bluuebunny/crossref_metadata_2025_split huggingface dataset:
High-throughput phenotypic screen identifies a new family of potent anti-amoebic compounds
Authors: Sauvey Conall , Ehrenkaufer Gretchen , Blevitt Jonathan , Jackson Paul , Abagyan Ruben 
Published: October 2021

AbstractEntamoeba histolytica is a disease-causing parasitic amoeba which affects an estimated 50 million people worldwide, particularly in socioeconomically vulnerable populations experiencing water sanitation issues. Infection with E. histolytica is referred to as amoebiasis, and can cause symptoms such as colitis, dysentery, and even death in extreme cases. Drugs exist that are capable of killing this parasite, but they are hampered by downsides such as significant adverse effects at therapeutic concentrations, issues with patient compliance, the need for additional drugs to kill the transmissible cyst stage, and potential development of resistance. Past screens of small and medium sized chemical libraries have yielded anti-amoebic candidates, thus rendering high-throughput screening a promising direction for new drug discovery in this area. In this study, we screened a curated 80,000-compound library from Janssen pharmaceuticals against E. histolytica trophozoites in vitro, and from it identified a highly potent new inhibitor compound. Further experimentation confirmed the activity of this compound, as well as that of several structurally related compounds, originating from both the Janssen Jump-stARter library, and from chemical vendors, thus highlighting a new structure-activity relationship (SAR). In addition, we confirmed that the compound inhibited E. histolytica survival as rapidly as the current standard of care and inhibited transmissible cysts of the related model organism Entamoeba invadens. Together these results constitute the discovery of a novel class of chemicals with favorable in vitro pharmacological properties which may lead to an improved therapy against this parasite and in all of its life stages.Author summaryThe parasite Entamoeba histolytica represents a significant challenge in the field of global health. It currently infects and causes disease among millions of people worldwide, particularly those lacking access to clean water. Drugs exist to treat this disease, but nevertheless it persists as a problem, likely at least partly due to problems and downsides inherent to these drugs. Hence the search for new and better ones is needed. We report here our contribution to this search, consisting of testing a large, carefully-curated collection of tens of thousands of chemicals for their ability to kill E. histolytica. This large-scale test resulted in the identification of one of the compounds as potently anti-amoebic, capable of killing the parasite cells at extremely low concentrations. Further experimentation found several chemically-related compounds to also possess this property, and additionally found the first compound capable of killing the infective life stage of another Entamoeba parasite. These results have revealed an entire new family of chemicals with good potential for development as better drugs against this disease.
Integrative analysis identified common and unique molecular signatures in hepatobiliary cancers
Authors: Roy Nabanita , Lodh Ria , Sarma Anupam , Bhattacharyya Dhruba Kumar , Barah Pankaj 
Published: October 2021

AbstractHepatobiliary cancers (HBCs) are the most aggressive and sixth most diagnosed cancers globally. Biomarkers for timely diagnosis and targeted therapy in HBCs are still limited. Considering the gap, our objective is to identify unique and overlapping molecular signatures associated with HBCs. We analyzed publicly available transcriptomic datasets on Gallbladder cancer (GBC), Hepatocellular carcinoma (HCC), and Intrahepatic cholangiocarcinoma (ICC) to identify potential biomarkers using integrative systems approaches. An effective Common and Unique Molecular Signature Identification (CUMSI) approach has been developed, which contains analysis of differential gene expression (DEG), gene co-expression networks (GCN), and protein-protein interactions (PPIs) networks. Functional analysis of the DEGs unique for GBC, HCC, and ICC indicated that GBC is associated with cellular processes, HCC is associated with immune signaling pathways, and ICC is associated with lipid metabolic pathways. Our findings shows that the hub genes and pathways identified for each individual cancer type of the HBS are related with the primary function of each organ and each cancer exhibit unique expression patterns despite being part of the same organ system.
Measuring host immune response status by simultaneous quantitative measurement of activity of signal transduction pathways that coordinate functional activity of immune cells from innate and adaptive immune system
Authors: Bouwman Wilbert , Verhaegh Wim , van Doorn Arie , van de Stolpe Anja 
Published: October 2021

