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Osteocytes are the resident mechanosensory cells in bone. They are responsible for skeletal homeostasis and adaptation to mechanical cues. Integrin proteins play an prominent role in osteocyte mechanotransduction, however the details are not well stratifiedin vivo. Intravital imaging with multiphoton microscopy presents an opportunity to study molecular level mechanobiological eventsin vivo, and could be used to study integrin dynamics in osteocytes. However, fluorescent imaging limitations with respect to excessive optical scattering and low signal to noise ratio caused by mineralized bone matrix make such investigations non-trivial. Here we demonstrate that ultra-small and bright fluorescent core-shell silica nanoparticles (<7nm diameter), known as Cornell Prime Dots (C’Dots), are well-suited for thein vivobone microenvironment and can improve intravital imaging capabilities. We report validation studies for C’Dots as a novel, locally injectedin vivoosteocyte imaging tool for both non-specific cellular uptake and for targeting integrins. The pharmacokinetics of C’Dots reveal distinct sex differences in nanoparticle cycling and clearance in osteocytes, which represents a novel topic of study in bone biology. Integrin-targeted C’Dots were used to study osteocyte integrin dynamics. To the best of our knowledge, we report here the first evidence of osteocyte integrin endocytosis and recyclingin vivo. Our results provide novel insights in osteocyte biology and will open up new lines of investigation that were previously unavailablein vivo.

Marsupials display accelerated development of the craniofacial region relative to the neurocranium when compared to placental mammals. This is thought to facilitate suckling by the highly altricial neonate after making the journey into the pouch. Whilecis-regulatory regions are considered to play a significant role in morphological evolution the face, the genetic mechanisms involved in craniofacial heterochrony among the major mammal lineages remain unclear. Here, we compared thecis-regulatory landscapes of the fat-tailed dunnart (Sminthopsis crassicaudata; Dasyuridae), a small marsupial amenable to laboratory husbandry, and mouse to gain insights into the basis of heterochrony. We compared patterns of the chromatin modifications, H3K4me3 and H3K27ac, between the dunnart and mouse during developmental stages when homologous craniofacial structures form each in species. We found that dunnart promoter- and enhancer-associated peaks at the time of birth in the dunnart broadly overlapped with all the stages of embryonic craniofacial development assessed in the mouse. However, dunnart-specific peaks were significantly enriched around genes whose mouse orthologs exhibit increased expression in the face over time. Moreover, genes displaying this temporal expression pattern were enriched for Gene Ontology terms related to ossification and skeletal development, processes that underlie development of the cranial muscles and bones of the face. This suggests a greater similarity between immediate postnatal chromatin landscape in the dunnart and late embryonic craniofacial development in the mouse. Using mouse-dunnart comparisons, we also discovered evidence of dunnart-specific peaks active near genes involved in the development of mechanosensory structures that may relate to the distinctive postnatal journey marsupial young take to the reach the pouch. This study characterisedcis-regulatory elements driving craniofacial development in marsupials and their potential role in craniofacial heterochrony.

Recent advances in long-read sequencing technology have allowed for single-molecule sequencing of entire mitochondrial genomes, opening the door for direct investigation of the mitochondrial genome architecture and recombination. We used PacBio sequencing to reassemble mitochondrial genomes from two species of New Zealand freshwater snails, Potamopyrgus antipodarum and Potamopyrgus estuarinus. These assemblies revealed a ∼1.7 kb structure within the mitochondrial genomes of both species that was previously undetected by an assembly of short reads and likely corresponding to a large noncoding region commonly present in the mitochondrial genomes. The overall architecture of these Potamopyrgus mitochondrial genomes is reminiscent of the chloroplast genomes of land plants, harboring a large single-copy (LSC) region and a small single-copy (SSC) region separated by a pair of inverted repeats (IRa and IRb). Individual sequencing reads that spanned across the Potamopyrgus IRa-SSC-IRb structure revealed the occurrence of a “flip-flop” recombination. We also detected evidence for two distinct IR haplotypes and recombination between them in wild-caught P. estuarinus, as well as extensive intermolecular recombination between single-nucleotide polymorphisms in the LSC region. The chloroplast-like architecture and repeat-mediated mitochondrial recombination we describe here raise fundamental questions regarding the origins and commonness of inverted repeats in cytoplasmic genomes and their role in mitochondrial genome evolution.

Forest harvesting machines have become technologically advanced, making them more productive but also more complex and expensive to purchase. Logging contractors have the challenge of deciding the ...

Scholarship on human rights and environmental justice suggests that organizations vary in their messaging regarding outcomes related to environmental protection and sustainability, differences often found in the divide between the Global North and Global South. The literature also suggests that Indigenous organizations represent groups that traditionally focus on issues of sovereignty, while grappling with unique problems related to assimilation, cultural preservation, and oppression. This study utilizes empirical data gathered from 333 non-governmental organizations affiliated with the United Nations Human Rights Council to explore whether Indigenous and non-Indigenous organizations, which share many aspects of their mission with one another at the transnational level, differ on issues related to environment sustainability and collective identity rights. Our results indicate that Indigenous organizations take a more holistic approach in addressing the relationship between humans and the natural world, centring marginalized perspectives through restorative justice and the needs of current and future generations.

