bluuebunny/crossref_metadata_2025_split huggingface dataset:
COPILOT: a Containerised wOrkflow for Processing ILlumina genOtyping daTa
Authors: Patel Hamel , Lee Sang-hyuck , Breen Gerome , Menzel Stephen , Ojewunmi Oyesola , Dobson Richard J.B 
Published: July 2021

AbstractBackgroundThe Illumina genotyping microarrays generate data in image format, which is processed by the platform-specific software GenomeStudio, followed by an array of complex bioinformatics analyses. This process can be time-consuming, lead to reproducibility errors, and be a daunting task for novice bioinformaticians.ResultsHere we introduce the COPILOT (Containerised wOrkflow for Processing ILlumina genOtyping daTa) protocol, which provides an in-depth and clear guide to process raw Illumina genotype data in GenomeStudio, followed by a containerised workflow to automate an array of complex bioinformatics analyses involved in a GWAS quality control (QC). The COPILOT protocol was applied to two independent cohorts consisting of 2791 and 479 samples genotyped on the Infinium Global Screening (GSA) array with Multi-disease (MD) drop-in (~750,000 markers) and the Infinium H3Africa consortium array (~2,200,000 markers) respectively. Following the COPILOT protocol, an average sample quality improvement of 1.24% was observed across sample call rates, with notable improvement for low-quality samples. For example, from the 3270 samples processed, 141 samples had an initial sample call rate below 98%, averaging 96.6% (95% CI 95.6-97.7%), which is considered below the acceptable sample call rate threshold for a typical GWAS analysis. However, following the COPILOT protocol, all 141 samples had a call rate above 98% after QC and averaged 99.6% (95% CI 99.5-99.7%). In addition, the COPILOT pipeline automatically identified potential data issues, including gender discrepancies, heterozygosity outliers, related individuals, and population outliers through ancestry estimation.ConclusionsThe COPILOT protocol makes processing Illumina genotyping data transparent, effortless and reproducible. The container is deployable on multiple platforms, improves data quality, and the end product is analysis-ready PLINK formatted data, with a comprehensive and interactive summary report to guide the user for further data analyses.
Using topic modeling to detect cellular crosstalk in scRNA-seq
Authors: Pancheva Alexandrina , Wheadon Helen , Rogers Simon , Otto Thomas 
Published: July 2021

AbstractCell-cell interactions are vital for numerous biological processes including development, differentiation, and response to inflammation. Currently most methods for studying interactions on scRNA-seq level are based on curated databases of ligands and receptors. While those methods are useful, they are limited to our current biological knowledge. Recent advances in single cell protocols have allowed for physically interacting cells to be captured and as such we have the potential to study interactions in a complimentary way without relying on prior knowledge. We introduce a new method for detecting genes that change as a result of interaction based on Latent Dirichlet Allocation (LDA). We apply our method to synthetic datasets to demonstrate its ability to detect genes that change in an interacting population compared to a reference population. Next, we apply our approach to two datasets of physically interacting cells and identify genes that change as a result of interaction, examples include adhesion and co-stimulatory molecules which confirm physical interaction between cells. For each dataset we produce a ranking of genes that are changing in subpopulations of the interacting cells. In addition to genes, discussed in the original publications we highlight further candidates for interaction in the top 100 and 300 ranked genes. Lastly, we apply our method to a dataset generated by a standard droplet based protocol, not designed to capture interacting cells and discuss its suitability for analysing interactions. We present a method that streamlines the detection of interactions and does not require prior clustering and generation of synthetic reference profiles to detect changes in expression.Author summaryWhile scRNA-seq research is a dynamic area, progress is lacking when it comes to developing methods that allow analysis of interaction that is independent of curated resources of known interacting pairs. Recent advances of sequencing protocols have allowed for interacting cells to be captured. We propose a novel method based on LDA that captures changes in gene expression as a result of interaction. Our method does not require prior information in the form of clustering or generation of synthetic reference profiles. We demonstrate the suitability of our approach by applying it to synthetic and real datasets and manage to capture biologically interesting candidates of interaction.
Profound structural conservation of chemically cross-linked HIV-1 envelope glycoprotein experimental vaccine antigens
Authors: Martin Gregory , Russell Rebecca A , Mundsperger Philip , Harris Scarlett , Jovanoska Lu , Trajano Luiza Farache , Schiffner Torben , Fabian Katalin , Tolazzi Monica , Scarlatti Gabriella , McFarlane Leon , Cheeseman Hannah , Aldon Yoann , Breemen Marielle , Sliepen Kwinten , Katinger Dietmar , Kunert Renate , Sanders Rogier W , Shattock Robin , Ward Andrew B , Sattentau Quentin J 
Published: July 2021

