bluuebunny/crossref_metadata_2025_split huggingface dataset:
The SARS-CoV-2 Spike mutation D614G increases entry fitness across a range of ACE2 levels, directly outcompetes the wild type, and is preferentially incorporated into trimers
Authors: Michaud William A. , Boland Genevieve M. , Rabi S. Alireza 
Published: August 2020

AbstractEarly in the current pandemic, the D614G mutation arose in the Spike protein of SARS-CoV-2 and quickly became the dominant variant globally. Mounting evidence suggests D614G enhances viral entry. Here we use a direct competition assay with single-cycle viruses to show that D614G outcompetes the wildtype. We developed a cell line with inducible ACE2 expression to confirm that D614G more efficiently enters cells with ACE2 levels spanning the different primary cells targeted by SARS-CoV-2. Using a new assay for crosslinking and directly extracting Spike trimers from the pseudovirus surface, we found an increase in trimerization efficiency and viral incorporation of D614G protomers. Our findings suggest that D614G increases infection of cells expressing a wide range of ACE2, and informs the mechanism underlying enhanced entry. The tools developed here can be broadly applied to study other Spike variants and SARS-CoV-2 entry, to inform functional studies of viral evolution and vaccine development.
Dual RNAseq highlights the kinetics of skin microbiome and fish host responsiveness to bacterial infection
Authors: Le Luyer J. , Schull Q. , Auffret P. , lopez P. , Crusot M. , Belliard C. , Basset C. , Carradec Q. , Poulain J. , Planes S. , Saulnier D. 
Published: August 2020

Abstracta)BackgroundTenacibaculum maritimumis a worldwide-distributed fish pathogen known for causing dramatic damages on a broad range of wild and farmed marine fish populations. Recently sequenced genome ofT. maritimumstrain NCIMB 2154Tprovided unprecedented information on the possible molecular mechanisms involved in virulence for this species. However, little is known on the dynamic on the infectionin vivo, and information are lacking on both the intrinsic host response (gene expression) and its associated microbiome community. Here, we applied complementary omic approaches, including dual RNAseq and 16S rRNA gene metabarcoding sequencing using Nanopore and short-reads Illumina technologies to unravel the host-pathogens interplay in experimental infection system using the tropical fishPlatax orbicularisas model.b)ResultsWe show thatT. maritimumtranscriptomic landscape during infection is characterized by an enhancement of antibiotic catalytic and glucan catalytic functions while decreasing specific sulphate assimilation process, compared toin vitrocultures. Simultaneously, fish host display a large palette of immune effectors, notably involving innate response and triggering acute inflammatory response. In addition, results suggest that fish activate adaptive immune response visible through stimulation of T-helper cells, Th17, with congruent reduction of Th2 and T-regulatory cells. Fish were however largely sensitive to infection, and less than 25% of them survived after 96hpi. These surviving fish showed no evidence of stress (cortisol levels) as well as no significant difference in microbiome diversity compared to control at the same sampling time. The presence ofTenacibaculumin resistant fish skin and the total absence of any skin lesion suggest that these fish did not escape contact with the pathogen but rather prevent the pathogen entry. In these individuals we detected the up-regulation of specific immune-related genes differentiating resistant from control at 96hpi, which suggests a possible genomic basis of resistance while no genetic variations in coding regions was reported.c)ConclusionHere we refine the interplay between common fish pathogens and host immune response during experimental infection. We further highlight key actors of defense response, pathogenicity and possible genomic bases of resistance toT. maritimum.
Natural resistance to worms exacerbates bovine tuberculosis severity independently of worm coinfection
Authors: Ezenwa Vanessa O. , Budischak Sarah A. , Buss Peter , Seguel Mauricio , Luikart Gordon , Jolles Anna E. , Sakamoto Kaori 
Published: August 2020