Recent advances in long-read sequencing technology have allowed for single-molecule sequencing of entire mitochondrial genomes, opening the door for direct investigation of mitochondrial genome architecture and landscapes of recombination. We used PacBio sequencing to re-assemble mitochondrial genomes from two species of New Zealand freshwater snails, Potamopyrgus antipodarum and Potamopyrgus estuarinus. These assemblies revealed a ∼1.7 kb structure within the mitochondrial genomes of both species that was previously undetected by assembly of short sequencing reads and likely corresponding to a large non-coding region commonly present in mitochondrial genomes. The overall architecture of these Potamopyrgus mitochondrial genomes is reminiscent of the chloroplast genomes of land plants, harboring a large single-copy region (LSC) and a small single-copy region (SSC) separated by a pair of inverted repeats (IRa and IRb). Individual sequencing reads that spanned across the Potamopyrgus IRa–SSC–IRb structure revealed the occurrence of “flip-flop” recombination, apparently mediated by the IRs. We also detected evidence for two distinct IR haplotypes and recombination between them in wild-caught P. estuarinus, as well as extensive inter-molecular recombination between SNPs in the LSC region. Together, these observations suggest that mitochondrial inheritance is not strictly maternal in these snails. The chloroplast-like architecture and repeat-mediated mitochondrial recombination we describe here raise fundamental questions regarding the origins and commonness of such architecture, whether and how recombination mediates mitochondrial genome evolution, and the role of genome architecture in driving cytoplasmic genome biology and the maintenance of cytoplasmic genomes.

A practice-based engineering degree, in which students worked on real-world, team-based projects from day one and throughout their course, was designed to address the need for employability skills in a rapidly changing world. Teaching mathematics in this way required a very different approach to the lecture-exam based model prevalent in most engineering degrees. In order to ensure all students developed the fundamental skills and knowledge required of engineers, a micro-credential based curriculum was developed where micro-credentials were mapped to projects and delivered “just-in-time”. The curriculum contained forty-eight explicit mathematical micro-credentials in the areas of measurement and geometry, algebra, calculus, and statistics and probability as well as many more micro-credentials in other areas of curriculum that contain mathematical skills, from physics to project budgeting. Mathematical competencies were used as a framework in the design and analysis of the micro-credentials. This paper presents a description of, and reflection on the successes and challenges in implementing this model of teaching mathematics in engineering.

Garlic (Allium sativum) has been historically used as a fly and pest control for horses, a practice that continues today. Recent research has shown garlic supplementation may cause Heinz body anemia in horses. The purpose of this study was to evaluate the incidence of Heinz body anemia in horses supplemented with incremental amounts of powdered garlic. The study included 12 horses divided into 4 groups (n=3), with horse serving as the experimental unit. Supplementation groups included: Control (0.0 g garlic/kg BW and 14.3 g molasses) and 3 supplementation rates: Low (0.0625 g garlic/kg BW and 14.3 g molasses), Medium (0.125 g garlic/kg BW and 14.3 g molasses), and High (0.1875 g garlic/kg BW and 14.3 g molasses) that were fed daily for 74 days. Horses were maintained on dormant tall fescue pasture and had ad libitum access to medium quality tall fescue hay throughout the study. Blood samples (jugular venipuncture) and weights were taken on day 0, 25, 50, and 74. Data were analyzed using Proc Mixed (SAS 9.4) for a completely randomized design, α = 0.05. Garlic supplementation reduced red blood cell count (P = 0.028) compared with Control and lymphocyte count (P = 0.046) showed a quadratic response to supplementation rate, with Low having the greatest lymphocyte count. A trend was noted for white blood cell count (P = 0.090) and hemoglobin count (P = 0.074) with garlic supplementation. Heinz body count (P = 0.406) was not affected by garlic supplementation. Results are interpreted to mean that garlic supplementation did not induce Heinz body anemia in horses; however, other blood constituents, including red blood cell and lymphocyte counts, were affected. Horses in the study were non-working, further research would be needed to evaluate any potential health impacts these variations in blood constituent concentrations would have in working horses.