AbstractChemical cross-linking is used to stabilise protein structure with additional benefits of pathogen and toxin inactivation for vaccine use, but its use is restricted by potential induction of local or global structural distortion. This is of particular importance when the protein in question requires a high degree of structural conservation for the purposes of understanding function, or for inducing a biological outcome such as elicitation of antibodies to conformationally-sensitive epitopes. The HIV-1 envelope glycoprotein (Env) trimer is metastable and shifts between different conformational states, complicating its functional analysis and use as a vaccine antigen. Here we have used the hetero-bifunctional zero-length reagent EDC to cross-link two soluble Env trimers, selected well-folded trimers using an antibody affinity column, and transferred this process to good manufacturing practice (GMP) for clinical trial use. Cross-linking enhanced GMP trimer stability to biophysical and enzyme attack, and had broadly beneficial effects on morphology, antigenicity and immunogenicity. Cryo-EM analysis revealed that cross-linking essentially completely retained overall structure with RMSDs between unmodified and cross-linked Env trimers of 0.4-0.5 Å. Despite this negligible distortion of global trimer structure we identified individual inter-subunit, intra-subunit and intra-protomer cross-links. Thus, EDC cross-linking maintains protein folding, improves stability, and is readily transferred to GMP, consistent with use of this approach in probing protein structure/function relationships and in the design of vaccines.
A multispecies amplicon sequencing approach for genetic diversity assessment in grassland plant species
Authors: Loera-Sánchez Miguel , Studer Bruno , Kölliker Roland 
Published: July 2021

AbstractGrasslands are widespread and economically relevant ecosystems at the basis of sustainable roughage production. Plant genetic diversity (PGD; i.e., within-species diversity) is related to many beneficial effects to the ecosystem functioning of grasslands. The monitoring of PGD in temperate grasslands is complicated by the multiplicity of species present and by a shortage of methods for large-scale assessment. However, the continuous advancement of high-throughput DNA sequencing approaches have improved the prospects of broad, multispecies PGD monitoring. Among them, amplicon sequencing stands out as a robust and cost-effective method.Here we report a set of twelve multispecies primer pairs that can be used for high-throughput PGD assessment in multiple grassland plant species. The loci targeted by the amplicons were selected and tested in two phases: a “discovery phase” based on a sequence capture assay (611 target nuclear loci assessed in 16 grassland plant species), which resulted in the selection of eleven loci; and a “validation phase”, in which the selected loci were targeted and sequenced using twelve multispecies primers in test populations of Dactylis glomerata L., Lolium perenne L., Festuca pratensis Huds., Trifolium pratense L. and T. repens L. The resulting multispecies amplicons had overall nucleotide diversities per species ranging from 5.19 × 10−3 to 1.29 × 10−2, which is in the range of flowering-related genes but slightly lower than pathogen resistance genes. We conclude that the methodology, the DNA sequence resources, and the amplicon-specific primer pairs reported in this study provide the basis for large-scale, multispecies PGD monitoring in grassland plants.
PEGR: a flexible management platform for reproducible epigenomic and genomic research
Authors: Shao Danying , Kellogg Gretta D. , Nematbakhsh Ali , Kuntala Prashant , Mahony Shaun , Pugh B. Franklin , Lai William KM 
Published: July 2021

AbstractReproducibility is a significant challenge in (epi)genomic research due to the complexity of experiments composed of traditional biochemistry and informatics. Recent advances have exacerbated this challenge as high-throughput sequencing data is generated at an unprecedented pace. Here we report on our development of a Platform for Epi-Genomic Research (PEGR), a web-based project management platform that tracks and quality controls experiments from conception to publication-ready figures, compatible with multiple assays and bioinformatic pipelines. It supports rigor and reproducibility for biochemists working at the wet bench, while continuing to fully support reproducibility and reliability for bioinformaticians through integration with the Galaxy platform.
Gametophyte genome activation occurs at pollen mitosis I in maize
Authors: Nelms Brad , Walbot Virginia 
Published: July 2021

