The mitochondrial protein MAVS (also known as IPS-1, VISA, and CARDIF) interacts with RIG-I-like receptors (RLRs) to induce type I interferon (IFN-I). NLRX1 is a mitochondrial nucleotide-binding, ...leucine-rich repeats (NLR)-containing protein that attenuates MAVS-RLR signaling. Using Nlrx1−/− cells, we confirmed that NLRX1 attenuated IFN-I production, but additionally promoted autophagy during viral infection. This dual function of NLRX1 paralleled the previously described functions of the autophagy-related proteins Atg5-Atg12, but NLRX1 did not associate with Atg5-Atg12. High-throughput quantitative mass spectrometry and endogenous protein-protein interaction revealed an NLRX1-interacting partner, mitochondrial Tu translation elongation factor (TUFM). TUFM interacted with Atg5-Atg12 and Atg16L1 and has similar functions as NLRX1 by inhibiting RLR-induced IFN-I but promoting autophagy. In the absence of NLRX1, increased IFN-I and decreased autophagy provide an advantage for host defense against vesicular stomatitis virus. This study establishes a link between an NLR protein and the viral-induced autophagic machinery via an intermediary partner, TUFM.
► NLRX1 suppresses VSV-mediated type 1 IFN production but enhances autophagy ► TUFM works with NLRX1 to inhibit RLR-induced type 1 IFN signaling ► TUFM enhances autophagy and interacts with Atg5-Atg12 and Atg16L1 ► NLRX1-mediated autophagy and IFN-I inhibition enhance VSV replication
Spermatogenesis requires intricate interactions between the germline and somatic cells. Within a given cross section of a seminiferous tubule, multiple germ and somatic cell types co-occur. This ...cellular heterogeneity has made it difficult to profile distinct cell types at different stages of development. To address this challenge, we collected single-cell RNA sequencing data from ∼35,000 cells from the adult mouse testis and identified all known germ and somatic cells, as well as two unexpected somatic cell types. Our analysis revealed a continuous developmental trajectory of germ cells from spermatogonia to spermatids and identified candidate transcriptional regulators at several transition points during differentiation. Focused analyses delineated four subtypes of spermatogonia and nine subtypes of Sertoli cells; the latter linked to histologically defined developmental stages over the seminiferous epithelial cycle. Overall, this high-resolution cellular atlas represents a community resource and foundation of knowledge to study germ cell development and in vivo gametogenesis.
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•Analysis of ∼35,000 cells identifies known and unexpected mouse testicular cell types•Germ cell development includes discrete states followed by a continuous trajectory•Differential gene expression identifies new regulators of spermatogenesis•Four spermatogonia subtypes and nine Sertoli cell subtypes map to known stages
Male germ cell development requires intricate interplay between mitotic, meiotic, and differentiating germ cells and supporting somatic cells. Green et al. apply single-cell RNA sequencing to generate a mouse spermatogenesis resource to untangle cellular heterogeneity within the adult testis, identify regulators of germ cell differentiation, and discover somatic cell types.
The body is exposed to foreign pathogens every day, but remarkably, most pathogens are effectively cleared by the innate immune system without the need to invoke the adaptive immune response. Key ...cellular components of the innate immune system include macrophages and neutrophils and the recruitment and function of these cells are tightly regulated by chemokines and cytokines in the tissue space. Innate immune responses are also known to regulate development of adaptive immune responses often via the secretion of various cytokines. In addition to these protein regulators, numerous lipid mediators can also influence innate and adaptive immune functions. In this review, we cover one particular lipid regulator, prostaglandin E
(PGE
) and describe its synthesis and signaling and what is known about the ability of this lipid to regulate immunity and host defense against viral, fungal and bacterial pathogens.
Organotypic slice cultures of brain or spinal cord have been a longstanding tool in neuroscience research but their utility for understanding Alzheimer's disease (AD) and other neurodegenerative ...proteinopathies has only recently begun to be evaluated. Organotypic brain slice cultures (BSCs) represent a physiologically relevant three-dimensional model of the brain. BSCs support all the central nervous system (CNS) cell types and can be produced from brain areas involved in neurodegenerative disease. BSCs can be used to better understand the induction and significance of proteinopathies underlying the development and progression of AD and other neurodegenerative disorders, and in the future may serve as bridging technologies between cell culture and in vivo experiments for the development and evaluation of novel therapeutic targets and strategies. We review the initial development and general use of BSCs in neuroscience research and highlight the advantages of these cultures as an ex vivo model. Subsequently we focus on i) BSC-based modeling of AD and other neurodegenerative proteinopathies ii) use of BSCs to understand mechanisms underlying these diseases and iii) how BSCs can serve as tools to screen for suitable therapeutics prior to in vivo investigations. Finally, we will examine i) open questions regarding the use of such cultures and ii) how emerging technologies such as recombinant adeno-associated viruses (rAAV) may be combined with these models to advance translational research relevant to neurodegenerative disorders.
This introduction to the special issue considers past and current research on “Social Capital and Entrepreneurship” to develop a schema and an associated research agenda. With the general goal of ...establishing social capital as a foundational theory of entrepreneurship, we discuss how future research can utilize social capital perspectives across levels of analysis and contexts to explain a wide variety of entrepreneurship phenomena.