In 1952, Gilbert S. Daniels published his seminal report “The ’Average Man’?” which examined the measurements of 4,063 active United States Air Force personnel (Daniels. 1952). This report detailed a profound yet simple finding: that after eliminating for ten common anthropometric measurements, no one person meets the average for all body dimensions. We analyzed Two anthropometric data bases over two studies, following an updated version of Daniels’ (1952) original method. The biggest changes were not eliminating the top and bottom percentiles of the population, and adjusted the calculation to find middle 25-30%. The databases examined were ANSUR II (Gorden et al., 2014) and CAESAR (Robinette et al., 2002), to see how civilian and military populations compare. The results of both studies were generally consistent with Daniels (1952), however we did discover that some individuals were able to meet the criteria for average after ten measurements. The best performance took place in the ANSUR II combined condition, all three individuals were men. This last part is especially important to note, as the combined sample eliminated women out of the sample faster than men. Being eliminated after the sixth measurement. This confirms a potential bias to combining men’s and women’s measurements without great care. Based on our analysis of modern anthropometric databases using Daniels’ original method it is clear that, while we found some “average people”, the significance of its findings holds true. This is not to say we should ignore the average, but we should understand its use in context and strive to go beyond it. Thinking past a formative understanding of how people are shaped, and instead into what is needed to create well-fitting products for a specific population. Examples spanning several industries merely scratch the surface of what needs to be addressed. Looking around as we go through our day, minor and major inconveniences become apparent. They cannot be all fixed at once, but through diligent research and thoughtful design we can use the principles of universal design to our advantage. Looking ahead to the next 70 years, a continued growth in optimizing products for the individual user and helping these users understand why these optimizations matter is not just desirable, but important.

Problem-based learning (PBL) has a history of producing strong educational results in engineering; however, global society is challenged by highly complex environmental, socio-political and technic...

This article presents a case description of the design decisions, delivery methods, and assessment framework for an entrepreneurship micro-credential in a newly developed practice-based engineering degree, which was codesigned with industry partners who called for the integration of innovation, proactivity, and creativity (i.e., characteristics of entrepreneurship) into engineering education. Students undertake the micro-credential via online and face-to-face modules. Assessment is competency-based, requiring students to apply the theoretical knowledge provided in the micro-credential to an industry-based project. By participating in the micro-credential, it is expected that students will have a better understanding of how products and services can address customer needs, and how opportunities for product and service enhancement can create opportunities for growing the offerings of engineering. Several insights into the strengths, limitations, and design considerations for entrepreneurship micro-credentials in an engineering education context are outlined in addition to suggestions for improvement.

Most eukaryotic microbial diversity is uncultivated, under-studied and lacks nuclear genome data. Mitochondrial genome sampling is more comprehensive, but many phylogenetically important groups remain unsampled. Here, using a single-cell sorting approach combining tubulin-specific labelling with photopigment exclusion, we sorted flagellated heterotrophic unicellular eukaryotes from Pacific Ocean samples. We recovered 206 single amplified genomes, predominantly from underrepresented branches on the tree of life. Seventy single amplified genomes contained unique mitochondrial contigs, including 21 complete or near-complete mitochondrial genomes from formerly under-sampled phylogenetic branches, including telonemids, katablepharids, cercozoans and marine stramenopiles, effectively doubling the number of available samples of heterotrophic flagellate mitochondrial genomes. Collectively, these data identify a dynamic history of mitochondrial genome evolution including intron gain and loss, extensive patterns of genetic code variation and complex patterns of gene loss. Surprisingly, we found that stramenopile mitochondrial content is highly plastic, resembling patterns of variation previously observed only in plants. Single-cell sequencing of flagellated heterotrophic unicellular eukaryotes provides insights into the dynamics of mitochondrial genome evolution.

Baker's yeast has a finite lifespan and ages in two ways: a mother cell can only divide so many times (its replicative lifespan), and a non-dividing cell can only live so long (its chronological lifespan). Wild and laboratory yeast strains exhibit natural variation for each type of lifespan, and the genetic basis for this variation has been generalized to other eukaryotes, including metazoans. To date, yeast chronological lifespan has chiefly been studied in relation to the rate and mode of functional decline among non-dividing cells in nutrient-depleted batch culture. However, this culture method does not accurately capture two major classes of long-lived metazoan cells: cells that are terminally differentiated and metabolically active for periods that approximate animal lifespan (e.g. cardiac myocytes), and cells that are pluripotent and metabolically quiescent (e.g. stem cells). Here, we consider alternative ways of cultivating Saccharomyces cerevisiae so that these different metabolic states can be explored in non-dividing cells: (i) yeast cultured as giant colonies on semi-solid agar, (ii) yeast cultured in retentostats and provided sufficient nutrients to meet minimal energy requirements, and (iii) yeast encapsulated in a semisolid matrix and fed ad libitum in bioreactors. We review the physiology of yeast cultured under each of these conditions, and explore their potential to provide unique insights into determinants of chronological lifespan in the cells of higher eukaryotes.

