Explore the vast range of titles available.

Introduction Reductions in fibrinolytic potential occur with both aging and physical inactivity and are associated with an increased cardiovascular disease risk. Plasmin, the enzyme responsible for the enzymatic degradation of fibrin clots, is activated by tissue plasminogen activator (tPA), while plasminogen activator inhibitor-1 (PAI-1) inhibits its activation. Currently, fibrinolysis research focuses almost exclusively on changes within the plasma. However, tPA and PAI-1 are expressed by human skeletal muscle (SM). Currently, no studies have focused on changes in SM fibrinolytic activity with regard to aging and aerobic fitness. Purpose The purpose of this study was to cross-sectionally evaluate effects of age and aerobic fitness on tPA and PAI-1 expressions and activity in SM. Methods Twenty-six male subjects were categorized into the following groups: (1) young aerobically trained ( n  = 8); (2) older aerobically trained ( n  = 6); (3) young aerobically untrained ( n  = 7); and (4) older aerobically untrained ( n  = 5). Muscle biopsies were obtained from each subject. SM tPA activity was assessed using gel zymography and SM tPA and PAI-1 expressions were assessed using RT-PCR. Results Trained subjects had higher SM tPA activity compared to untrained (25.3 ± 2.4 × 10 3 vs. 21.5 ± 5.6 × 10 3  pixels, respectively; p  = 0.03) with no effect observed for age. VO 2 max and SM tPA activity were also significantly correlated ( r  = 0.42; p  < 0.04). SM tPA expression was higher in older participants, but no effect of fitness level was observed. No differences were observed for PAI-1 expression in SM. Conclusions Higher levels of aerobic fitness are associated with increased fibrinolytic activity in SM.

Hydrogel biomaterials have proven indispensable for three-dimensional (3D) cell culture but have fallen short in replicating the innate physiochemical complexity of real tissue. Though traditional photolithography affords localized material manipulation, robust methods that govern when, where, and to what extent such phototailoring occurs throughout materials would be profoundly enabling towards fabricating more-realistic 3D tissue constructs. Here, we introduce \"grayscale image z-stack-guided multiphoton optical-lithography\" (GIZMO) as a generalizable and intuitive strategy to rapidly photomodulate materials in full 3D non-binary patterns at submicron resolutions spanning large volumes (>mm3). Highlighting its versatility, we employ GIZMO to variably photopattern biomolecule release from, protein immobilization to, and degradation within hydrogels based on biologically derived or synthetic grayscale image stacks with unprecedented complexity. We anticipate that GIZMO will enable new opportunities to probe and manipulate cell fates, as well as to engineer complex functional tissue.Competing Interest StatementG.J. is an employee of MBF Bioscience, the developer of ScanImage. C.A.D. is an inventor on a related provisional patent application submitted by the University of Washington. All other authors declare no competing financial interests.

Abstract A major challenge in synthetic biology is properly balancing evolved and engineered functions without compromising microbial fitness. Many microbial proteins are not required for growth in regular laboratory conditions, but it is unclear what fraction of the proteome can be eliminated to increase bioproduction and maintain fitness. Here, we investigated the effects of massive genome reduction in E. coli on the expression level and evolutionary stability of a model biosynthetic pathway to produce the pigment protodeoxyviolacein (PDV). We identified an amino acid metabolism imbalance and compromised growth that were correlated with elimination of genes associated with significant proteome fraction. Proteomic profiling suggested that increased amino acid pools are responsible for an alleviation of fitness defects associated with PDV expression. In addition, all strains with genome reductions that significantly affected the proteome exhibited decreased stability of PDV production compared to the wild-type strain under persistent PDV expression conditions despite the alleviation of fitness defects. These findings exhibit the importance of balancing evolved functions with engineered ones to achieve an optimal balance of fitness and bioproduction. Competing Interest Statement The authors have declared no competing interest.

Genetic screens are essential for uncovering novel molecular mechanisms and identifying the functions of hypothetical proteins. CRISPR interference (CRISPRi) is a powerful, programmable, and sequence-specific gene repression technology that can be used for high-throughput screening and targeted gene repression. Despite its ease of use, the initial development of CRISPRi systems is labor-intensive in many non-model organisms. Our goal is to simplify this by establishing a host-agnostic CRISPRi platform that utilizes the serine recombinase-assisted genome engineering (SAGE) system. This system integrates CRISPRi machinery directly into the bacterial chromosome, overcoming the limitations of plasmid-based systems and enabling wide sharing across diverse bacteria. We demonstrate the design and optimization of multiplexed CRISPRi to repress multiple genes simultaneously in phylogenetically distant bacteria. We use a Francisella novicida-derived Cas12a system that processes multiple distinct CRISPR RNAs, each targeting a unique gene sequence, from a single transcript. This allows easy multi-gene repression. By reinforcing gene repression with multiple guides targeting a single gene, we achieve robust genetic perturbations without the need to pre-screen the efficacy of guide RNAs. Using this toolkit, we perturb multiple combinations of growth and visual phenotypes in Pseudomonas fluorescens and demonstrate simultaneous repression of multiple fluorescent proteins to near background levels in bacteria from various other genera. While the tools are directly portable to all SAGE-compatible microbes, we illustrate the utility of SAGE by optimizing CRISPRi performance in Rhodococcus jostii through a combinatorial screen of Cas protein and CRISPR array expression variants. The efficient integration of CRISPRi machinery via the SAGE system paves the way for versatile genetic screening, enabling profound insights into gene functions both in laboratory conditions and relevant naturalistic scenarios.