AbstractFor many diseases, including cancer, viral infections such as COVID-19, bacterial infections, and auto-immune diseases, the immune response is a major determinant of progression, response to therapy, and clinical outcome. Innate and adaptive immune response are controlled by coordinated activity of multiple immune cell types. The functional activity state of immune cells is determined by cellular signal transduction pathways (STPs). A novel mRNA-based signaling pathway assay platform has been developed to quantitatively measure relevant STP activities in all types of immune cells and mixed immune cell samples for experimental and diagnostic purposes. We generated a STP activity profile, termed Immune-Pathway Activity Profile (I-PAP), for a variety of immune cell types in resting and activated state, and provide a first example for use in patient samples.MethodsThe technology to measure STP activity has been described for androgen and estrogen receptor, PI3K, MAPK, TGFβ, Notch, NFκB, JAK-STAT1/2, and JAK-STAT3 pathways. STP activity was measured on Affymetrix expression microarray data from preclinical studies containing public data from different types of immune cells, resting/naïve or immune-activated in vitro, to establish I-PAPs. Subsequently data from a clinical study on rheumatoid arthritis were analyzed.ResultsI-PAPs of naïve/resting and immune-activated CD4+ and CD8+ T cells, T helper cells, B cells, NK cells, monocytes, macrophages, and dendritic cells were established and in agreement with known experimental immunobiology. In whole blood samples of rheumatoid arthritis patients TGFβ pathway activity was increased; JAK-STAT3 pathway activity was selectively increased in female patients. In naïve CD4+ Tregs TGFβ pathway activity was increased, while in memory T effector cells JAK-STAT3 pathway activity tended to increase, suggesting that these immune cell types contributed to whole blood analysis results.ConclusionSTP assay technology (currently being converted to qPCR-based assays) makes it possible to directly measure functional activity of cells of the innate and adaptive immune response enabling quantitative assessment of the immune response of an individual patient. Envisioned utility lies in (1) prediction and monitoring of response to immunomodulatory treatments for a variety of immune-mediated diseases, including RA; (2) uncovering novel treatment targets; (3) improvement and standardization of in vitro immunology research and drug development.
Quantitative study of alpha-synuclein prion-like spreading in fully oriented reconstructed neural networks reveals non-synaptic dissemination of seeding aggregates
Authors: Courte Josquin , Le Ngoc Anh , Bousset Luc , Melki Ronald , Villard Catherine , Peyrin Jean-Michel 
Published: October 2021

AbstractThe trans-neuronal spread of protein aggregates in a prion-like manner underlies the progression of neuronal lesions in the brain of patients with synucleinopathies such as Parkinson’s disease. Despite being studied actively, the mechanisms of alpha-synuclein (aSyn) aggregates propagation remain poorly understood. This hinders the development of therapeutic approaches aiming at preventing the spatial progression of intracellular inclusions in neural networks. To assess the role of synaptic structures and neuron characteristics in the transfer efficiency of aggregates with seeding propensity, we developed a novel microfluidic culture system which allows for the first time to reconstruct in vitro fully oriented and synaptically connected neural networks. This is achieved by filtering axonal growth with unidirectional “axon valves” microchannels. We exposed the presynaptic compartment of reconstructed networks to well characterized human aSyn aggregates differing in size: Fibrils and Oligomers. Both aggregates were transferred to postsynaptic neurons through active axonal transport, albeit with poor efficiency. By manipulating network maturity, we compared the transfer rate of aggregates in networks with distinct levels of synaptic connectivity. Surprisingly, we found that transfer efficiency was lower in mature networks with higher synaptic connectivity. We then investigated the seeding efficiency of endogenous aSyn in the postsynaptic population. We found that exposure to Fibrils, and not Oligomers, resulted in low efficiency trans-neuronal seeding which was restricted to postsynaptic axons. Finally, we assessed the impact of neuron characteristics and aSyn expression on the propagation of aSyn aggregates. By reconstructing chimeric networks, we found that neuron characteristics, such as the brain region from which they originate or aSyn expression levels, did not significantly impact aggregates transfer, and observed no trans-neuronal seeding where the presynaptic population did not express aSyn. Overall, we demonstrate that this novel platform uniquely allows the quantitative interrogation of original aspects of the trans-neuronal propagation of seeding pathogenic entities.
Heterologous expression of Dehalobacter spp. respiratory reductive dehalogenases in Escherichia coli
Authors: Picott Katherine J. , Flick Robert , Edwards Elizabeth A. 
Published: October 2021