Eugene R Viscusi,1 Franck Skobieranda,2 David G Soergel,2 Emily Cook,2 David A Burt,2 Neil Singla3 1Department of Anesthesiology, Sidney Kimmel Medical College at Thomas Jefferson University, Philadelphia, PA, USA; 2Trevena Inc., Chesterbrook, PA, USA; 3Lotus Clinical Research, LLC, Pasadena, CA, USA Purpose: Oliceridine is a novel G protein-biased µ-opioid receptor agonist designed to provide intravenous (IV) analgesia with a lower risk of opioid-related adverse events (ORAEs) than conventional opioids. Patients and methods: APOLLO-1 (NCT02815709) was a phase III, double-blind, randomized trial in patients with moderate-to-severe pain following bunionectomy. Patients received a loading dose of either placebo, oliceridine (1.5mg), or morphine (4mg), followed by demand doses via patient-controlled analgesia (0.1, 0.35, or 0.5mg oliceridine, 1mg morphine, or placebo). The primary endpoint compared the proportion of treatment responders through 48 hours for oliceridine regimens and placebo. Secondary outcomes included a composite measure of respiratory safety burden (RSB, representing the cumulative duration of respiratory safety events) and the proportion of treatment responders vs morphine. Results: Effective analgesia was observed for all oliceridine regimens, with responder rates of 50%, 62%, and 65.8% in the 0.1 mg, 0.35 mg, and 0.5 mg regimens, respectively(allP

Long-term preservation of laboratory strains of Chlamydomonas reinhardtii has historically involved either liquid nitrogen cryopreservation, which is expensive and labor intensive, or storage on agar plates, which requires frequent transfer to new plates, and which may leave samples susceptible to contamination as well as genetic drift and/or selection. The emergence of C. reinhardtii as a model organism for genetic analysis and experimental evolution has produced an increasing demand for an efficient method to cryopreserve C. reinhardtii populations. The GeneArt™ Cryopreservation Kit for Algae provides the first method for algal storage at −80°C; however, little is known about how this method affects recovery of different clones, much less polyclonal populations. Here, we compare postfreeze viability of clonal and genetically mixed samples frozen at −80°C using GeneArt™ or cryopreserved using liquid nitrogen. We find that the GeneArt™ protocol yields similar percent recoveries for some but not all clonal cultures, when compared to archiving via liquid N2. We also find that relative frequency of different strains recovered from genetically mixed populations can be significantly altered by cryopreservation. Thus, while cryopreservation using GeneArt™ is an effective means for archiving certain clonal populations, it is not universally so. Strain-specific differences in freeze–thaw tolerance complicate the storage of different clones, and may also bias the recovery of different genotypes from polyclonal populations.

Total ankle arthroplasty (TAA) is a viable treatment option for end-stage ankle arthritis. However, implant survivorship remains an important consideration. Concerns regarding early component loosening with the low-profile tibial tray utilized by fourth-generation TAA systems have been raised in the literature. We have previously described our preliminary outcomes of a hybrid technique combining a stemmed intramedullary tibial component with a chamfer-cut talar component for TAA. A retrospective study comparing short-term outcomes of the tibial component between a standard fourth-generation TAA system versus our hybrid technique was performed. 46 patients with a minimum of 1-year follow up were included in the analyses. There were 25 subjects in the standard implant cohort utilizing a low-profile tibial tray, and 21 subjects in the hybrid group utilizing a stemmed intramedullary tibial component. No statistically significant difference between the demographics of each group was found. The rate of tibial component subsidence was 8% (n = 2) in the standard implant group, and 0% (n = 0) in the hybrid group, though this did not meet statistical significance (p = .49). Mean time to subsidence was 6 months, and revision rate due to tibial component subsidence was 2.1% (n = 1). Periprosthetic lucency was present on most recent follow-up radiographs in 32% and 9.5% of ankles in the standard and hybrid groups, respectively (p = .08). Despite prior concerns for tibial component subsidence with the standard fourth-generation system, we demonstrated low rates in both implant groups. Additional studies are needed to further explore factors that may predispose patients to early tibial component subsidence and resulting implant failure.

The productivity of a biological community often correlates with its diversity. In the microbial world this phenomenon can sometimes be explained by positive, density-dependent interactions such as cross-feeding and syntrophy. These metabolic interactions help account for the astonishing variety of microbial life and drive many of the biogeochemical cycles without which life as we know it could not exist. While it is difficult to recapitulate experimentally how these interactions evolved among multiple taxa, we can explore in the laboratory how they arise within one. These experiments provide insight into how different bacterial ecotypes evolve and from these, possibly new “species.” We have previously shown that in a simple, constant environment a single clone of Escherichia coli can give rise to a consortium of genetically and phenotypically differentiated strains, in effect, a set of ecotypes, that coexist by cross-feeding. We marked these different ecotypes and their shared ancestor by integrating fluorescent protein into their genomes and then used flow cytometry to show that each evolved strain is more fit than the shared ancestor, that pairs of evolved strains are fitter still, and that the entire consortium is the fittest of all. We further demonstrate that the rank order of fitness values agrees with estimates of yield, indicating that an experimentally evolved consortium more efficiently converts primary and secondary resources to offspring than its ancestor or any member acting in isolation. IMPORTANCE Polymicrobial consortia occur in both environmental and clinical settings. In many cases, diversity and productivity correlate in these consortia, especially when sustained by positive, density-dependent interactions. However, the evolutionary history of such entities is typically obscure, making it difficult to establish the relative fitness of consortium partners and to use those data to illuminate the diversity-productivity relationship. Here, we dissect an Escherichia coli consortium that evolved under continuous glucose limitation in the laboratory from a single common ancestor. We show that a partnership consisting of cross-feeding ecotypes is better able to secure primary and secondary resources and to convert those resources to offspring than the ancestral clone. Such interactions may be a prelude to a special form of syntrophy and are likely determinants of microbial community structure in nature, including those having clinical significance such as chronic infections.