Fibrinolysis was initially defined using rapid thrombelastography (rTEG). The cutoffs for the pathologic extremes of the fibrinolytic system, hyperfibrinolysis and shutdown, were both defined based ...on association with mortality. We propose to redefine these phenotypes for both TEG and for rotational thrombelastometry, the other commonly used viscoelastic assay.
Rotational thrombelastometry, rTEG, and clinical data were prospectively collected on trauma patients admitted to an urban Level I trauma center from 2010 to 2016. Hyperfibrinolysis was defined as the Youden index from EXTEM-clot lysis index 60 minutes after clotting time (CLI60) and rTEG-fibrinolysis 30 minutes after achieving MA (LY30) for predicting massive transfusion (>10 red blood cell units, or death per 6 hours after injury) as a surrogate for severe bleeding. Patients identified as having hyperfibrinolysis were then removed from the data set, and the cutoff for fibrinolysis shutdown was derived as the optimal cutoff for predicting mortality in the remaining patients.
Overall, 216 patients (median age, 36 years (interquartile range, 27-49 years), 82% men, 58% blunt injury) were included. Of these, 16% required massive transfusion, and 12.5% died. Rapid thrombelastography phenotypes were redefined as hyperfibrinolysis: rTEG-LY30 greater than7.7%, physiologic rTEG-LY30 0.6% to7.6%, and shutdown rTEG-LY30 less than 0.6%. EXTEM-CLI60 fibrinolysis phenotypes were hyperfibrinolysis CLI60 less than 82%, physiologic (CLI60, 82-97.9%), and shutdown (CLI60 > 98%). Weighted kappa statistics revealed moderate agreement between rotational thrombelastometry- and rTEG-defined fibrinolysis (k = 0.51; 95% confidence interval, 0.39-0.63), with disagreement mostly in the shutdown and physiologic categories.
We confirmed the U-shaped distribution of death related to fibrinolysis system abnormalities. Both rTEG LY30 and EXTEM CLI60 can identify the spectrum of fibrinolytic phenotypes, have moderate agreement, and can be used to guide hemostatic resuscitation.
Diagnostic study, level III.
Using the atomic structures of the large ribosomal subunit from Haloarcula marismortui and its complexes with two substrate analogs, we establish that the ribosome is a ribozyme and address the ...catalytic properties of its all-RNA active site. Both substrate analogs are contacted exclusively by conserved ribosomal RNA (rRNA) residues from domain V of 23S rRNA; there are no protein side-chain atoms closer than about 18 angstroms to the peptide bond being synthesized. The mechanism of peptide bond synthesis appears to resemble the reverse of the acylation step in serine proteases, with the base of A2486 (A2451 in Escherichia coli) playing the same general base role as histidine-57 in chymotrypsin. The unusual pKa(where Kais the acid dissociation constant) required for A2486 to perform this function may derive in part from its hydrogen bonding to G2482 (G2447 in E. coli), which also interacts with a buried phosphate that could stabilize unusual tautomers of these two bases. The polypeptide exit tunnel is largely formed by RNA but has significant contributions from proteins L4, L22, and L39e, and its exit is encircled by proteins L19, L22, L23, L24, L29, and L31e.
Stabilized Criegee intermediates (SCI) are formed during the ozonolysis of unsaturated hydrocarbons and have been implicated in the formation of hydroxyl radicals and aerosols. Previous theoretical ...research S. Jørgenson and A. Gross, J. Phys. Chem. A, 2009, 113, 10284-10290 computed the rate constants for addition of ammonia to simple SCIs, but reported a wide distribution of quantum chemical energies, depending on the basis set used. We report optimized geometries for these reactions at the CCSD(T)/ANO2 and CCSD(T)/ANO1 levels, and CCSD(T)/CBS energies with perturbative quadruples corrections. We find the inclusion of perturbative quadruples corrections elevates the energy of the transition state by 0.76-0.88 kcal mol
relative to the reactants, which qualitatively changes the reaction surface. We calculate rate constants and find that Jørgenson and Gross previously overestimated the rate constants for ammonia addition to SCIs, but were within an order of magnitude. This supports the previous conclusion of Vereecken et al. L. Vereecken, H. Harder and A. Novelli, Phys. Chem. Chem. Phys., 2012, 14, 14682-14695 that ammonia addition to SCIs is a negligible sink of Criegee intermediates.
Viscoelastic assays, such as thrombelastography (TEG) and rotational thrombelastometry (ROTEM), have emerged as point-of-care tools that can guide the hemostatic resuscitation of bleeding injured ...patients. This article describes the role of TEG in contemporary trauma care by explaining this assay's methodology, clinical applications, and result interpretation through description of supporting studies to provide the reader with an evidence-based user's guide. Although TEG and ROTEM are assays based on the same viscoelastic principle, this article is focused on data supporting the use of TEG in trauma, because it is available in trauma centers in North America; ROTEM is mostly available in Europe.
In the wake of the COVID-19 pandemic, social restrictions to contain the spread of the virus have disrupted behaviors across the 24-h day including physical activity, sedentary behavior, and sleep ...among children (5–12 years old) and adolescents (13–17 years old). Preliminary evidence reports significant decreases in physical activity, increases in sedentary behavior, and disrupted sleep schedules/sleep quality in children and adolescents. This commentary discusses the impact of COVID-19-related restrictions on behaviors across the 24-h day in children and adolescents. Furthermore, we suggest recommendations through the lens of a socio-ecological model to provide strategies for lasting behavior change to insure the health and well-being of children and adolescents during the COVID-19 pandemic.
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