in the AN group. DXA abdominal fat depots were higher in all groups compared to CT, with the largest % mean difference in the AN group and smallest in the obese group. Conclusions: A new DXA technique is able to assess abdominal fat compartments including VAT in premenopausal women across a large weight spectrum However, DXA measurements of abdominal fat were higher than CT, and this percent bias was most pronounced in the AN subjects and decreased with increasing weight, suggesting that this technique may be more useful in obese individuals

Optimization Of The Hockey Fans In Training (Hockey FIT) Weight Loss And Healthy Lifestyle Program For Male Hockey Fans

BACKGROUND: The health outcomes of men continue to be poorer than women globally. Challenges in addressing this problem include difficulties engaging men in weight loss programs as they tend to view these programs as contrary to the masculine narrative of independence and self-reliance. Researchers have been turning towards sports fans to engage men in health promotion programs as sports fans are typically male, and tend to have poor health habits. METHODS: Developed from the highly successful gender-sensitized Football Fans in Training program, Hockey Fans in Training (Hockey FIT) recruited 80 male hockey fans of the London Knights and Sarnia Sting who were overweight or obese into a weekly, 90-minute classroom education and group exercise program held over 12 weeks; a 40-week minimally-supported phase followed. A process evaluation of the Hockey FIT program was completed alongside a pragmatic randomized controlled trial and outcome evaluation in order to fully explore the acceptability of the Hockey FIT program from the perspectives of coaches delivering and participants engaged in the program. Data sources included attendance records, participant focus groups, coach interviews, assessment of fidelity (program observations and post-session coach reflections), and 12-month participant interviews. RESULTS: Coaches enjoyed delivering the program and found it simple to deliver. Men valued being among others of similar body shape and similar weight loss goals, and found the knowledge they gained through the program helped them to make and maintain health behaviour changes. Suggested improvements include having more hockey-related information and activities, greater flexibility with timing of program delivery, and greater promotion of technology support tools. CONCLUSIONS: We confirmed Hockey FIT was an acceptable gender-sensitized health promotion program for male hockey fans who were overweight or obese. Minor changes were required for optimization, which will be evaluated in a future definitive trial

The feasibility and effectiveness of a web-based personalised feedback and social norms alcohol intervention in UK university students: A randomised control trial

OBJECTIVE Alcohol misuse amongst University students is a serious concern, and research has started to investigate the feasibility of using e-health interventions. This study aimed to establish the effectiveness of an electronic web-based personalised feedback intervention through the use of a randomised control trial (RCT). METHODS 506 participants were stratified by gender, age group, year of study, self-reported weekly consumption of alcohol and randomly assigned to either a control or intervention condition. Intervention participants received electronic personalised feedback and social norms information on their drinking behaviour which they could access by logging onto the website at any time during the 12-week period. CAGE score, average number of alcoholic drinks consumed per drinking occasion, and alcohol consumption over the last week were collected from participants at pre- and post-survey. RESULTS A significant difference in pre- to post-survey mean difference of alcohol consumed per occasion was found, with those in the intervention condition displaying a larger mean decrease when compared to controls. No intervention effect was found for units of alcohol consumed per week or for CAGE scores. Sixty-three percent of intervention participants agreed that the feedback provided was useful. Those intervention participants who were above the CAGE cut off were more likely to report that the website would make them think more about the amount they drank. CONCLUSIONS Delivering an electronic personalised feedback intervention to students via the World Wide Web is a feasible and potentially effective method of reducing student alcohol intake. Further research is needed to replicate this outcome, evaluate maintenance of any changes, and investigate the process of interaction with web-based interventions