Epitope mapping using mRNA display and a unidirectional nested deletion library

In vitro selection targeting an anti-polyhistidine monoclonal antibody was performed using mRNA display with a random, unconstrained 27-mer peptide library. After six rounds of selection, epitope-like peptides were identified that contain two to five consecutive, internal histidines and are biased for arginine residues, without any other identifiable consensus. The epitope was further refined by constructing a high-complexity, unidirectional fragment library from the final selection pool. Selection by mRNA display minimized the dominant peptide from the original selection to a 15-residue functional sequence (peptide Cmin: RHDAGDHHHHHGVRQ; K-D = 38 nM). Other peptides recovered from the fragment library selection revealed a separate consensus motif (ARRXA) C-terminal to the histidine track. Kinetics measurements made by surface plasmon resonance, using purified Fab (antigen-binding fragment) to prevent avidity effects, demonstrate that the selected peptides bind with 10- to 75-fold higher affinities than a hexahistidine peptide. The highest affinity peptides (K-D approximate to 10 nM) encode both a short histidine track and the ARRXA motif, suggesting that the motif and other flanking residues make important contacts adjacent to the core polyhistidine-binding site and can contribute > 2.5 kcal/mol of binding free energy. The fragment library construction methodology described here is applicable to the development of high-complexity protein or cDNA expression libraries for the identification of protein-protein interaction domains

Delivering manufacturing technology and workshop appreciation to engineering undergraduates using the flipped classroom approach

Delivery of manufacturing technology and practical workshop-based work, on undergraduate engineering courses that engage the learners, is challenging. The paper presents an experimental method of workshop delivery using the flipped learning approach, a pedagogical model in which the typical lecture and homework elements of a course are reversed. Video lectures are viewed by students prior to class. In-class time can be devoted to exercises, projects, or discussions as in this case. Learners were asked to observe three audiovisual clips in preparation for class. The objective was to determine whether the flipped classroom approach can enhance the learning experience, through better engagement with the students, compared to conventional classroom-based learning. The level of student participation and level of success have been established by means of feedback questionnaires from more than 100 participants and peer observation. The results are encouraging and demonstrate that this approach is favoured by the students

A Peptide Core Motif for Binding to Heterotrimeric G Protein α Subunits

Recently, in vitro selection using mRNA display was used to identify a novel peptide sequence that binds with high affinity to G{alpha}i1. The peptide was minimized to a 9-residue sequence (R6A-1) that retains high affinity and specificity for the GDP-bound state of G{alpha}i1 and acts as a guanine nucleotide dissociation inhibitor (GDI). Here we demonstrate that the R6A-1 peptide interacts with G{alpha} subunits representing all four G protein classes, acting as a core motif for G{alpha} interaction. This contrasts with the consensus G protein regulatory(GPR) sequence, a 28-mer peptide GDI derived from the GoLoco (G{alpha}i/0-Loco interaction)/GPR motif that shares no homology with R6A-1 and binds only to G{alpha}i1-3 in this assay. Binding of R6A-1 is generally specific to the GDP-bound state of the G{alpha} subunits and excludes association with G{beta}{gamma}. R6A-G{alpha}i1 complexes are resistant to trypsin digestion and exhibit distinct stability in the presence of Mg2+, suggesting that the R6A and GPR peptides exert their activities using different mechanisms. Studies using G{alpha}i1/G{alpha}s chimeras identify two regions of G{alpha}i1 (residues 1–35 and 57–88) as determinants for strong R6A-Gi{alpha}1 interaction. Residues flanking the R6A-1 peptide confer unique binding properties, indicating that the core motif could be used as a starting point for the development of peptides exhibiting novel activities and/or specificity for particular G protein subclasses or nucleotide-bound states

A CORRELATIONAL-COMPARATIVE STUDY OF GRADE 4, GRADE 5, AND GRADE 6 STUDENTS’ SELF-EFFICACY FOR LEARNING AND PERFORMANCE IN INTEGRATED SUBJECTS WITH THEIR ACADEMIC ACHIEVEMENT IN A MOTHER TONGUE-BASED MULTILINGUAL EDUCATION CONTEXT IN 5 SJN-CBE SCHOOLS IN

