Stakeholder Recommendations to Refine the Fitness-to-Drive Screening Measure

In developing the web-based Fitness-to-Drive Screening Measure (FTDS) and keyform (results output) for use to identify at-risk older drivers, we examined the needs, perspectives, and suggestions of three stakeholders groups: occupational therapy practitioners, certified driver rehabilitation specialists (CDRSs), and family members/caregivers. We conducted three focus groups, which were moderated, recorded, transcribed, and analyzed using directed content analysis. Respondents in two focus groups also rated FTDS aspects (e.g., ease of use, format, and relevance), using a visual analog scale (VAS, 0-10 scale with 10 being excellent). All three stakeholder groups contributed to the development of the web-based FTDS. Results from occupational therapy practitioners addressed face validity, appearance, wording, and usability; CDRSs informed follow-up recommendations; and family members/caregivers provided keyform feedback. High VAS ratings (\u3e 7 on 1-10 scale) from the CDRSs (8.4, SD+0.8) and family members/caregivers (9.01, SD+1.02) indicated FTDS acceptability. Overall, our findings support the measure’s utility and acceptability among these users. As such, the FTDS may position family members/caregivers to identify at-risk older drivers, facilitate targeted discussions of driving difficulty among occupational therapists and their clients, and afford OT-CDRS an entry point for intervention and clinical decision making

An Integrative Review on Teen Distracted Driving for Model Program Development

Distracted driving, especially driver inattention, is associated with high levels of crash-related fatalities and injury. Teen novice drivers are one of the groups most likely to drive distracted and to suffer its consequences. Teens have a higher risk of engaging in texting or secondary tasks, e.g., eating while driving. Distracted driving interventions to date aim to improve teen and societal safety, but few have achieved effectiveness. A need exists for effective evidence-based distracted driving interventions. We used an integrative review to identify rigorous evidence, and inform the development of a teen distracted driving educational intervention. This five-step review included: identifying the research problem; collecting literature; evaluating literature; synthesizing data; and presenting results. We searched 6 databases, identifying 185 articles. Following three rounds of inclusion screening (title, abstract, and full-text), captured according to a PRISMA flow chart, 17 studies met inclusion. We categorized these studies, conducted in the U.S., as five intervention types that used approaches including presentations, videos or instructional programs, education or training programs, driving simulator training, in-vehicle monitoring or feedback, and integrated programs. Study designs included randomized controlled trials pre-post, quasi-experimental, and experimental designs with prospective longitudinal cohorts. The studies were heterogeneous in design, intervention and outcome. However, three core themes emerged across studies: i.e., hazard awareness, hazard mitigation and attention maintenance are primary critically necessary skills to prevent distracted driving; engaging a parent or adult as a partner in the intervention process from classroom to car contributed to the effectiveness of the intervention; and leveraging technology in training enhanced the effectiveness of the intervention. Study limitations pertained to a focus on short-term effects; sampling distributions that did not account for gender, age, race, and/or ethnicity; types of interventions; and bias. The limitations affect the generalizability of included study findings and, potentially, the review findings, as they may not apply to populations or contexts outside those synopsized. Strengths included our team's expertise in conducting evidence-based reviews, support of a health science librarian, and use of international review guidelines. As an outcome, we are applying findings of the integrated review to develop a computer-based training addressing teen distracted driving

Leishmania amazonensis Arginase Compartmentalization in the Glycosome Is Important for Parasite Infectivity

Abstract In Leishmania, de novo polyamine synthesis is initiated by the cleavage of L-arginine to urea and L-ornithine by the action of arginase (ARG, E.C. 3.5.3.1). Previous studies in L. major and L. mexicana showed that ARG is essential for in vitro growth in the absence of polyamines and needed for full infectivity in animal infections. The ARG protein is normally found within the parasite glycosome, and here we examined whether this localization is required for survival and infectivity. First, the localization of L. amazonensis ARG in the glycosome was confirmed in both the promastigote and amastigote stages. As in other species, arg 2 L. amazonensis required putrescine for growth and presented an attenuated infectivity. Restoration of a wild type ARG to the arg 2 mutant restored ARG expression, growth and infectivity. In contrast, restoration of a cytosoltargeted ARG lacking the glycosomal SKL targeting sequence (argDSKL) restored growth but failed to restore infectivity. Further study showed that the ARGDSKL protein was found in the cytosol as expected, but at very low levels. Our results indicate that the proper compartmentalization of L. amazonensis arginase in the glycosome is important for enzyme activity and optimal infectivity. Our conjecture is that parasite arginase participates in a complex equilibrium that defines the fate of L-arginine and that its proper subcellular location may be essential for this physiological orchestration

Histamine selectively interrupts VE-cadherin adhesion independently of capacitive calcium entry

