A novel NGS library preparation method to characterize native termini of fragmented DNA.

Biological and chemical DNA fragmentation generates DNA molecules with a variety of termini, including blunt ends and single-stranded overhangs. We have developed a Next Generation Sequencing (NGS) assay, XACTLY, to interrogate the termini of fragmented DNA, information traditionally lost in standard NGS library preparation methods. Here we describe the XACTLY method, showcase its sensitivity and specificity, and demonstrate its utility in in vitro experiments. The XACTLY assay is able to report relative abundances of all lengths and types (5' and 3') of single-stranded overhangs, if present, on each DNA fragment with an overall accuracy between 80-90%. In addition, XACTLY retains the sequence of each native DNA molecule after fragmentation and can capture the genomic landscape of cleavage events at single nucleotide resolution. The XACTLY assay can be applied as a novel research and discovery tool for fragmentation analyses and in cell-free DNA

Sex Differences in Arm Muscle Fatigability With Cognitive Demand in Older Adults

Background Muscle fatigability can increase when a stressful, cognitively demanding task is imposed during a low-force fatiguing contraction with the arm muscles, especially in women. Whether this occurs among older adults (\u3e 60 years) is currently unknown. Questions/purposes We aimed to determine if higher cognitive demands, stratified by sex, increased fatigability in older adults (\u3e 60 years). Secondarily, we assessed if varying cognitive demand resulted in decreased steadiness and was explained by anxiety or cortisol levels. Methods Seventeen older women (70 ± 6 years) and 13 older men (71 ± 5 years) performed a sustained, isometric, fatiguing contraction at 20% of maximal voluntary contraction until task failure during three sessions: high cognitive demand (high CD = mental subtraction by 13); low cognitive demand (low CD = mental subtraction by 1); and control (no subtraction). Results Fatigability was greater when high and low CD were performed during the fatiguing contraction for the women but not for the men. In women, time to failure with high CD was 16 ± 8 minutes and with low CD was 17 ± 4 minutes, both of which were shorter than time to failure in control contractions (21 ± 7 minutes; high CD mean difference: 5 minutes [95% confidence interval {CI}, 0.78–9.89], p = 0.02; low CD mean difference: 4 minutes [95% CI, 0.57–7.31], p = 0.03). However, in men, no differences were detected in time to failure with cognitive demand (control: 13 ± 5 minutes; high CD mean difference: −0.09 minutes [95% CI, −2.8 to 2.7], p = 1.00; low CD mean difference: 0.75 minutes [95% CI, −1.1 to 2.6], p = 0.85). Steadiness decreased (force fluctuations increased) more during high CD than control. Elevated anxiety, mean arterial pressure, and salivary cortisol levels in both men and women did not explain the greater fatigability during high CD. Conclusions Older women but not men showed marked increases in fatigability when low or high CD was imposed during sustained static contractions with the elbow flexor muscles and contrasts with previous findings for the lower limb. Steadiness decreased in both sexes when high CD was imposed. Clinical Relevance Older women are susceptible to greater fatigability of the upper limb with heightened mental activity during sustained postural contractions, which are the foundation of many work-related tasks

Liquid-filled hard gelatin capsules : excipient/capsule compatibility studies

Encapsulation of pharmaceutical formulations as liquids or semisolids, within hard gelatin capsules, presents an important oral dosage strategy for poorly water-soluble drugs, resulting in good bioavailability and reproducible drug absorption. In addition, this technology offers an inherently safer process than powder filled capsules and tablets for highly potent or cytotoxic drugs by avoiding dust generation. Here we present a compatibility study of hard gelatin capsules with common excipients in absence of active pharmaceutical

Stress corrosion in titanium alloys and other metallic materials

Multiple physical and chemical techniques including mass spectroscopy, atomic absorption spectroscopy, gas chromatography, electron microscopy, optical microscopy, electronic spectroscopy for chemical analysis (ESCA), infrared spectroscopy, nuclear magnetic resonance (NMR), X-ray analysis, conductivity, and isotopic labeling were used in investigating the atomic interactions between organic environments and titanium and titanium oxide surfaces. Key anhydrous environments studied included alcohols, which contain hydrogen; carbon tetrachloride, which does not contain hydrogen; and mixtures of alcohols and halocarbons. Effects of dissolved salts in alcohols were also studied. This program emphasized experiments designed to delineate the conditions necessary rather than sufficient for initiation processes and for propagation processes in Ti SCC

