Regulation of Type IV Pili Contributes to Surface Behaviors of Historical and Epidemic Strains of Clostridium difficile

ABSTRACT The intestinal pathogen Clostridium difficile is an urgent public health threat that causes antibiotic-associated diarrhea and is a leading cause of fatal nosocomial infections in the United States. C. difficile rates of recurrence and mortality have increased in recent years due to the emergence of so-called “hypervirulent” epidemic strains. A great deal of the basic biology of C. difficile has not been characterized. Recent findings that flagellar motility, toxin synthesis, and type IV pilus (TFP) formation are regulated by cyclic diguanylate (c-di-GMP) reveal the importance of this second messenger for C. difficile gene regulation. However, the function(s) of TFP in C. difficile remains largely unknown. Here, we examine TFP-dependent phenotypes and the role of c-di-GMP in controlling TFP production in the historical 630 and epidemic R20291 strains of C. difficile . We demonstrate that TFP contribute to C. difficile biofilm formation in both strains, but with a more prominent role in R20291. Moreover, we report that R20291 is capable of TFP-dependent surface motility, which has not previously been described in C. difficile . The expression and regulation of the pilA1 pilin gene differs between R20291 and 630, which may underlie the observed differences in TFP-mediated phenotypes. The differences in pilA1 expression are attributable to greater promoter-driven transcription in R20291. In addition, R20291, but not 630, upregulates c-di-GMP levels during surface-associated growth, suggesting that the bacterium senses its substratum. The differential regulation of surface behaviors in historical and epidemic C. difficile strains may contribute to the different infection outcomes presented by these strains. IMPORTANCE How Clostridium difficile establishes and maintains colonization of the host bowel is poorly understood. Surface behaviors of C. difficile are likely relevant during infection, representing possible interactions between the bacterium and the intestinal environment. Pili mediate bacterial interactions with various surfaces and contribute to the virulence of many pathogens. We report that type IV pili (TFP) contribute to biofilm formation by C. difficile . TFP are also required for surface motility, which has not previously been demonstrated for C. difficile . Furthermore, an epidemic-associated C. difficile strain showed higher pilin gene expression and greater dependence on TFP for biofilm production and surface motility. Differences in TFP regulation and their effects on surface behaviors may contribute to increased virulence in recent epidemic strains

Code density concerns for new architectures

Reducing a program\u27s instruction count can improve cache behavior and bandwidth utilization, lower power consumption, and increase overall performance. Nonetheless, code density is an often overlooked feature in studying processor architectures. We hand-optimize an assembly language embedded benchmark for size on 21 different instruction set architectures, finding up to a factor of three difference in code sizes from ISA alone. We find that the architectural features that contribute most heavily to code density are instruction length, number of registers, availability of a zero register, bit-width, hardware divide units, number of instruction operands, and the availability of unaligned loads and stores. We extend our results to investigate operating system, compiler, and system library effects on code density. We find that the executable starting address, executable format, and system call interface all affect program size. While ISA effects are important, the efficiency of the entire system stack must be taken into account when developing a new dense instruction set architecture

Examining the Lived Experience of Mild Cognitive Impairment: An Evidence-Based Practice Project

This Evidence-Based Practice (EBP) project addressed the following question: What are the perspectives, experiences, and self-reports of adult individuals, groups, or populations who have MCI or report problems with Functional Cognition (and / or their caregivers)

Pulsar scintillation through thick and thin: Bow shocks, bubbles, and the broader interstellar medium

Observations of pulsar scintillation are among the few astrophysical probes of very small-scale (≲ au) phenomena in the interstellar medium (ISM). In particular, characterization of scintillation arcs, including their curvature and intensity distributions, can be related to interstellar turbulence and potentially overpressurized plasma in local ISM inhomogeneities, such as supernova remnants, H II regions, and bow shocks. Here we present a survey of eight pulsars conducted at the Five-hundred-metre Aperture Spherical Telescope (FAST), revealing a diverse range of scintillation arc characteristics at high sensitivity. These observations reveal more arcs than measured previously for our sample. At least nine arcs are observed toward B1929+10 at screen distances spanning ~90 per cent of the pulsar’s 361 pc path length to the observer. Four arcs are observed toward B0355+54, with one arc yielding a screen distance as close as ∼105 au (<1 pc) from either the pulsar or the observer. Several pulsars show highly truncated, low-curvature arcs that may be attributable to scattering near the pulsar. The scattering screen constraints are synthesized with continuum maps of the local ISM and other well-characterized pulsar scintillation arcs, yielding a three-dimensional view of the scattering media in context

Multiple enzymatic approaches to hydrolysis of fungal beta-glucans by the soil bacterium Chitinophaga pinensis