Interprofessional collaboration is fundamental to providing optimal patient care. The readiness of the team entering a framework of interprofessional collaborative practice is critical to its success. In this study, we conducted an interprofessional education (IPE) activity for medical and nursing students in an acute care setting. Over nine occasions, 21 student pairs (one nursing and one medical student per pair) jointly assessed a patient and created a list of problems and interventions to achieve the patient’s goals. Immediately after the activity, students were debriefed to gain insight into their experiences. Debriefing sessions were audiotaped and analysed using a phenomenological approach and four major themes were identified. Overall, students felt responsible for representing their profession and were initially apprehensive about the interprofessional task. Nevertheless, they identified their own shortcomings and recognized the value in their partner’s approach. These realizations promoted convergence on a shared vision to provide optimal care for patients as a team. Acknowledging and understanding these perceptions may help design better ways to improve patient care. This educational model may be utilized by others who are seeking IPE activities in acute care.

Global reward management plays a fundamental role in supporting the attraction, motivation and retention of employees, and yet recent research has underscored limitations of the dominant balance sheet approach, including inequity between host country national and expatriate staff. To shed light on how reward in international contexts can be structured to address issues of fairness and equity, this study explores approaches to global reward in international non-governmental organizations (INGOs), an underexplored context where fairness may be particularly salient. Through an inductive study of 15 INGOs, we show how organizations are reconceptualizing global reward systems by questioning dominant assumptions of the expatriate workforce and the jobs they do, and broadening consideration of reward to include both monetary and non-monetary components. Doing so enables incremental shifts toward strategic alignment of global reward with underlying social values. Our findings provide important insights for organizations operating internationally about how global reward can be structured to address concerns of fairness, while still enabling organizations to meet their demands for particular skills.

Highlights • Cortical GABA levels correlate with contrast detection thresholds. • GABA appears to reduce contrast response gain and intrinsic noise. • Changes in contrast discrimination can be characterised by the Wilson-Cowan model., High levels of GABA (gamma-aminobutyric acid, the brain's primary inhibitory neurotransmitter) are associated with enhanced cognitive and perceptual performance. It has been proposed that these effects result from GABA reducing neural noise or variability, but the precise mechanisms remain unknown. We have measured how individual differences in GABA concentration in the visual cortex are related to performance on a visual contrast discrimination task. Our results reveal that the facilitatory strength of the typical “dipper” function elicited by this task is strongly correlated with GABA concentration. A simple, biologically plausible, network model comprising excitatory and suppressive neural populations accounts for the data well and indicates that the strength of suppression increases as GABA concentration increases. Inter-individual variations in GABA were correlated both with the inhibition strength of the model (mimicking the effect of GABA) and, inversely, with the magnitude of the response criterion. This enhanced suppression has the dual effect of suppressing noise and reducing the gain of the neural response. Our findings thus suggest that the changes in performance conferred by high GABA concentration are mediated by both a reduction of noise and, paradoxically, a reduction in neural, but not perceptual, sensitivity.

Community productivity often correlates with diversity. In the microbial world this phenomenon can sometimes be explained by highly-specific metabolic interactions that include cross-feeding and syntrophy. Such interactions help account for the astonishing variety of microbial life, and drive many of the biogeochemical cycles without which life as we know it could not exist. While it is difficult to recapitulate experimentally how these interactions evolved among multiple taxa, we can explore in the laboratory how they arise within one. These experiments provide insight into how different bacterial ecotypes evolve and from these, possibly new ‘species.’ We have previously shown that in a simple, constant environment a single clone ofE. colican give rise to a consortium of genetically-and physiologically-differentiated strains, in effect, a set of ecotypes, that coexist by cross-feeding. We marked these different ecotypes and their shared ancestor by integrating fluorescent protein into their genomes. We then used flow cytometry to show that each strain by itself is more fit than the shared ancestor, that pairs of evolved strains are fitter still, and that the entire consortium is fittest of all. We further demonstrate that the rank order of fitness values agrees with estimates of yield, indicating that an experimentally evolved consortium more efficiently converts resources to offspring than its ancestor or any member acting in isolation.ImportanceIn the microbial world, diversity and productivity of communities and consortia often correlate positively. However, it is challenging to tease apart a consortium whose members have co-evolved, and connect estimates of their fitness and the fitness of their ancestor(s) with estimates of productivity. Such analyses are prerequisite to understanding the evolutionary origins of all biological communities. Here we dissect anE. coliconsortium that evolved in the laboratory and show that cooperative interactions are favored under continuous glucose limitation because a partnership of ecotypes is better able to scavenge all available resources and more efficiently convert those resources to offspring than any single individual. Such interactions may be a prelude to a special form of syntrophy, and are likely to be key determinants of microbial community structure in nature, including those having clinical significance, such as chronic infections.