Histamine is an important agent of innate immunity, transiently increasing the flux of immune-competent molecules from the vascular space to the tissues and then allowing rapid restoration of the integrity of the endothelial barrier. In previous work we found that histamine alters the endothelial barrier by disrupting cell-cell adhesion and identified VE-cadherin as an essential participant in this process. The previous work did not determine whether histamine directly interrupted VE-cadherin adhesion, whether the effects of histamine were selective for cadherin adhesion, or whether capacitive calcium flux across the cell membrane was necessary for the effects of histamine on cell-cell adhesion. In the current work we found that histamine directly interrupts adhesion of L cells expressing the type 1 histamine (H1) receptor and VE-cadherin to a VE-cadherin-Fc fusion protein. In contrast, integrin-mediated adhesion to fibronectin of the same L cells expressing the H1 receptor was not affected by histamine, demonstrating that the effects of histamine are selective for cadherin adhesion. Some of the effects of many edemagenic agonists on endothelium are dependent on the capacitive flux of calcium across the endothelial cell membrane. Blocking capacitive calcium flux with LaCl3 did not prevent histamine from interrupting VE-cadherin adhesion of transfected L cells, nor did it prevent histamine from interrupting cell-cell adhesion of human umbilical vein endothelial cells. These data support the contentions that histamine directly and selectively interrupts cadherin adhesion and this effect on cadherin adhesion is independent of capacitive calcium flux. endothelium; integrin; human histamine receptor H1; vascular endothelial cadherin ACUTE INFLAMMATORY EDEMA is an important component of innate immunity that facilitates the transfer of immune-competent molecules from the vascular space to the tissues. Histamine and serotonin are well-acknowledged physiological agonists of acute inflammatory edema that disrupt cell-cell apposition in postcapillary venules Vascular endothelial (VE)-cadherin is an endothelial unique cadherin that mediates cell-cell adhesion of endothelial cells (17). Function-interrupting antibody to VE-cadherin increases permeability of endothelium in vivo, indicating that integrity of VE-cadherin adhesion is essential to the integrity of the endothelial barrier (9, 16). It was of note that although antibody to VE-cadherin increased in vivo endothelial permeability within 1 h of its administration, no light microscopic changes were evident in the endothelium even at 2 h, demonstrating that no large gaps had formed between the cells. However, similar to the effects of histamine and serotonin, transmission electron micrographs detected very small gaps in lung microvessels exposed to antibody to VE-cadherin (9). ECV304 cells are a transformed bladder epithelial cell line that express the type 1 histamine receptor (H1) and P-and N-cadherin, but not VE-cadherin. When mock-transfected ECV304 cells were exposed to histamine, there was no change in the electrical resistance of the cell-cell barrier they created (24). However, when ECV304 cells transfected with VEcadherin were exposed to histamine, the electrical resistance of the cell-cell barrier fell, identifying an important role for VE-cadherin in the response to histamine of a monolayer of polarized cells (24). ECV304 cells express claudin 2 and develop tight junctions (5). This tight junction is a necessary component of the electrical resistance decreased by histamine. The observation that histamine decreased the integrity of the cell-cell barrier created by VE-cadherin transfected ECV304 cells did not discriminate between effects of histamine on an interaction of VE-cadherin with the tight junction versus a direct effect of histamine on VE-cadherin adhesion. Histamine and other agonists that increase endothelial permeability initiate signaling in endothelial cells that increases cell calcium (6, 7). Although some of the increase in cell calcium caused by these agonists reflects release of calcium from intracellular stores, capacitive flux of calcium across the cell membrane is also activated, and the capacitive calcium flux is necessary for some of the effects of these agonists on endothelial cells In these investigations we used a model of cadherin adhesion that lacks tight junctions and asked whether histamine directly affects VE-cadherin adhesion as opposed to affecting an interaction between VE-cadherin and the tight junction. Our results indicate that histamine directly affects VE-cadherin adhesion. Using a similar model of integrin-based adhesion, we foun

The sialic acid binding activity of the S protein facilitates infection by porcine transmissible gastroenteritis coronavirus