AbstractPathogen interactions arising during coinfection can exacerbate disease severity, for example, when the immune response mounted against one pathogen negatively affects defense of another. It is also possible that host immune responses to a pathogen shaped by historical evolutionary interactions between host and pathogen, may modify host immune defenses in ways that have repercussions for other pathogens. In this case, negative interactions between two pathogens could emerge even in the absence of concurrent infection. Parasitic worms and tuberculosis (TB) are involved in one of the most geographically extensive of pathogen interactions, and during coinfection, worms can exacerbate TB disease outcomes. Here, we show that in a wild mammal, natural resistance to worms affects bovine tuberculosis (BTB) severity independently of active worm infection. We found that worm-resistant individuals were more likely to die of BTB than were non-resistant individuals, and their disease progressed more quickly. Anthelmintic treatment moderated, but did not eliminate, the resistance effect, and the effects of resistance and treatment were additive with untreated, resistant individuals experiencing the highest mortality. Interestingly, resistance and anthelmintic treatment had non-overlapping effects on BTB pathology. The effects of resistance manifested in the lungs (the primary site of BTB infection), while the effects of treatment manifested almost entirely in the lymph nodes (the site of disseminated disease), suggesting that resistance and active worm infection affect BTB progression via distinct mechanisms. Our findings reveal that interactions between pathogens can occur as a consequence of processes arising on very different timescales.
Heterologous expression of cryptomaldamide in a cyanobacterial host
Authors: Taton Arnaud , Ecker Andrew , Diaz Brienna , Moss Nathan A. , Anderson Brooke , Reher Raphael , Leão Tiago F. , Simkovsky Ryan , Dorrestein Pieter C. , Gerwick Lena , Gerwick William H. , Golden James W. 
Published: August 2020

ABSTRACTFilamentous marine cyanobacteria make a variety of bioactive molecules that are produced by polyketide synthases, non-ribosomal peptide synthetases, and hybrid pathways that are encoded by large biosynthetic gene clusters. These cyanobacterial natural products represent potential drugs leads; however, thorough pharmacological investigations have been impeded by the limited quantity of compound that is typically available from the native organisms. Additionally, investigations of the biosynthetic gene clusters and enzymatic pathways have been difficult due to the inability to conduct genetic manipulations in the native producers. Here we report a set of genetic tools for the heterologous expression of biosynthetic gene clusters in the cyanobacteriaSynechococcus elongatusPCC 7942 andAnabaena(Nostoc) PCC 7120. To facilitate the transfer of gene clusters in both strains, we engineered a strain ofAnabaenathat containsS. elongatushomologous sequences for chromosomal recombination at a neutral site and devised a CRISPR-based strategy to efficiently obtain segregated double recombinant clones ofAnabaena. These genetic tools were used to express the large 28.7 kb cryptomaldamide biosynthetic gene cluster from the marine cyanobacteriumMoorena(Moorea)producensJHB in both model strains.S. elongatusdid not produce cryptomaldamide, however high-titer production of cryptomaldamide was obtained inAnabaena. The methods developed in this study will facilitate the heterologous expression of biosynthetic gene clusters isolated from marine cyanobacteria and complex metagenomic samples.Abstract Figure
Aberrant DJ-1 expression underlies L-type calcium channel hypoactivity in tuberous sclerosis complex and Alzheimer’s disease
Authors: Niere Farr , Uneri Ayse , McArdle Colin J. , Cacheaux Luisa P. , Egido-Betancourt Hailey X. , Namjoshi Sanjeev , Reynoldson Cameron , Penaranda Juan , Wang Xin , Taylor William C. , Craft Suzanne , Keene Christopher Dirk , Ma Tao , Raab-Graham Kimberly F. 
Published: August 2020

AbstractL-type voltage-dependent Ca2+channels (L-VDCC) integrate synaptic signals to facilitate a plethora of cellular mechanisms. L-VDCC dysfunction is implicated in several neurological and psychiatric diseases. Despite their importance, signals upstream of L-VDCC activity that regulate their channel density, however, are poorly defined. In disease models with overactive mammalian target of rapamycin complex 1 (mTORC1) signaling (or mTORopathies), including tuberous sclerosis (TS) and Alzheimer’s disease (AD), we report a novel mechanism downstream of mTORC1 signaling that results in a deficit in dendritic L-VDCC activity. Deficits in L-VDCC activity are associated with increased expression of the mTORC1-regulated RNA-binding protein DJ-1. DJ-1 binds the mRNA coding the auxiliary Ca2+channel subunit α2δ2 responsible for shuttling L-VDCC to the membrane and represses its expression. Moreover, this novel DJ-1/α2δ2/L-VDCC pathway is disrupted in human AD and preclinical models of AD and TS. Our discovery that DJ-1 directs L-VDCC activity and L-VDCC-associated protein α2δ2 at the synapse suggests that DJ-1/α2δ2/L-VDCC is a common, fundamental pathway disrupted in TS and AD that can be targeted in clinical mTORopathies.Significance StatementMany neurological disorders share symptoms, despite disparity among diseases. Treatments are prescribed based on diagnosis rather than individual symptoms. While only treating symptoms may obscure the disease, mechanism-based drug development allows the two approaches to converge. Hub proteins, those that coordinate the expression of proteins that mediate specific cellular functions, may be dysregulated across a broad range of disorders. Herein, we show that the RNA-binding protein DJ-1 controls the activity of L-type voltage-dependent calcium channels (L-VDCC), via the expression of its auxiliary subunit alpha2delta2 (α2δ2). Importantly, we demonstrate that this novel DJ-1/α2δ2/L-VDCC pathway is commonly disrupted among neurological disorders, namely Alzheimer’s disease (AD) and Tuberous Sclerosis (TS). Collectively, these data rationalize mechanism-based drug therapy to treat disease.
Specific decorations of 17-hydroxygeranyllinalool diterpene glycosides solve the autotoxicity problem of chemical defense inNicotiana attenuata
Authors: Heiling Sven , Llorca Lucas Cortes , Li Jiancai , Gase Klaus , Schmidt Axel , Schäfer Martin , Schneider Bernd , Halitschke Rayko , Gaquerel Emmanuel , Baldwin Ian Thomas 
Published: August 2020