Ryan recounts his summer camp experience as the helper for Max, the camp maintenance man. Max, who loved \"his\" camp, enjoyed explaining the working of things to his young assisstant.

Climate drives population dynamics through multiple mechanisms, which can lead to seemingly context-dependent effects of climate on natural populations. For climate-sensitive diseases such as dengue, chikungunya, and Zika, climate appears to have opposing effects in different contexts. Here we show that a model, parameterized with laboratory measured climate-driven mosquito physiology, captures three key epidemic characteristics across ecologically and culturally distinct settings in Ecuador and Kenya: the number, timing, and duration of outbreaks. The model generates a range of disease dynamics consistent with observed Aedes aegypti abundances and laboratory-confirmed arboviral incidence with variable accuracy (28–85% for vectors, 44–88%for incidence). The model predicted vector dynamics better in sites with a smaller proportion of young children in the population, lower mean temperature, and homes with piped water and made of cement. Models with limited calibration that robustly capture climate-virus relationships can help guide intervention efforts and climate change disease projections.

Versatile and precise genome modifications are needed to create a wider range of adoptive cellular therapies 1 – 5 . Here we report two improvements that increase the efficiency of CRISPR–Cas9-based genome editing in clinically relevant primary cell types. Truncated Cas9 target sequences (tCTSs) added at the ends of the homology-directed repair (HDR) template interact with Cas9 ribonucleoproteins (RNPs) to shuttle the template to the nucleus, enhancing HDR efficiency approximately two- to fourfold. Furthermore, stabilizing Cas9 RNPs into nanoparticles with polyglutamic acid further improves editing efficiency by approximately twofold, reduces toxicity, and enables lyophilized storage without loss of activity. Combining the two improvements increases gene targeting efficiency even at reduced HDR template doses, yielding approximately two to six times as many viable edited cells across multiple genomic loci in diverse cell types, such as bulk (CD3 + ) T cells, CD8 + T cells, CD4 + T cells, regulatory T cells (Tregs), γδ T cells, B cells, natural killer cells, and primary and induced pluripotent stem cell-derived 6 hematopoietic stem progenitor cells (HSPCs). Precise genome editing is made more efficient by stabilizing Cas9 and enhancing shuttling to the nucleus.

Wind turbines have been hypothesized to affect bat populations; however, no comprehensive analysis of bat mortality from the operation of wind turbines in Canada has been conducted. We used data from carcass searches for 64 wind farms, incorporating correction factors for scavenger removal, searcher efficiency, and carcasses that fell beyond the area searched to estimate bat collision mortality associated with wind turbines in Canada. On average, 15.5±3.8 (95% CI) bats were killed per turbine per year at these sites (range=0-103 bats/turbine/yr at individual wind farms). Based on 4,019 installed turbines (the no. installed in Canada by Dec 2013), an estimated 47,400 bats (95% CI=32,100-62,700) are killed by wind turbines each year in Canada. Installed wind capacity is growing rapidly in Canada, and is predicted to increase approximately 3.5-fold over the next 15 years, which could lead to direct mortality of approximately 166,000 bats/year. Long-distance migratory bat species (e.g., hoary bat [Lasiurus cinereus], silver-haired bat [Lasionycteris noctivagans], eastern red bat [Lasiurus borealis]) accounted for 73% of all mortalities. These species are subject to additional mortality risks when they migrate into the United States. The little brown myotis (Myotis lucifugus), which was listed as Endangered in 2014 under the Species At Risk Act (SARA), accounted for 13% of all mortalities from wind turbines, with most of the mortality (87%) occurring in Ontario. Population-level impacts may become an issue for some bat species as numbers of turbines increase. © 2016 The Wildlife Society.

Just as geological samples from Earth record the natural history of our planet, astromaterials hold the natural history of our solar system and beyond. Astromaterials acquisition and curation practices have direct consequences on the contamination levels of astromaterials and hence the types of questions that can be answered about our solar system and the degree of precision that can be expected of those answers. Advanced curation was developed as a cross-disciplinary field to improve curation and acquisition practices in existing astromaterials collections and for future sample return activities, including meteorite and cosmic dust samples that are collected on Earth. These goals are accomplished through research and development of new innovative technologies and techniques for sample collection, handling, characterization, analysis, and curation of astromaterials. In this contribution, we discuss five broad topics in advanced curation that are critical to improving sample acquisition and curation practices, including (1) best practices for monitoring and testing of curation infrastructure for inorganic, organic, and biological contamination; (2) requirements for storage, processing, and sample handling capabilities for future sample return missions, along with recent progress in these areas; (3) advancements and improvements in astromaterials acquisition capabilities on Earth (i.e., the collection of meteorites and cosmic dust); (4) the importance of contamination knowledge strategies for maximizing the science returns of sample-return missions; and (5) best practices and emerging capabilities for the basic characterization and preliminary examination of astromaterials. The primary result of advanced curation research is to both reduce and quantify contamination of astromaterials and preserve the scientific integrity of all samples from mission inception to secure delivery of samples to Earth-based laboratories for in-depth scientific analysis. Advanced curation serves as an important science-enabling activity, and the collective lessons learned from previous spacecraft missions and the results of advanced curation research will work in tandem to feed forward into better spacecraft designs and enable more stringent requirements for future sample return missions and Earth-based sample acquisition.