<p>Abstract</p> <p>Background</p> <p>Transmissible gastroenteritis virus (TGEV) has a sialic acid binding activity that is believed to be important for enteropathogenicity, but that has so far appeared to be dispensable for infection of cultured cells. The aims of this study were to determine the effect of sialic acid binding for the infection of cultured cells under unfavorable conditions, and comparison of TGEV strains and mutants, as well as the avian coronavirus IBV concerning their dependence on the sialic acid binding activity.</p> <p>Methods</p> <p>The infectivity of different viruses was analyzed by a plaque assay after adsorption times of 5, 20, and 60 min. Prior to infection, cultured cells were either treated with neuraminidase to deplete sialic acids from the cell surface, or mock-treated. In a second approach, pre-treatment of the virus with porcine intestinal mucin was performed, followed by the plaque assay after a 5 min adsorption time. A student's t-test was used to verify the significance of the results.</p> <p>Results</p> <p>Desialylation of cells only had a minor effect on the infection by TGEV strain Purdue 46 when an adsorption period of 60 min was allowed for initiation of infection. However, when the adsorption time was reduced to 5 min the infectivity on desialylated cells decreased by more than 60%. A TGEV PUR46 mutant (HAD3) deficient in sialic acid binding showed a 77% lower titer than the parental virus after a 5 min adsorption time. After an adsorption time of 60 min the titer of HAD3 was 58% lower than that of TGEV PUR46. Another TGEV strain, TGEV Miller, and IBV Beaudette showed a reduction in infectivity after neuraminidase treatment of the cultured cells irrespective of the virion adsorption time.</p> <p>Conclusions</p> <p>Our results suggest that the sialic acid binding activity facilitates the infection by TGEV under unfavorable environmental conditions. The dependence on the sialic acid binding activity for an efficient infection differs in the analyzed TGEV strains.</p

Spleen tyrosine kinase mediates innate and adaptive immune crosstalk in SARS-CoV-2 mRNA vaccination

Durable cell-mediated immune responses require efficient innate immune signaling and the release of pro-inflammatory cytokines. How precisely mRNA vaccines trigger innate immune cells for shaping antigen specific adaptive immunity remains unknown. Here, we show that SARS-CoV-2 mRNA vaccination primes human monocyte-derived macrophages for activation of the NLRP3 inflammasome. Spike protein exposed macrophages undergo NLRP3-driven pyroptotic cell death and subsequently secrete mature interleukin-1β. These effects depend on activation of spleen tyrosine kinase (SYK) coupled to C-type lectin receptors. Using autologous cocultures, we show that SYK and NLRP3 orchestrate macrophage-driven activation of effector memory T cells. Furthermore, vaccination-induced macrophage priming can be enhanced with repetitive antigen exposure providing a rationale for prime-boost concepts to augment innate immune signaling in SARS-CoV-2 vaccination. Collectively, these findings identify SYK as a regulatory node capable of differentiating between primed and unprimed macrophages, which modulate spike protein-specific T cell responses

ARSB IS REQUIRED FOR BONE REMODELING

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Antiskyrmions and their electrical footprint in crystalline mesoscale structures of Mn1.4PtSn

Skyrmionic materials hold the potential for future information technologies, such as racetrack memories. Key to that advancement are systems that exhibit high tunability and scalability, with stored information being easy to read and write by means of all-electrical techniques. Topological magnetic excitations such as skyrmions and antiskyrmions, give rise to a characteristic topological Hall effect. However, the electrical detection of antiskyrmions, in both thin films and bulk samples has been challenging to date. Here, we apply magneto-optical microscopy combined with electrical transport to explore the antiskyrmion phase as it emerges in crystalline mesoscale structures of the Heusler magnet Mn1.4PtSn. We reveal the Hall signature of antiskyrmions in line with our theoretical model, comprising anomalous and topological components. We examine its dependence on the vertical device thickness, field orientation, and temperature. Our atomistic simulations and experimental anisotropy studies demonstrate the link between antiskyrmions and a complex magnetism that consists of competing ferromagnetic, antiferromagnetic, and chiral exchange interactions, not captured by micromagnetic simulations

A Novel DBL-Domain of the P. falciparum 332 Molecule Possibly Involved in Erythrocyte Adhesion

Plasmodium falciparum malaria is brought about by the asexual stages of the parasite residing in human red blood cells (RBC). Contact between the erythrocyte surface and the merozoite is the first step for successful invasion and proliferation of the parasite. A number of different pathways utilised by the parasite to adhere and invade the host RBC have been characterized, but the complete biology of this process remains elusive. We here report the identification of an open reading frame (ORF) representing a hitherto unknown second exon of the Pf332 gene that encodes a cysteine-rich polypeptide with a high degree of similarity to the Duffy-binding-like (DBL) domain of the erythrocyte-binding-ligand (EBL) family. The sequence of this DBL-domain is conserved and expressed in all parasite clones/strains investigated. In addition, the expression level of Pf332 correlates with proliferation efficiency of the parasites in vitro. Antibodies raised against the DBL-domain are able to reduce the invasion efficiency of different parasite clones/strains. Analysis of the DBL-domain revealed its ability to bind to uninfected human RBC, and moreover demonstrated association with the iRBC surface. Thus, Pf332 is a molecule with a potential role to support merozoite invasion. Due to the high level of conservation in sequence, the novel DBL-domain of Pf332 is of possible importance for development of novel anti-malaria drugs and vaccines

Depletion of WFS1 compromises mitochondrial function in hiPSC-derived neuronal models of Wolfram syndrome

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