Abstract17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) are abundant and potent anti-herbivore defense metabolites inNicotiana attenuatawhose glycosylation and malonylation biosynthetic steps are regulated by jasmonate signaling. To characterize the biosynthetic pathway of HGL-DTGs, we conducted a genome-wide analysis of uridine diphosphate glycosyltransferases (UGTs) and identified 107 members of family-1 UGTs. Tissue-specific time-course transcriptional profiling revealed that the transcripts of three UGTs were highly correlated with two HGL-DTG key biosynthetic genes: geranylgeranyl diphosphate synthase (NaGGPPS)and geranyllinalool synthase (NaGLS). NaGLS’s role in HGL-DTG biosynthesis was confirmed by virus-induced gene-silencing. Silencing the UDP-rhamnosyltransferase,UGT91T1,indicated its role in the rhamnosylation of HGL-DTGs.In vitroenzyme assays revealed that UGT74P3 and UGT74P4 use UDP-glucose for the glucosylation of 17-hydroxygeranyllinalool (17-HGL) to lyciumoside I.UGT74P3andUGT74P5 stablysilenced plants were severely developmentally deformed, suggesting a phytotoxic effect of 17-HGL. Applications of synthetic 17-HGL and silencing of these UGTs in HGL-DTG-free plants confirmed the phytotoxic effect of 17-HGL. Feeding assays withManduca sextalarvae revealed the defensive functions of the glucosylation and rhamnosylation steps in HGL-DTG biosynthesis. Glucosylation is a critical step that contributes to the metabolites’ defensive function and solves the autotoxicity problem of this potent chemical defense.
Low sperm to egg ratio required for successful in vitro fertilisation in a pair-spawning teleost, Senegalese sole (Solea senegalensis)
Authors: Ramos-Júdez Sandra , González-López Wendy Ángela , Ostos Jhons Huayanay , Mamani Noemí Cota , Alemán Carlos Marrero , Beirão José , Duncan Neil 
Published: August 2020

AbstractCultured Senegalese sole (Solea senegalensis) breeders fail to spawn fertilised eggs and this bottleneck could be solved with the implementation of large-scalein vitrofertilisation protocols. However, low production of poor-quality sperm has frustrated the development ofin vitrofertilisation protocols. Cultured females were induced to ovulate with a 5 µg kg-1single injection of gonadotropin releasing hormone agonist (GnRHa) and good quality eggs (82.6 ± 9.2% fertilisation) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilise the eggs. A non-linear regression, an exponential rise to a maximum (R = 0.93, P < 0.0001) described the number of motile spermatozoa required to fertilise a viable egg and 1617 motile spermatozoa were sufficient to fertilise 99 ± 12% (± 95% CI) of viable eggs. Similar, spermatozoa egg-1ratios of 592 ± 611 motile spermatozoa egg-1were used in large-scalein vitrofertilisations with 190,512 ± 38,471 eggs. The sperm from a single male (145 ± 50 µL or 8.0 ± 6.8 × 108spermatozoa) was used to fertilise the eggs. The mean hatching rate of the large-scalein vitrofertilisations was 70 ± 14 % to provide 131,540 ± 34,448 larvae per fertilisation. When unfertilised eggs were stored at room temperature the percentage of viable eggs decreased gradually and indicated the sooner eggs were fertilised after stripping the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2 %) compared to dilution in modified Leibovitz immediately after collection (6.6 ± 4.9 %). Senegalese sole have a pair-spawning reproductive behaviour characterised by external gamete fertilisation in close proximity with no sperm competition. The low spermatozoa egg-1ratio required for maximum fertilisation was consistent with this reproductive behaviour and strategy. The provision of a large-scalein vitrofertilisation protocol (200 µL of sperm per 100 mL of eggs) will enable the industry to operate sustainably and implement breeding programs to improve production.
Tetanus neurotoxin sensitive SNARE-mediated glial signaling limits motoneuronal excitability
Authors: Böhme Mathias A. , McCarthy Anthony W. , Berezeckaja Monika , Ponimaskin Kristina , Walter Alexander M. 
Published: August 2020