AbstractReductive dehalogenases (RDases) are a family of redox enzymes that are required for anaerobic organohalide respiration, a microbial process that is useful in bioremediation. Structural and mechanistic studies of these enzymes have been greatly impeded due to challenges in RDase heterologous expression, primarily because of their cobamide-dependence. There have been a few successful attempts at RDase production in unconventional heterologous hosts, but a robust method has yet to be developed. In this work we outline a novel respiratory RDase expression system using Escherichia coli as the host. The overexpression of E. coli’s cobamide transport system, btu, and RDase expression under anaerobic conditions were established to be essential for the expression of active RDases from Dehalobacter - an obligate organohalide respiring bacterium. The expression system was validated on six RDase enzymes with amino acid sequence identities ranging from >30-95%. Dehalogenation activity was verified for each RDase by assaying cell-free extracts of small-scale expression cultures on various chlorinated substrates including chloroalkanes, chloroethenes, and hexachlorocyclohexanes. Two RDases, TmrA from Dehalobacter sp. UNSWDHB and HchA from Dehalobacter sp. HCH1, were purified by nickel affinity chromatography. Incorporation of both the cobamide and iron-sulfur cluster cofactors was verified, and the specific activity of TmrA was found to be consistent with that of the native enzyme. The heterologous expression of respiratory RDases, particularly from obligate organohalide respiring bacteria, has been extremely challenging and unreliable. Here we present a relatively straightforward E. coli expression system that has performed well for a variety of Dehalobacter spp. RDases.IMPORTANCEUnderstanding microbial reductive dehalogenation is important to refine the global halogen cycle and to improve bioremediation of halogenated contaminants; however, studies of the family of enzymes responsible are limited. Characterization of reductive dehalogenase enzymes has largely eluded researchers due to the lack of a reliable and high-yielding production method. We are presenting an approach to express reductive dehalogenase enzymes from Dehalobacter, a key group of organisms used in bioremediation, in E. coli. This expression system will propel the study of reductive dehalogenases by facilitating their production and isolation, allowing researchers to pursue more in-depth questions about the activity and structure of these enzymes. This platform will also provide a starting point to improve the expression of reductive dehalogenases from many other organisms.
seRNAPAM-1regulates skeletal muscle satellite cell activation and aging throughtransregulation ofTimp2expression synergistically with Ddx5
Authors: So Karl Kam Hei , Huang Yile , Zhang Suyang , He Liangqiang , Li Yuying , Chen Xiaona , Zhao Yu , Ding Yingzhe , Zhou Jiajian , Yuan Jie , Sham Mai Har , Sun Hao , Wang Huating 
Published: October 2021

AbstractMuscle satellite cells (SCs) are responsible for muscle homeostasis and regeneration; and lncRNAs play important roles in regulating SC activities. Here in this study, we identifyPAM-1(Pax7 Associated Muscle lncRNA) that is induced in activated SCs to promote SC activation into myoblast cells upon injury.PAM-1is generated from a myoblast specific super-enhancer (SE); as a seRNA it binds with a number of target genomic loci predominantly intrans. Further studies demonstrate that it interacts with Ddx5 to tetherPAM-1SE to it inter-chromosomal targetsTimp2 and Vimto activate the gene expression. Lastly, we show thatPAM-1expression is increased in aging SCs, which leads to enhanced inter-chromosomal interaction and target genes up-regulation. Altogether, our findings identifyPAM-1as a previously unknown lncRNA that regulates both SC activation and aging through itstransgene regulatory activity.
High-throughput Activity Assay for Screening Inhibitors of the SARS-CoV-2 Mac1 Macrodomain
Authors: Dasovich Morgan , Zhuo Junlin , Goodman Jack A. , Thomas Ajit , McPherson Robert Lyle , Jayabalan Aravinth Kumar , Busa Veronica F. , Cheng Shang-Jung , Murphy Brennan A. , Redinger Karli R. , Tsukamoto Takashi , Slusher Barbara , Bosch Jürgen , Wei Huijun , Leung Anthony K. L. 
Published: October 2021

