There are an increasing variety of applications in which peptides are both synthesized and used attached to solid surfaces. This has created a need for high throughput sequence analysis directly on surfaces. However, common sequencing approaches that can be adapted to surface bound peptides lack the throughput often needed in library-based applications. Here we describe a simple approach for sequence analysis directly on solid surfaces that is both high speed and high throughput, utilizing equipment available in most protein analysis facilities. In this approach, surface bound peptides, selectively labeled at their N-termini with a positive charge-bearing group, are subjected to controlled degradation in ammonia gas, resulting in a set of fragments differing by a single amino acid that remain spatially confined on the surface they were bound to. These fragments can then be analyzed by MALDI mass spectrometry, and the peptide sequences read directly from the resulting spectra.
Recent infectious outbreaks highlight the need for platform technologies that can be quickly deployed to develop therapeutics needed to contain the outbreak. We present a simple concept for rapid development of new antimicrobials. The goal was to produce in as little as one week thousands of doses of an intervention for a new pathogen. We tested the feasibility of a system based on antimicrobial synbodies. The system involves creating an array of 100 peptides that have been selected for broad capability to bind and/or kill viruses and bacteria. The peptides are pre-screened for low cell toxicity prior to large scale synthesis. Any pathogen is then assayed on the chip to find peptides that bind or kill it. Peptides are combined in pairs as synbodies and further screened for activity and toxicity. The lead synbody can be quickly produced in large scale, with completion of the entire process in one week.
The labor, birth, and postpartum periods of women who experience stillbirth are physically similar to women with live birth; however, the negative effects are significantly greater.
The rise in antibiotic resistance has led to an increased research focus on discovery of new antibacterial candidates. While broad-spectrum antibiotics are widely pursued, there is evidence that resistance arises in part from the wide spread use of these antibiotics. Our group has developed a system to produce protein affinity agents, called synbodies, which have high affinity and specificity for their target. In this report, we describe the adaptation of this system to produce new antibacterial candidates towards a target bacterium. The system functions by screening target bacteria against an array of 10,000 random sequence peptides and, using a combination of membrane labeling and intracellular dyes, we identified peptides with target specific binding or killing functions. Binding and lytic peptides were identified in this manner and in vitro tests confirmed the activity of the lead peptides. A peptide with antibacterial activity was linked to a peptide specifically binding Staphylococcus aureus to create a synbody with increased antibacterial activity. Subsequent tests showed that this peptide could block S. aureus induced killing of HEK293 cells in a co-culture experiment. These results demonstrate the feasibility of using the synbody system to discover new antibacterial candidate agents.
Background: GoGirlGo! (GGG) is designed to increase girls’ physical activity (PA) using a health behavior and PA-based curriculum and is widely available for free to afterschool programs across the nation. However, GGG has not been formally evaluated. The purpose of this pilot study was to evaluate the effectiveness of the GGG curricula to improve PA, and self-efficacy for and enjoyment of PA in elementary aged girls (i.e., 5-13 years).
Methods: Nine afterschool programs were recruited to participate in the pilot (within subjects repeated measures design). GGG is a 12-week program, with a once a week, one-hour lesson with 30 minutes of education and 30 minutes of PA). Data collection occurred at baseline, mid (twice), post, and at follow-up (3-months after the intervention ended). PA was assessed via accelerometry at each time point. Self-efficacy for and enjoyment of PA was measured using the Self-Efficacy Scale and the Short-PA enjoyment scale and was assessed at baseline, post, and follow-up. Fidelity was assessed at midpoint.
Results: Across all age groups there was a statistically significant increase in PA. Overall, on days GGG was offered girls accumulated an average of 11 minutes of moderate-to-vigorous PA compared to 8 minutes during non-GGG days. There was a statistically significant difference in girls’ self-efficacy for PA reported between baseline and post, which was maintained at follow-up. An improvement in enjoyment of PA for girls was found between baseline and follow-up. According to fidelity assessment, 89% of the activities within the curriculum were completed each lesson. Girls appeared to respond well to the curriculum but girls 5-7 years had difficulties paying attention and understanding discussion questions.
Conclusions: Even though there were statistically significant differences in self-efficacy for PA and enjoyment of PA, minimal increases in girls’ PA were observed. GGG curricula improvements are warranted. Future GGG programming should explore offering GGG every day, modifying activities so that they are moderate-to-vigorous in intensity, and providing additional trainings that allow staff to better implement PA and improve behavior management techniques. With modifications, GGG could provide a promising no-cost curriculum that afterschool programs may implement to help girls achieve recommendations for PA.