AbstractPeripheral nerves contain motoneuron axons coated by glial cells, which essentially contribute to function but cellular reactions remain poorly understood. We here identify non-neuronal Synaptobrevin (Syb) as the essential vesicular SNARE in glia to insulate and metabolically supplyDrosophilamotoneurons. Interfering with Syb-functionality by glial knockdown, or glial expression of tetanus neurotoxin light chain (TeNT-LC) caused motonerve disintegration, blocked axonal transport, induced tetanic muscle hyperactivity and caused lethal paralysis. Surprisingly, not the established TeNT-LC-target, neuronal Synaptobrevin (nSyb), is the relevant SNARE, but non-neuronal Synaptobrevin (Syb): Knockdown of Syb- (but not nSyb-) phenocopied glial TeNT-LC expression whose effects were reverted by a TeNT-LC-insensitive Syb mutant. We link Syb-necessity to two distinct glia: to establish nerve insulating septate junctions in subperineurial glia and to integrate monocarboxylate transporters along the nerve in wrapping glia for motoneuronal metabolic supply. Our study identifies crucial roles of Syb in glial subtypes for nerve function and pathology, animal motility and survival.
Antibody-based delivery of Interleukin-9 to neovascular structures: therapeutic evaluation in cancer and arthritis
Authors: Gouyou Baptiste , Ongaro Tiziano , Cazzamalli Samuele , Luca Roberto De , Kerschenmeyer Anne , Valet Philippe , Villa Alessandra , Neri Dario , Matasci Mattia 
Published: August 2020

ABSTRACTInterleukin-9 (IL9) is a cytokine with multiple functions, including the ability to activate group 2 innate lymphoid cells (ILC2s), which has been postulated to be therapeutically active in mouse models of arthritis. Similarly, IL9 has been suggested to play an important role in tumor immunity. Here, we describe the cloning, expression and characterization of three fusion proteins based on murine IL9 and the F8 antibody, specific to the alternatively-spliced EDA domain of fibronectin. EDA is strongly expressed in cancer and in various arthritic conditions, while being undetectable in the majority of healthy organs. IL9-based fusion proteins with an irrelevant antibody specific to hen egg lysozyme served as negative control in our study. The fusion proteins were characterized by quantitative biodistribution analysis in tumor-bearing mice using radioiodinated protein preparations. The highest tumor uptake and best tumor:organ ratios were observed for a format, in which the IL9 moiety was flanked by two units of the F8 antibody in single-chain Fv format. Biological activity of IL9 was retained when the payload was fused to antibodies. However, the targeted delivery of IL9 to the disease site resulted in a modest anti-tumor activity in three different murine models of cancer (K1735M2, CT26 and F9), while no therapeutic benefit was observed in a collagen induced model of arthritis. Collectively, these results confirm the possibility to deliver IL9 to the site of disease but cast doubts about the alleged therapeutic activity of this cytokine in cancer and arthritis, which has been postulated in previous publications.
Prenatal Stress Dysregulates Resting-State Functional Connectivity and Sensory Motifs
Authors: Rezaei Zahra , Jafari Zahra , Afrashteh Navvab , Torabi Reza , Singh Surjeet , Kolb Bryan E. , Davidsen Jörn , Mohajerani Majid H. 
Published: August 2020

AbstractPrenatal stress (PS) can impact fetal brain structure and function and contribute to higher vulnerability to neurodevelopmental and neuropsychiatric disorders. To understand how PS alters evoked and spontaneous neocortical activity and intrinsic brain functional connectivity, mesoscale voltage imaging was performed in adult C57BL/6NJ mice that had been exposed to auditory stress on gestational days 12-16, the age at which neocortex is developing. PS mice had a four-fold higher basal corticosterone level and reduced amplitude of cortical sensory-evoked responses to visual, auditory, whisker, forelimb, and hindlimb stimuli. Relative to control animals, PS also led to a general reduction of resting-state functional connectivity, as well as reduced inter-modular connectivity, enhanced intra-modular connectivity, and altered frequency of auditory and forelimb spontaneous sensory motifs. These resting-state changes resulted in a cortical connectivity pattern featuring disjoint but tight modules and a decline in network efficiency. The findings demonstrate that cortical connectivity is sensitive to PS and exposed offspring may be at risk for adult stress-related neuropsychiatric disorders.
Engineering murine GITRL for antibody-mediated delivery to tumor-associated blood vessels
Authors: Mock Jacqueline , Rascon Itzel Astiazaran , Stringhini Marco , Catalano Marco , Neri Dario 
Published: August 2020