AbstractFlowering plants alternate between multicellular haploid (gametophyte) and diploid (sporophyte) generations. One consequence of this life cycle is that plants face substantial selection during the haploid phase1–3. Pollen actively transcribes its haploid genome4, providing phenotypic diversity even among pollen grains from a single plant. Currently, the timing that pollen precursors first establish this independence is unclear. Starting with an endowment of transcripts from the diploid parent, when do haploid cells generated by meiosis begin to express genes? Here, we follow the shift to haploid expression in maize pollen using allele-specific RNA-sequencing (RNA-Seq) of single pollen precursors. We observe widespread biallelic expression for 11 days after meiosis, indicating that transcripts synthesized by the diploid sporophyte persist long into the haploid phase. Subsequently, there was a rapid and global conversion to monoallelic expression at pollen mitosis I (PMI), driven by active new transcription from the haploid genome. Genes expressed during the haploid phase showed reduced rates of nonsynonymous relative to synonymous substitutions (dn/ds) if they were expressed after PMI, but not before, consistent with purifying selection acting on the haploid gametophyte. This work establishes the timing with which haploid selection may act in pollen and provides a detailed time-course of gene expression during pollen development.
MYC hyperactivates WNT signaling in APC/CTNNB1-mutated colorectal cancer cells through miR-92a-dependent repression of DKK3
Authors: Sehgal Priyanka , Lanauze Claudia , Wang Xin , Hayer Katharina E. , Torres-Diz Manuel , Sela Yogev , Stanger Ben Z. , Lengner Christopher J. , Thomas-Tikhonenko Andrei 
Published: July 2021

ABSTRACTActivation of Wnt signaling is among the earliest events of colon cancer development. It is achieved either via activating mutations in the CTNNB1 gene encoding β-catenin, the key transcription factor in the Wnt pathway, or most commonly by inactivating mutations in APC, a major β-catenin binding partner and negative regulator. However, our analysis of recent Pan Cancer Atlas data revealed that CTNNB1 mutations significantly co-occur with those affecting Wnt receptor complex components (e.g., Frizzled and LRP6), underscoring the importance of additional regulatory events even in the presence of common APC/CTNNB1 mutations. In our effort to identify non-mutational hyperactivating events, we determined that KRAS-transformed murine colonocytes overexpressing direct β-catenin target MYC show significant upregulation of the Wnt signaling pathway and reduced expression of Dickkopf 3 (DKK3), a reported ligand for Wnt co-receptors. We demonstrate that Myc suppresses Dkk3 transcription through one of mir-17-92 cluster microRNAs, miR-92a. We further examined the role of DKK3 by overexpression and knockdown and discovered that DKK3 suppresses Wnt signaling in APC-null murine colonic organoids and human colon cancer cells despite the presence of downstream activating mutations in the Wnt pathway. Conversely, MYC overexpression in the same cell lines resulted in hyperactive Wnt signaling, acquisition of epithelial-to-mesenchymal transition markers, and enhanced migration and invasion and metastasis in syngeneic orthotopic mouse colon cancer model. Our results suggest that the MYC->miR-92a-|DKK3 axis hyperactivates Wnt signaling, forming a feedforward oncogenic loop.SIGNIFICANCECommon APC and CTNNB1 mutations activate Wnt signaling in colorectal cancers. Here we demonstrate that further potentiation of this pathway involves microRNA-dependent repression of the DKK3 gene by the Myc oncoprotein.
Soil fauna may buffer the negative effects of drought on alien plant invasion
Authors: Jin Huifei , Chang Liang , van Kleunen Mark , Liu Yanjie 
Published: July 2021

AbstractAlthough many studies have tested the direct effects of drought on alien plant invasion, less is known about how drought affects alien plant invasion indirectly via other groups of organisms such as soil fauna. To test for such indirect effects, we grew single plant of nine naturalized alien target species in pot-mesocosms with a native community of five native grassland species under four combinations of two drought (well-watered vs drought) and two soil-fauna-inoculation (with vs without) treatments. We found that drought decreased the absolute and the relative biomass production of the alien plants, and thus reduced their competitive advantage in the native community. Drought decreased the abundance of soil fauna, particularly the soil mites, but did not affect abundance and richness of soil herbivores. Soil-fauna inoculation increased the biomass of the native plant community and thereby decreased the relative biomass production of the alien species. The increased invasion resistance due to soil fauna, however, tended (p = 0.09) to be stronger for plants growing under well-watered conditions than under drought. Our multispecies experiment thus shows that soil fauna might help native resident communities to resist alien plant invasions, but that this effect might be diminished by drought.
Representational learning of brain responses in executive function and higher-order cognition using deep graph convolutions
Authors: Zhang Yu , Farrugia Nicolas , Dagher Alain , Bellec Pierre 
Published: July 2021