Towards a consistent mechanism of emulsion polymerization—new experimental details

The application of atypical experimental methods such as conductivity measurements, optical microscopy, and nonstirred polymerizations to investigations of the ‘classical’ batch ab initio emulsion polymerization of styrene revealed astonishing facts. The most important result is the discovery of spontaneous emulsification leading to monomer droplets even in the quiescent styrene in water system. These monomer droplets with a size between a few and some hundreds of nanometers, which are formed by spontaneous emulsification as soon as styrene and water are brought into contact, have a strong influence on the particle nucleation, the particle morphology, and the swelling of the particles. Experimental results confirm that micelles of low-molecular-weight surfactants are not a major locus of particle nucleation. Brownian dynamics simulations show that the capture of matter by the particles strongly depends on the polymer volume fraction and the size of the captured species (primary free radicals, oligomers, single monomer molecules, or clusters)

The integrated academic information system support for education 3.0 in higher education institution: lecturer perspective

Education 3.0 has been implemented in many higher education institutions (HEIs). Education 3.0 has been directed the institution toward better educational experience. But on the other hands, the implementation of Education 3.0 also caused some problems. Previous research has found administrative problem experienced by the lecturer. This research explores deeper from the lecturer and suggested the solution from lecturer perspective, combined with information technology capabilities owned by the HEIs. The research used a case study as the method and conducted a qualitative research with a semi-structured interview. The interview analysis has found that the increase of the administrative processes is caused by online and offline administrative activities. The online activities are from e-learning and the offline activities are from traditional learning (face-to-face). The administrative processes also involved the academic information system (AIS). Simplified all of the administrative processes are more preferred. To overcome the problems, integrating the AIS and e-learning become necessary. This research suggests transforming the existing AIS into an integrated AIS and hopes the solution can simplify the administration process

Geographically dispersed zoonotic tuberculosis in pre-contact South American human populations

Previous ancient DNA research has shown that Mycobacterium pinnipedii, which today causestuberculosis (TB) primarily in pinnipeds, infected human populations living in the coastalareas of Peru prior to European colonization. Skeletal evidence indicates the presence of TB inseveral pre-colonial South and North American populations with minimal access to marineresources— a scenario incompatible with TB transmission directly from infected pinnipeds ortheir tissues. In this study, we investigate the causative agent of TB in ten pre-colonial, non-coastal individuals from South America. We reconstruct M. pinnipedii genomes (10- to 15-foldmean coverage) from three contemporaneous individuals from inland Peru and Colombia,demonstrating the widespread dissemination of M. pinnipedii beyond the coast, either throughhuman-to-human and/or animal-mediated routes. Overall, our study suggests that TBtransmission in the pre-colonial era Americas involved a more complex transmission pathwaythan simple pinniped-to-human transfer

The widespread use of topical antimicrobials enriches for resistance in Staphylococcus aureus isolated from patients with atopic dermatitis.

BACKGROUND: Carriage rates of Staphylococcus aureus on affected skin in atopic dermatitis (AD) are approximately 70%. Increasing disease severity during flares and overall disease severity correlate with increased burden of S. aureus. Treatment in AD therefore often targets S. aureus with topical and systemic antimicrobials. OBJECTIVES: To determine whether antimicrobial sensitivities and genetic determinants of resistance differed in S. aureus isolates from the skin of children with AD and healthy child nasal carriers. METHODS: In this case-control study, we compared S. aureus isolates from children with AD (n = 50) attending a hospital dermatology department against nasal carriage isolates from children without skin disease (n = 49) attending a hospital emergency department for noninfective conditions. Using whole genome sequencing we generated a phylogenetic framework for the isolates based on variation in the core genome, then compared antimicrobial resistance phenotypes and genotypes between disease groups. RESULTS: Staphylococcus aureus from cases and controls had on average similar numbers of phenotypic resistances per isolate. Case isolates differed in their resistance patterns, with fusidic acid resistance (FusR ) being significantly more frequent in AD (P = 0·009). The genetic basis of FusR also differentiated the populations, with chromosomal mutations in fusA predominating in AD (P = 0·049). Analysis revealed that FusR evolved multiple times and via multiple mechanism in the population. Carriage of plasmid-derived qac genes, which have been associated with reduced susceptibility to antiseptics, was eight times more frequent in AD (P = 0·016). CONCLUSIONS: The results suggest that strong selective pressure drives the emergence and maintenance of specific resistances in AD