ABSTRACTMacrodomains are a class of conserved ADP-ribosylhydrolases expressed by viruses of pandemic concern, including coronaviruses and alphaviruses. Viral macrodomains are critical for replication and virus-induced pathogenesis; therefore, these enzymes are a promising target for antiviral therapy. However, no potent or selective viral macrodomain inhibitors currently exist, in part due to the lack of a high-throughput assay for this class of enzymes. Here, we developed a high-throughput ADP-ribosylhydrolase assay using the SARS-CoV-2 macrodomain Mac1. We performed a pilot screen which identified dasatinib and dihydralazine as ADP-ribosylhydrolase inhibitors. Importantly, dasatinib does not inhibit MacroD2, the closest Mac1 homolog in humans. Our study demonstrates the feasibility of identifying selective inhibitors based on ADP-ribosylhydrolase activity, paving the way for screening large compound libraries to identify improved macrodomain inhibitors and explore their potential as antiviral therapies for SARS-CoV-2 and future viral threats.
Sumo-mediated recruitment allows timely function of the Yen1 nuclease in mitotic cells
Authors: Dorison Hugo , Talhaoui Ibtissam , Mazón Gerard 
Published: October 2021

AbstractThe modification of DNA damage response proteins with Sumo is an important mechanism to orchestrate a timely and orderly recruitment of repair factors to damaged sites. After replication stress and double-strand break formation a number of repair factors are Sumoylated and interact with other Sumoylated factors, including the nuclease Yen1. Yen1 plays a critical role to ensure genome stability and unperturbed chromosome segregation by removing covalently linked DNA intermediates that are formed by homologous recombination. Here we show how this important role of Yen1 is dependent on interactions mediated by non-covalent binding to Sumoylated partners. Mutations in the motifs that allow Sumo-mediated recruitment of Yen1 impair its ability to resolve DNA intermediates and result in increased genome instability and chromosome mis-segregation.
Evolution of chlorophyll degradation is associated with plant transition to land
Authors: Schumacher Isabel , Menghini Damian , Ovinnikov Serguei , Hauenstein Mareike , Fankhauser Nick , Zipfel Cyril , Hörtensteiner Stefan , Aubry Sylvain 
Published: October 2021

AbstractColonization of land by green plants (Viridiplantae) some 500 million years ago was made possible by large metabolic and biochemical adaptations. Chlorophyll, the central pigment of photosynthesis, is highly photo-active. In order to mitigate deleterious effects of pigment accumulation, some plants have evolved a coordinated pathway to deal with chlorophyll degradation end-products, so-called phyllobilins. This pathway has been so far mostly unravelled inArabidopsis thaliana. Here, large-scale comparative phylogenomic coupled to an innovative biochemical characterization strategy of phyllobilins allow a better understanding how such a pathway appeared in Viridiplantae. Our analysis reveals a stepwise evolution of the canonical pheophorbideamonooxygenase/phyllobilin pathway. It appears to have evolved gradually, first in chlorophyte’s chloroplasts, to ensure multicellularity by detoxifying chlorophyll catabolites, and in charophytes outside chloroplasts to allow adaptation of embryophytes to land. At least six out of the eight genes involved in the pathway were already present in the last common ancestor of green plants. This strongly suggests parallel evolution of distinct enzymes catalysing similar reactions in various lineages, particularly for the dephytylation step. Together, our study suggests that chlorophyll degradation accompanied the transition from water to land, and was therefore of great importance for plant diversification.
Zika virus impacts extracellular vesicle composition and cellular gene expression in macaque early gestation trophoblasts
Authors: Block Lindsey N. , Schmidt Jenna Kropp , McKeon Megan C. , Bowman Brittany D. , Wiepz Gregory J. , Golos Thaddeus G. 
Published: October 2021