Significance Horizontal gene transfer (HGT) is the transfer of genetic information between genomes by a route other than from parent to offspring. Of particular interest here are the transfers of transporter-encoding genes, which can allow an organism to utilize a new metabolite, often via the acquisition of a single foreign gene. Here we have identified a range of HGT events of transporter-encoding genes, characterized the substrate preferences for each HGT encoded protein, and demonstrated that the gain of one of these HGTs can provide yeast with a distinct competitive advantage in a given environment. This has wide implications for understanding how acquisition of single genes by HGT can drastically influence the environments fungi can colonize., Many microbes acquire metabolites in a “feeding” process where complex polymers are broken down in the environment to their subunits. The subsequent uptake of soluble metabolites by a cell, sometimes called osmotrophy, is facilitated by transporter proteins. As such, the diversification of osmotrophic microorganisms is closely tied to the diversification of transporter functions. Horizontal gene transfer (HGT) has been suggested to produce genetic variation that can lead to adaptation, allowing lineages to acquire traits and expand niche ranges. Transporter genes often encode single-gene phenotypes and tend to have low protein–protein interaction complexity and, as such, are potential candidates for HGT. Here we test the idea that HGT has underpinned the expansion of metabolic potential and substrate utilization via transfer of transporter-encoding genes. Using phylogenomics, we identify seven cases of transporter-gene HGT between fungal phyla, and investigate compatibility, localization, function, and fitness consequences when these genes are expressed in Saccharomyces cerevisiae. Using this approach, we demonstrate that the transporters identified can alter how fungi utilize a range of metabolites, including peptides, polyols, and sugars. We then show, for one model gene, that transporter gene acquisition by HGT can significantly alter the fitness landscape of S. cerevisiae. We therefore provide evidence that transporter HGT occurs between fungi, alters how fungi can acquire metabolites, and can drive gain in fitness. We propose a “transporter-gene acquisition ratchet,” where transporter repertoires are continually augmented by duplication, HGT, and differential loss, collectively acting to overwrite, fine-tune, and diversify the complement of transporters present in a genome.

Highlights • Cortical GABA levels correlate with visuo-spatial IQ and surround suppression. • Higher levels predict higher intelligence and stronger surround suppression. • Results provide mechanism linking surround suppression and intelligence. • Results suggest role of cortical GABA levels in determining cognitive performance., Early psychological researchers proposed a link between intelligence and low-level perceptual performance. It was recently suggested that this link is driven by individual variations in the ability to suppress irrelevant information, evidenced by the observation of strong correlations between perceptual surround suppression and cognitive performance. However, the neural mechanisms underlying such a link remain unclear. A candidate mechanism is neural inhibition by gamma-aminobutyric acid (GABA), but direct experimental support for GABA-mediated inhibition underlying suppression is inconsistent. Here we report evidence consistent with a global suppressive mechanism involving GABA underlying the link between sensory performance and intelligence. We measured visual cortical GABA concentration, visuo-spatial intelligence and visual surround suppression in a group of healthy adults. Levels of GABA were strongly predictive of both intelligence and surround suppression, with higher levels of intelligence associated with higher levels of GABA and stronger surround suppression. These results indicate that GABA-mediated neural inhibition may be a key factor determining cognitive performance and suggests a physiological mechanism linking surround suppression and intelligence.

This arts- based project creatively introduces residents to photography, self-portraiture and narratives to document the longitudinal journey of becoming a family physician. Visual arts and writing can foster reflection: an important skill to cultivate in developing physicians. Unfortunately, arts based programs are lacking in many residency programs. Tools and venues that nourish physician well being and resilience may be important in today’s changing healthcare environment and epidemic of physician burnout. Residents created self-portraits with accompanying narratives throughout their three-year training. Analysis of the portraits and accompanying narratives completed the assessment. Residents created a body of work that includes 182 creative and deeply personal portraits and narratives. The five most frequent themes of portraits included “Residency is Difficult,” “Hobbies,” “Family,” “Growing as a Doctor,” and “Coping Mechanisms.” Self-portrait photography and reflection gives insight into the journey of becoming a family medicine physician at a deeply personal and professional level. Further partnerships between residency programs and the arts should be explored to promote reflection.