ABSTRACTPreclinical evidence has suggested that the Glucocorticoid-Induced TNFR-related protein (GITR) may be valuable a target for the development of anticancer therapeutics, but clinical studies with GITR ligand (GITRL) have been disappointing. Here, we report the development of a fusion protein featuring GITR ligand (GITRL) fused to the F8 antibody which targets the alternatively-spliced EDA domain of fibronectin, a tumor-associated antigen often found around the tumor neovasculature. Five different formats for F8-GITRL fusion proteins were cloned and characterized, but quantitative biodistribution studies failed to evidence a preferential accumulation at the tumor site. Thein vivotumor targeting properties of F8-GITRL could be substantially improved by enzymatic deglycosylation or site-directed mutagenesis of theN-glycosylation consensus sequence. However, therapy studies in a murine model of cancer with the glycoengineered F8-GITRL N74S and N157T variant failed to elicit a durable anti-tumor response, both in monotherapy and in combination with PD-1 blockade.HIGHLIGHTSDifferent formats of fusion proteins featuring Glucocorticoid-induced TNFR-related protein ligand (GITRL) fused to a tumor-targeting antibody were produced.The tumor uptake of the fusion proteins could be increased by enzymatic deglycosylation of the fusion protein or by site-directed mutagenesis of theN-glycosylation consensus sequences.The fusion protein developed in this study failed to show any anti-tumor activity either alone or in combination with PD-1 inhibition.
Large-Scale Transcriptional Profiling of Molecular Perturbations Reveals Cell Type Specific Responses and Implications for Environmental Screening
Authors: Zhang Kun , Zhao Yanbin 
Published: August 2020

AbstractCell-based assays represent nearly half of all high-throughput screens currently conducted for risk assessment of environmental chemicals. However, the sensitivity and heterogeneity among cell lines has long been concerned but explored only in a limited manner. Here, we address this question by conducting a large scale transcriptomic analysis of the responses of discrete cell lines to specific small molecules. Our results illustrate heterogeneity of the extent and timing of responses among cell lines. Interestingly, high sensitivity and/or heterogeneity was found to be cell type-specific or universal depending on the different mechanism of actions of the compounds. Our data provide a novel insight into the understanding of cell-small molecule interactions and have substantial implications for the design, execution and interpretation of high-throughput screening assays.
Corticosteroids and cellulose purification improve respectively the in vivo translation and vaccination efficacy of self-amplifying mRNAs
Authors: Zhong Zifu , Cafferty Séan Mc , Opsomer Lisa , Wang Haixiu , Huysmans Hanne , De Temmerman Joyca , Lienenklaus Stefan , Portela Catani João Paulo , Combes Francis , Sanders Niek N. 
Published: August 2020

AbstractSynthetic mRNAs are an appealing therapeutic platform with multiple biomedical applications ranging from protein replacement therapy to vaccination. In comparison to conventional mRNA, synthetic self-amplifying mRNAs (sa-mRNAs) are gaining increased interest due to their higher and longer-lasting expression. However, sa-mRNAs also elicit an innate immune response, which may complicate the clinical translation of this platform. Approaches to reduce the innate immunity of sa-mRNAs have not been studied in detail. In this work we investigated the effect of several innate immune inhibitors and a novel cellulose-based mRNA purification approach on the type I interferon (IFN) response, translation and vaccination efficacy of our formerly developed sa-mRNA vaccine against Zika virus. Among the investigated inhibitors, we found that topical application of clobetasol at the sa-mRNA injection site was the most efficient in suppressing the type I IFN response and increasing the translation of sa-mRNA. However, clobetasol prevented the formation of antibodies against sa-mRNA encoded antigens and should therefore be avoided in a vaccination context. Residual dsRNA by-products of the in vitro transcription reaction are known inducers of immediate type I IFN responses. We additionally demonstrate drastic reduction of these dsRNA by-products upon cellulose-based purification, consequently reducing the innate immune response and improving sa-mRNA vaccination efficacy.