AbstractZika virus (ZIKV) infection at the maternal-placental interface is associated with adverse pregnancy outcomes including fetal demise and pregnancy loss. To determine how infection impacts placental trophoblasts, we utilized rhesus macaque trophoblast stem cells (TSC) that can be differentiated into early gestation syncytiotrophoblasts (ST) and extravillous trophoblasts (EVT). TSCs and STs, but not EVTs, were highly permissive to productive infection with ZIKV strain DAK AR 41524. The impact of ZIKV on the cellular transcriptome showed that infection of TSCs and STs increased expression of immune related genes, including those involved in type I and type III interferon responses. ZIKV exposure altered extracellular vesicle (EV) protein, mRNA, and miRNA cargo, regardless of productive infection. These findings suggest that early gestation macaque TSCs and STs are permissive to ZIKV infection, and that EV analysis may provide a foundation for identifying non-invasive biomarkers of placental infection in a highly translational model.
Characterization of divalent cation interactions with AASTY native nanodiscs
Authors: Timcenko Milena , Autzen Anton A. A. , Autzen Henriette E. 
Published: October 2021

AbstractAmphiphilic copolymers show promise in extracting membrane proteins directly from lipid bilayers into ‘native nanodiscs’. However, many such copolymers are polyanionic and sensitive to divalent cations, limiting their applicability. We characterize the Ca2+ and Mg2+ sensitivity of poly(acrylic acid-co-styrene) (AASTY) copolymers with analytical UV and fluorescent size exclusion chromatography, enabling us to separate signals from nanodiscs, copolymers, and soluble aggregates. We find that divalent cations promote aggregation and precipitation of both free and lipid bound copolymers. We see that excess, free copolymer acts as a ‘cation sink’ that protects nanodiscs from Ca2+ induced aggregation. Removal of the free copolymer through dialysis induces aggregation that can be mitigated by KCl. Finally, we find that the nanodisc size is dynamic and dependent on lipid concentration. Our results offer insight to nanodisc behaviour, and can help guide experimental design, aimed at mitigating the shortcomings inherent in negatively charged nanodisc forming copolymers.
Screening coffee genotypes for Cercospora coffeicola resistance in Brazil
Authors: Ramos Juliana Barros , Resende Mario Lucio Vilela de , Botelho Deila Magna dos Santos , Pereira Renata Cristina Martins , Reichel Tharyn , Balieiro André Augusto Ferreira , Botega Gustavo Pucci , Rezende Abrahão Juliana Costa de 
Published: October 2021

AbstractSeveral efforts have been made by many researchers worldwide to develop coffee plants resistant to different Cercospora species; however, studies concerning C. coffeicola, specifically, are still incipient. In the present study, a blend of strains from this pathogen was inoculated into 18 Brazilian commercial cultivars, a coffee clone of Arabica, as well as into 41 accessions from the Germplasm Collection of Minas Gerais, to evaluate the genetic resistance ability within the population and select superior genotypes for the breeding program. After predicting genotypic values of the evaluated material, the most efficient way to select genotypes based on the data of severity to brown eye spot (BES) was also examined. Moreover, the action of defense mechanisms against C. coffeicola attacks was investigated by assessing the levels of total soluble phenolic compounds and soluble lignin in contrasting genotypes regarding disease susceptibility. Based on the results, the accession MG 1207 Sumatra demonstrated an intrinsic genetic capacity to maintain low levels of severity to brown eye spot. This genotype can thus substantially contribute to the development of new cultivars, which may lead to reduced use of pesticides. This study also evidenced that four evaluations of severity is enough to reach accuracy and efficiency for the severity of BES, thus providing expressive genetic gains. Finally, it is suggested that the levels of lignin and phenolic compounds are not associated with the resistance of coffee genotypes to brown eye spot.