Diabetic foot ulcers (DFU) are a major health problem associated with diabetes mellitus. Impaired nitric oxide (NO) production has been shown to be a major contributor to the dysregulation of healing in DFU. The level of impairment is not known primarily due to challenges with measuring NO. Herein, we report the actual level of NO produced by human dermal fibroblasts cultured under normal and high glucose conditions. Fibroblasts produce the extracellular matrix, which facilitate the migration of keratinocytes to close wounds. The results show that NO production was significantly higher in normal glucose compared to high glucose conditions. The real-time NO detected was compared to the nitrite present in the culture media and there was a direct correlation between real-time NO and nitrite in normal glucose conditions. However, real-time NO detection and nitrite measurement did not correlate under high glucose conditions. The inducible nitric oxide synthase (iNOS) enzyme responsible for NO production was upregulated in normal and high glucose conditions and the proliferation rate of fibroblasts was not statistically different in all the treatment groups. Relying only on nitrite to assess NO production is not an accurate determinant of the NO present in the wound bed in pathological states such as diabetes mellitus.

Genome analysis using pulsed-field gel electrophoresis (PFGE) has been used in applications ranging from typing bacterial strains to radiobiology to cancer research. While methods for running PFGE have been significantly improved since its invention, the method for preparing chromosomal DNA itself has remained essentially unchanged. This limits the applicability of PFGE, especially when analyses require many samples. We have streamlined sample preparation for routine applications of PFGE through the use of deep-well 48-well plates. Besides saving time, our protocol has the added advantage of reducing the volume of expensive reagents. Our improved protocol enables us to reduce throughput time and simplify the procedure, facilitating wider application of PFGE-based analyses in the laboratory.

This article discusses visual literacy, its connection to information literacy, and its significance to scientific disciplines. It includes a case study from Washington and Lee (W&L) University that showcases how libraries can integrate visual literacy instruction into STEM courses. In the study, two W&L Library staff members partnered with one W&L visiting assistant professor of physics to transform a common assignment, the academic poster, into a digital form of visual communication. This shift resulted in a revised evaluative rubric and led to enhanced library led instruction focusing on information literacy, visual literacy, and digital literacy skills.

Microbes rarely exist in isolation, rather, they form intricate multi-species communities that colonize our bodies and inserted medical devices. However, the efficacy of antimicrobials is measured in clinical laboratories exclusively using microbial monocultures. Here, to determine how multi-species interactions mediate selection for resistance during antibiotic treatment, particularly following drug withdrawal, we study a laboratory community consisting of two microbial pathogens. Single-species dose responses are a poor predictor of community dynamics during treatment so, to better understand those dynamics, we introduce the concept of a dose-response mosaic, a multi-dimensional map that indicates how species’ abundance is affected by changes in abiotic conditions. We study the dose-response mosaic of a two-species community with a ‘Gene × Gene × Environment × Environment’ ecological interaction whereby Candida glabrata, which is resistant to the antifungal drug fluconazole, competes for survival with Candida albicans, which is susceptible to fluconazole. The mosaic comprises several zones that delineate abiotic conditions where each species dominates. Zones are separated by loci of bifurcations and tipping points that identify what environmental changes can trigger the loss of either species. Observations of the laboratory communities corroborated theory, showing that changes in both antibiotic concentration and nutrient availability can push populations beyond tipping points, thus creating irreversible shifts in community composition from drug-sensitive to drug-resistant species. This has an important consequence: resistant species can increase in frequency even if an antibiotic is withdrawn because, unwittingly, a tipping point was passed during treatment. Single-species antibiotic dose response is a poor predictor of multi-species community dynamics because it cannot foresee the tipping points that cause irreversible changes in resistance that persist even when treatment stops.

This Excessive sugar-sweetened beverage (SSB) consumption and low health literacy skills have emerged as two public health concerns in the United States (US); however, there is limited research on how to effectively address these issues among adults. As guided by health literacy concepts and the Theory of Planned Behavior (TPB), this randomized controlled pilot trial applied the RE-AIM framework and a mixed methods approach to examine a sugar-sweetened beverage (SSB) intervention (SipSmartER), as compared to a matched-contact control intervention targeting physical activity (Move-More). Both 5-week interventions included two interactive group sessions and three support telephone calls. Executing a patient-centered developmental process, the primary aim of this paper was to evaluate patient feedback on intervention content and structure. The secondary aim was to understand the potential reach (i.e., proportion enrolled, representativeness) and effectiveness (i.e. health behaviors, theorized mediating variables, quality of life) of SipS- martER. Twenty-five participants were randomized to SipSmartER (n = 14) or MoveMore (n = 11). Participants’ intervention feedback was positive, ranging from 4.2 - 5.0 on a 5-point scale. Qualitative assessments reavealed several opportuneties to improve clarity of learning materials, enhance instructions and communication, and refine research protocols. Although SSB consumption decreased more among the SipS-martER participants (?256.9 ± 622.6 kcals), there were no significant group differences when compared to control participants (?199.7 ± 404.6 kcals). Across both groups, there were significant improvements for SSB attitudes, SSB behavioral intentions, and two media literacy constructs. The value of using a patient-centered approach in the developmental phases of this intervention was apparent, and pilot findings suggest decreased SSB may be achieved through targeted health literacy and TPB strategies. Future efforts are needed to examine the potential public health impact of a large-scale trial to address health literacy and reduce SSB.