The purpose of this quantitative study was to investigate whether there was a significant relationship between self-efficacy for learning and performance in integrated subjects of Grade 4, Grade 5, and Grade 6 students and their academic achievement in 5 SJN-CBE Schools, Northern State, Myanmar. An adapted version of the Motivated Strategies for Learning Questionnaire by Pintrich, Smith, Gracia, and McKeachie (1991) was used to collect data from a total of 433 students; 117 Grade 4 students, 159 Grade 5 students, and 157 Grade 6 students. The subjects' final assessment results were used to determine the integrated subjects' academic achievement levels. The researcher also compared the self-efficacy for learning and performance in integrated subjects among Grade 4, Grade 5, and Grade 6 students. Descriptive statistics (means and standard deviations) and statistical hypothesis testing (correlational analysis using Pearson product-moment correlation and one-way ANOVA) were used to analyze the data. The research findings indicated high levels of self-efficacy for learning and performance in integrated subjects of Grade 4, Grade 5, and Grade 6. The integrated subject's final assessment result revealed that grade 4, Grade 5, and Grade 6 students had good academic achievement. Pearson product-moment correlation suggested that self-efficacy for learning and performance in integrated subjects of Grade 5 and grade 6 students and their academic achievement were significantly positively correlated; in contrast, there was no significant relationship between students' self-efficacy for learning and performance in integrated subjects of Grade 4 students and their academic achievement. The findings indicated a significant1 M.Ed. in Curriculum & Instruction, Principal, Church-Based Education Shan State Kachin Baptist Union, Loihkang Road, Kawnghka, Kutkai, Northern Shan State, Myanmar. [email protected] Ph.D., Assistant Professor, Graduate School of Human Sciences, Assumption University, Thailand. [email protected]: Human Sciences, ISSN 2586-9388, Vol.14 No.2 (Jul.-Dec. 2022)difference in self-efficacy for learning and performance in integrated subjects between Grade 4 and Grade 5. Still, there was no significant difference between Grade 4 and Grade 6 students and between Grade 5 and Grade 6 students. Recommendations for students, teachers, parents, implementers and administrators of SJN-CBE, and future researchers are provided

Learning to laugh : children and being human in early modern thought

This essay explores the construction of the human in early modern English thought, and uses discussions of the nature and use of laughter as a distinguishing feature of humanity from classical arguments as well as early modern ones. Using these classical, reformed English discussions of education and of the nature of children reveals an anxiety about the status of the child. Laughing appropriately - using tile mind and not merely the body - is a key feature of being human, and as such, the child's lack of "true' laughter reveals that child's status to be never always-already human. "Human' is a created rather than merely a natural status

Comparative visual function in five sciaenid fishes inhabiting Chesapeake Bay

Maintaining optimal visual performance is a difficult task in the photodynamic coastal and estuarine waters in which western North Atlantic sciaenid fishes support substantial commercial and recreational fisheries. Unavoidable tradeoffs exist between visual sensitivity and resolution, yet sciaenid visual systems have not been characterized despite strong species-specific ecomorphological and microhabitat differentiation. We therefore used electroretinographic techniques to describe the light sensitivities, temporal properties, and spectral characteristics of the visual systems of five sciaenids common to Chesapeake Bay, USA: weakfish (Cynoscion regalis), spotted seatrout (Cynoscion nebulosus), red drum (Sciaenops ocellatus), Atlantic croaker (Micropogonias undulatus) and spot (Leiostomus xanthurus). Benthic sciaenids exhibited higher sensitivities and broader dynamic ranges in white light V/logI experiments than more pelagic forms. Sensitivities of the former were at the lower (more sensitive) end of an emerging continuum for coastal fishes. Flicker fusion frequency experiments revealed significant interspecific differences at maximum intensities that correlated with lifestyle and habitat, but no specific differences at dimmer intensities. Spectral responses of most sciaenids spanned 400-610 nm, with significant diel differences in weakfish and Atlantic croaker. Weakfish, a crepuscular predator, also responded to ultraviolet wavelengths; this characteristic may be more useful under less turbid conditions. Collectively, these results suggest that sciaenids are well adapted to the dynamic photoclimate of the coastal and estuarine waters they inhabit. However, the recent anthropogenic degradation of water quality in coastal environments, at a pace faster than the evolution of visual systems, has amplified the importance of characterizing visual function in managed aquatic fauna

Acoustic pressure and particle motion thresholds in six sciaenid fishes

Sciaenid fishes are important models of fish sound production, but investigations into their auditory abilities are limited to acoustic pressure measurements on five species. In this study, we used auditory brainstem response ( ABR) to assess the pressure and particle acceleration thresholds of six sciaenid fishes commonly found in Chesapeake Bay, eastern USA: weakfish ( Cynoscion regalis), spotted seatrout ( Cynoscion nebulosus), Atlantic croaker ( Micropogonias undulatus), red drum ( Sciaenops ocellatus), spot ( Leiostomus xanthurus) and northern kingfish ( Menticirrhus saxatilis). Experimental subjects were presented with pure 10 ms tone bursts in 100 Hz steps from 100 Hz to 1.2 kHz using an airborne speaker. Sound stimuli, monitored with a hydrophone and geophone, contained both pressure and particle motion components. Sound pressure and particle acceleration thresholds varied significantly among species and between frequencies; audiograms were notably flatter for acceleration than pressure at low frequencies. Thresholds of species with diverticulae projecting anteriorly from their swim bladders ( weakfish, spotted seatrout, and Atlantic croaker) were typically but not significantly lower than those of species lacking such projections ( red drum, spot, northern kingfish). Sciaenids were most sensitive at low frequencies that overlap the peak frequencies of their vocalizations. Auditory thresholds of these species were used to estimate idealized propagation distances of sciaenid vocalizations in coastal and estuarine environments