The genome of the soil Bacteroidota Chitinophaga pinensis encodes a large number of glycoside hydrolases (GHs) with noteworthy features and potentially novel functions. Several are predicted to be active on polysaccharide components of fungal and oomycete cell walls, such as chitin, beta-1,3-glucan and beta-1,6-glucan. While several fungal beta-1,6-glucanase enzymes are known, relatively few bacterial examples have been characterised to date. We have previously demonstrated that C. pinensis shows strong growth using beta-1,6-glucan as the sole carbon source, with the efficient release of oligosaccharides from the polymer. We here characterise the capacity of the C. pinensis secretome to hydrolyse the beta-1,6-glucan pustulan and describe three distinct enzymes encoded by its genome, all of which show different levels of beta-1,6-glucanase activity and which are classified into different GH families. Our data show that C. pinensis has multiple tools to deconstruct pustulan, allowing the species' broad utility of this substrate, with potential implications for bacterial biocontrol of pathogens via cell wall disruption. Oligosaccharides derived from fungal beta-1,6-glucans are valuable in biomedical research and drug synthesis, and these enzymes could be useful tools for releasing such molecules from microbial biomass, an underexploited source of complex carbohydrates

Ditching the Driving: A Cross-Sectional Study on the Determinants of Remote Work From Home for Sign Language Interpreters.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic dramatically impacted the working conditions for sign language interpreters, shifting the provision of interpreting services from onsite to remote. The goal of this cross-sectional study was to examine the perceptions of determinants of remote interpreting implementation from home by sign language interpreters during the pandemic. We hypothesized that interpreters working across the primary settings of staff (agency, government, business, or hospital employees), educational (K-12 or postsecondary), community/freelance (independent contractor), video remote (the two-way connection between onsite participants and remote interpreter), and video relay (three-way telecommunication) would present with differing experiences of the implementation process. METHODS: The Determinants of Implementation Behavior Questionnaire was adapted for sign language interpreters (DIBQ-SLI) and administered to certified interpreters working remotely at least 10 h per week. The DIBQ-SLI included eight constructs (knowledge, skills, self-efficacy, perceived behavioral control, innovation characteristics, organizational resources and support, innovation strategies, and social support) and 30 items. Parametric statistics assessed differences in interpreters' perceptions across settings. Principal component analysis was conducted for data reduction and affirmation of the most critical constructs and items. RESULTS: One hundred and six interpreters (37 video relay, 27 video remote, 18 educational, 11 community/freelance, 11 staff interpreters, and two from "other" settings) completed the DIBQ-SLI. The video relay and staff interpreters consistently demonstrated the most favorable and the educational interpreters demonstrated the least favorable perceptions. Of the total variance, 58.8% of interpreters' perceptions was explained by organizational (41%), individual (10.7%), and social (7.1%) dimensions. There were significant differences across settings for the organizational and individual principal components; however, no differences were detected for the social principal component. CONCLUSIONS: An administrative infrastructure devoted to ensuring that interpreters receive sufficient managerial support, training, materials and resources, experience with remote interpreting before having to commit, and insights based on the results of their remote work (organizational principal component) may be necessary for improving perceptions. Remote interpreting is expected to continue after the pandemic ends; thus, settings with the least favorable ratings across behavior constructs may borrow strategies from settings with the most favorable ratings to help promote perceptions of the contextual determinants of future remote interpreting implementation

The chronostratigraphy of Late Pleistocene glacial and periglacial aeolian activity in the Tuktoyaktuk Coastlands, NWT, Canada

Aeolian periglacial sand deposits are common in the Tuktoyaktuk Coastlands of Western Arctic Canada. Regionally extensive and thick aeolian sand-sheet deposits have been observed in two major stratigraphic settings: within a sand unit characterized by large aeolian dune deposits; and interbedded with glaciofluvial outwash from the Laurentide Ice Sheet (LIS). Small, localized sand sheets have also been observed along the tops of sandy bluffs, within sequences of drained thermokarst lakes deposits and as an involuted veneer above buried basal ice of the LIS. On the basis of radiocarbon and optically stimulated luminescence (OSL) dates from preserved periglacial aeolian sand sheets and dunes a regional chronostratigraphy is presented which indicates that both extensive dunes and sand sheets accumulated mainly between ca 30 and 13 ka. A switch to dominantly sand-sheet aggradation at ca 14–13 ka, with sand sheets forming widely until ca 8 ka, is attributed to (a) surface armouring by glacial deposits associated with the advance of the LIS; and (b) amelioration of the climate from cold aridity. An absence of OSL dates between ca 8 and 1 ka suggests that sand sheets stabilized during much of the Holocene. Local sand-sheet aggradation during recent centuries has occurred near sandy bluffs and on the floors of drained thermokarst lakes. The OSL dates constrain the maximum extent of the LIS in the Tuktoyaktuk Coastlands to Marine Isotope Stage 2

Glycan processing in gut microbiomes

Microbiomes and their enzymes process many of the nutrients accessible in the gastrointestinal tract of bilaterians and play an essential role in host health and nutrition. In this review, we describe recent insights into nutrient processing in microbiomes across three exemplary yet contrasting gastrointestinal ecosystems (humans, ruminants and insects), with focus on bacterial mechanisms for the utilization of common and atypical dietary glycans as well as host-derived mucus glycans. In parallel, we discuss findings from multi-omic studies that have provided new perspectives on understanding glycan-dependent interactions and the complex food-webs of microbial populations in their natural habitat. Using key examples, we emphasize how increasing understanding of glycan processing by gut microbiomes can provide critical insights to assist ‘microbiome reprogramming’, a growing field that seeks to leverage diet to improve animal growth and host health