Bovine herpesvirus 1 (BHV-1), an alphaherpesvirinae subfamily member, establishes latency in sensory neurons. Elevated corticosteroid levels, due to stress, reproducibly triggers reactivation from latency in the field. A single intravenous injection of the synthetic corticosteroid dexamethasone (DEX) to latently infected calves consistently induces reactivation from latency. Lytic cycle viral gene expression is detected in sensory neurons within 6 h after DEX treatment of latently infected calves. These observations suggested that DEX stimulated expression of cellular genes leads to lytic cycle viral gene expression and productive infection. In this study, a commercially available assay—Bovine Gene Chip—was used to compare cellular gene expression in the trigeminal ganglia (TG) of calves latently infected with BHV-1 versus DEX-treated animals. Relative to TG prepared from latently infected calves, 11 cellular genes were induced more than 10-fold 3 h after DEX treatment. Pentraxin three, a regulator of innate immunity and neurodegeneration, was stimulated 35- to 63-fold after 3 or 6 h of DEX treatment. Two transcription factors, promyelocytic leukemia zinc finger (PLZF) and Slug were induced more than 15-fold 3 h after DEX treatment. PLZF or Slug stimulated productive infection 20- or 5-fold, respectively, and Slug stimulated the late glycoprotein C promoter more than 10-fold. Additional DEX-induced transcription factors also stimulated productive infection and certain viral promoters. These studies suggest that DEX-inducible cellular transcription factors and/or signaling pathways stimulate lytic cycle viral gene expression, which subsequently leads to successful reactivation from latency in a small subset of latently infected neurons.

In the version of this Article originally published, the following sentence was missing from the Acknowledgements: “R.E.B. is an EPSRC Healthcare Technologies Impact Fellow EP/N033671/1; I.G. is funded by ERC Consolidator grant 647292 MathModExp; A.J.P.B., N.A.R.G. and A.T. were funded by BBSRC grant BB/F00513X/1; K.H., I.G., S.N. and E.C. were funded by BBSRC grant BB/F005210/2.” This text has now been added.

Preliminary studies have shown the effectiveness of multivariate analysis (MVA) for drug identification from energy-dispersive X-ray diffraction patterns. A statistical model to predict drug content from the diffraction profile of a sample of mixed composition was developed by applying MVA to both experimental and simulated data. Separate data-sets were used for building and testing the models. Both experimental and simulated data were used and the MVA predictions compared. Experimental data included diffraction patterns from small (5 mm diameter) drug samples with various cutting agents, acquired with a HPGe detector; simulated data included diffraction patterns of samples including materials simulating drugs (i.e., materials featuring sharp diffraction peaks in the relevant momentum transfer range) and typical packaging materials. Both a HPGe detector (energy resolution 0.7 keV at 59.5 keV) and a CZT detector (energy resolution 4 keV at all energies) were simulated. MVA was used to predict the drug content. In all cases different statistics were applied to assess the detection limits of the models. Multivariate analysis has proved effective in both identifying the presence of a drug and its concentration. Due to the large contribution to peak broadening given by angular resolution, no significant decrease in accuracy has been found when using CZT with respect to HPGe data.

Biofilm formation (BF) in the setting of candiduria has not been well studied. We determined BF and MIC to antifungals in Candida spp. isolates grown from urine samples of patients and performed a retrospective chart review to examine the correlation with risk factors. A total of 67 Candida spp. isolates were grown from urine samples from 55 patients. The species distribution was C. albicans (54%), C. glabrata (36%), and C. tropicalis (10%). BF varied greatly among individual Candida isolates but was stable in sequential isolates during chronic infection. BF also depended on the growth medium and especially in C. albicans was significantly enhanced in artificial urine (AU) compared to RPMI medium. In nine of the C. albicans strains BF was 4- to 10-fold higher in AU, whereas in three of the C. albicans strains and two of the C. glabrata strains higher BF was measured in RPMI medium than in AU. Determination of the MICs showed that planktonic cells of all strains were susceptible to amphotericin B (AMB) and caspofungin (CASPO) and that three of the C. glabrata strains and two of the C. albicans strains were resistant to fluconazole (FLU). In contrast, all biofilm-associated adherent cells were resistant to CASPO and FLU. The biofilms of 14 strains (28%) were sensitive to AMB (MIC 50 of Candida strains that varies greatly among clinical strains and is dependent on the growth medium. Resistance to AMB is associated with higher BF in AU, which may represent the more physiologic medium to test BF. Future studies should address whether in vitro BF can predict treatment failure in vivo.