Enhanced H2O2 Production via Photocatalytic O2 Reduction over Structurally-Modified Poly(heptazine imide)

Solar H2O2 produced by O2 reduction provides a green, efficient, and ecological alternative to the industrial anthraquinone process and H2/O2 direct-synthesis. We report efficient photocatalytic H2O2 production at a rate of 73.4 mM h–1 in the presence of a sacrificial donor on a structurally engineered catalyst, alkali metal-halide modulated poly(heptazine imide) (MX → PHI). The reported H2O2 production is nearly 150 and >4250 times higher than triazine structured pristine carbon nitride under UV–visible and visible light (≥400 nm) irradiation, respectively. Furthermore, the solar H2O2 production rate on MX → PHI is higher than most of the previously reported carbon nitride (triazine, tri-s-triazine), metal oxides, metal sulfides, and other metal–organic photocatalysts. A record high AQY of 96% at 365 nm and 21% at 450 nm was observed. We find that structural modulation by alkali metal-halides results in a highly photoactive MX → PHI catalyst which has a broader light absorption range, enhanced light absorption ability, tailored bandgap, and a tunable band edge position. Moreover, this material has a different polymeric structure, high O2 trapping ability, interlayer intercalation, as well as surface decoration of alkali metals. The specific C≡N groups and surface defects, generated by intercalated MX, were also considered as potential contributors to the separation of photoinduced electron–hole pairs, leading to enhanced photocatalytic activity. A synergy of all these factors contributes to a higher H2O2 production rate. Spectroscopic data help us to rationalize the exceptional photochemical performance and structural characteristics of MX → PHI

Cd/Pt Precursor Solution for Solar H-2 Production and in situ Photochemical Synthesis of Pt Single-atom Decorated CdS Nanoparticles

Despite extensive efforts to develop high-performance H2 evolution catalysts, this remains a major challenge. Here, we demonstrate the use of Cd/Pt precursor solutions for significant photocatalytic H2 production (154.7 mmol g−1 h−1), removing the need for a pre-synthesized photocatalyst. In addition, we also report simultaneous in situ synthesis of Pt single-atoms anchored CdS nanoparticles (PtSA-CdSIS) during photoirradiation. The highly dispersed in situ incorporation of extensive Pt single atoms on CdSIS enables the enhancement of active sites and suppresses charge recombination, which results in exceptionally high solar-to-hydrogen conversion efficiency of ≈1 % and an apparent quantum yield of over 91 % (365 nm) for H2 production. Our work not only provides a promising strategy for maximising H2 production efficiency but also provides a green process for H2 production and the synthesis of highly photoactive PtSA-CdSIS nanoparticles

Does self-monitoring reduce blood pressure? Meta-analysis with meta-regression of randomized controlled trials

Introduction. Self-monitoring of blood pressure (BP) is an increasingly common part of hypertension management. The objectives of this systematic review were to evaluate the systolic and diastolic BP reduction, and achievement of target BP, associated with self-monitoring. Methods. MEDLINE, Embase, Cochrane database of systematic reviews, database of abstracts of clinical effectiveness, the health technology assessment database, the NHS economic evaluation database, and the TRIP database were searched for studies where the intervention included self-monitoring of BP and the outcome was change in office/ambulatory BP or proportion with controlled BP. Two reviewers independently extracted data. Meta-analysis using a random effects model was combined with meta-regression to investigate heterogeneity in effect sizes. Results. A total of 25 eligible randomized controlled trials (RCTs) (27 comparisons) were identified. Office systolic BP (20 RCTs, 21 comparisons, 5,898 patients) and diastolic BP (23 RCTs, 25 comparisons, 6,038 patients) were significantly reduced in those who self-monitored compared to usual care (weighted mean difference (WMD) systolic −3.82 mmHg (95% confidence interval −5.61 to −2.03), diastolic −1.45 mmHg (−1.95 to −0.94)). Self-monitoring increased the chance of meeting office BP targets (12 RCTs, 13 comparisons, 2,260 patients, relative risk = 1.09 (1.02 to 1.16)). There was significant heterogeneity between studies for all three comparisons, which could be partially accounted for by the use of additional co-interventions. Conclusion. Self-monitoring reduces blood pressure by a small but significant amount. Meta-regression could only account for part of the observed heterogeneity