Objective: This study aims to examine the relationship between friend advice about weight management, self-perception of weight, and alterations to weight change intentions, physical activity, and eating habits in college freshmen over time.
Methods: College freshmen from ASU with complete data for three time points (n=321) were found to be predominantly female (72.2%) and non-white (53.2%) with a mean age of 17.5±41. Complete data included responses for items included in analysis which were related to friend encouragement about weigh management, self-perception of weight, physical activity, eating behaviors, and weight change intentions. A longitudinal multivariate mediation analysis using negative binomial regression adjusted for sociodemographics and clustering by dorm was used to assess the relationship between 1) friend encouragement about weight management at time 1 and behavioral outcomes at time 3, 2) friend encouragement about weight management at time 1 and self-perception of weight at time 2, and 3) self-perception of weight at time 2 and behavioral outcomes at time 3.
Results: A small proportion of population perceived friend encouragement about weight loss (18.3%) and weight gain (14.4%) at time 1. Half the population (50.9%) had the self-perception of overweight at time 2. At time 3, more than half (54.3%) of individuals performed at least 60 minutes of MVPA and consumed at least ½ a serving of sugar-sweetened beverages each day, while nearly half (48.6%) consumed at least 2 servings of fruits and vegetables each day. Males perceived more friend encouragement to gain weight (27.4%; p<0.01), but more females had the self-perception of overweight (54%; p=0.04) and were attempting to lose weight (59.3%; p<0.01). Individuals who perceived friend encouragement to lose weight at time 1 had a 14.8% greater prevalence (p<0.001) of overweight perception of time two, and a 9.6% and 6.9%; decreased prevalence (p<0.001) of weight change and weight loss intentions (p=0.023) at time three respectively. Individuals who perceived friend encouragement to gain weight had a 34.9% decreased prevalence of (p<0.001) of self-perception of overweight at time 1. In individuals with the self-perception of overweight at time 2, there was a 18.1% increased prevalence (p<0.001) of consuming at least ½ a serving of sugar-sweetened beverages/day and an increased prevalence of 22.8% and 24.0% for weight change intentions and weight loss intentions at time 3 (p<0.001).
Conclusion: These findings suggest that there was not a mediation effect of self-perception of overweight in the relationship between friend encouragement about weight management and behavioral outcomes in the current sample. However, the increased prevalence of overweight perception in individuals who perceived friend encouragement about weight management may inform future interventions to focus on how weight-related conversations with friends is related to overweight perception. More research about the relationship between weight-related conversations had with friends, self-perception of weight, and health behaviors is needed to confirm these findings.
The heat-labile toxins (LT) produced by enterotoxigenic Escherichia coli display adjuvant effects to coadministered antigens, leading to enhanced production of serum antibodies. Despite extensive knowledge of the adjuvant properties of LT derivatives, including in vitro-generated non-toxic mutant forms, little is known about the capacity of these adjuvants to modulate the epitope specificity of antibodies directed against antigens. This study characterizes the role of LT and its non-toxic B subunit (LTB) in the modulation of antibody responses to a coadministered antigen, the dengue virus (DENV) envelope glycoprotein domain III (EDIII), which binds to surface receptors and mediates virus entry into host cells. In contrast to non-adjuvanted or alum-adjuvanted formulations, antibodies induced in mice immunized with LT or LTB showed enhanced virus-neutralization effects that were not ascribed to a subclass shift or antigen affinity. Nonetheless, immunosignature analyses revealed that purified LT-adjuvanted EDIII-specific antibodies display distinct epitope-binding patterns with regard to antibodies raised in mice immunized with EDIII or the alum-adjuvanted vaccine. Notably, the analyses led to the identification of a specific EDIII epitope located in the EF to FG loop, which is involved in the entry of DENV into eukaryotic cells. The present results demonstrate that LT and LTB modulate the epitope specificity of antibodies generated after immunization with coadministered antigens that, in the case of EDIII, was associated with the induction of neutralizing antibody responses. These results open perspectives for the more rational development of vaccines with enhanced protective effects against DENV infections.
Methods: This study was a randomized, wait-list, control trial with assessments at baseline and post-intervention (week 4). Participants were asked to meditate using Calm for 10 minutes per day. A p value ≤0.05 was considered statistically significant.
Results: The majority of participants (n=19) stated using Calm helped them cope with the stress of PA school. The intervention group participated in meditation for an average of 76 minutes/week. There were significant differences in all outcomes for the intervention group (all p ≤0.06). There was a significant interaction between group and time factors in emotional exhaustion (p=.016) and depersonalization (p=.025).
Conclusions: Calm is a feasible way to reduce burnout in PA students. Our findings provide information that can be applied to the design of future studies.