AbstractBrain decoding aims to infer human cognition from recordings of neural activity using modern neuroimaging techniques. Studies so far often concentrated on a limited number of cognitive states and aimed to classifying patterns of brain activity within a local area. This procedure demonstrated a great success on classifying motor and sensory processes but showed limited power over higher cognitive functions. In this work, we investigate a high-order graph convolution model, named ChebNet, to model the segregation and integration organizational principles in neural dynamics, and to decode brain activity across a large number of cognitive domains. By leveraging our prior knowledge on brain organization using a graph-based model, ChebNet graph convolution learns a new representation from task-evoked neural activity, which demonstrates a highly predictive signature of cognitive states and task performance. Our results reveal that between-network integration significantly boosts the decoding of high-order cognition such as visual working memory tasks, while the segregation of localized brain activity is sufficient to classify motor and sensory processes. Using twin and family data from the Human Connectome Project (n = 1,070), we provide evidence that individual variability in the graph representations of working-memory tasks are under genetic control and strongly associated with participants in-scanner behaviors. These findings uncover the essential role of functional integration in brain decoding, especially when decoding high-order cognition other than sensory and motor functions.TeaserModelling functional integration through graph convolution is a necessary step towards decoding high-order human cognition.Significance statementOver the past two decades, many studies have applied multivariate pattern analysis to decode what task a human participant is performing, based on a scan of her brain. The vast majority of these studies have however concentrated on select regions and a specific domain, because of the computational complexity of handling full brain data in a multivariate model. With the fast progress in the field of deep learning, it is now possible to decode a variety of cognitive domains simultaneously using a full-brain model. By leveraging our prior knowledge on brain organization using a graph-based model, we uncovered different organizational principles in brain decoding for motor execution and high-order cognition by modelling functional integration through graph convolution.
Histone Variant macroH2A1 Spatially and Functionally Organizes Human Papillomavirus Replication Foci in the Productive Stage of Infection
Authors: Khurana Simran , Markowitz Tovah E. , Kabat Juraj , McBride Alison A. 
Published: July 2021

ABSTRACTThe life cycle of HPV depends on keratinocyte differentiation as the virus modulates and takes advantage of cellular pathways to replicate its genome and assemble viral particles in differentiated cells. Viral genomes are amplified in nuclear replication foci in differentiated keratinocytes, and DNA repair factors from the DNA damage response signaling pathway are recruited to replicate viral DNA. The HPV genome is associated with cellular histones at all stages of the infectious cycle, and here we show the histone variant macroH2A1 is bound to the HPV genome and enriched in viral replication foci in differentiated cells. MacroH2A1 isoforms play important roles in cellular transcriptional repression, double strand break repair, and replication stress. The viral E8^E2 protein also binds to the HPV genome and inhibits viral replication and gene expression by recruiting NCoR/SMRT complexes. We show here that E8^E2 and SMRT also localize within replication foci, though not through direct interaction with macroH2A1. Conversely, transcription complexes containing RNA polymerase II and Brd4 are located on the surface of the foci. Foci generated with an HPV16 E8^E2 mutant genome are not enriched for SMRT or macroH2A1 but contain transcriptional complexes throughout the foci. We demonstrate that macroH2A1 promotes viral late transcription and propose that it does so by spatially separating replication and transcription activities within replication foci.IMPORTANCEHuman papillomaviruses are small DNA viruses that cause chronic infection of cutaneous and mucosal epithelium. In some cases, persistent infection with HPV can result in cancer, and 5% human cancers are the result of HPV infection. In differentiated cells, HPV amplifies viral DNA in nuclear replication factories and transcribes late mRNAs to produce capsid proteins. However, very little is known about the spatial organization of these activities in the nucleus. Here we show that repressive viral and cellular factors localize within the foci to supress viral transcription, while active transcription takes place on the surface. The cellular histone variant macroH2A1 is important for this spatial organization.
Nucleus size and its effect on the chromatin structure in living cells
Authors: Efremov Artem K. , Hovan Ladislav , Yan Jie 
Published: July 2021

DNA-architectural proteins play a major role in organization of chromosomal DNA in living cells by packaging it into chromatin, whose spatial conformation is determined by an intricate interplay between the DNA-binding properties of architectural proteins and physical constraints applied to the DNA by a tight nuclear space. Yet, the exact effects of the cell nucleus size on DNA-protein interactions and chromatin structure currently remain obscure. Furthermore, there is even no clear understanding of molecular mechanisms responsible for the nucleus size regulation in living cells. To find answers to these questions, we developed a general theoretical framework based on a combination of polymer field theory and transfer-matrix calculations, which showed that the nucleus size is mainly determined by the difference between the surface tensions of the nuclear envelope and the endoplasmic reticulum membrane as well as the osmotic pressure exerted by cytosolic macromolecules on the nucleus. In addition, the model demonstrated that the cell nucleus functions as a piezoelectric element, changing its electrostatic potential in a size-dependent manner. This effect has been found to have a profound impact on stability of nucleosomes, revealing a previously unknown link between the nucleus size and chromatin structure. Overall, our study provides new insights into the molecular mechanisms responsible for regulation of the cell nucleus size, as well as the potential role of nuclear organization in shaping the cell response to environmental cues.SIGNIFICANCE STATEMENTThe cell nucleus plays a central role in the life of eukaryotic cells, providing the highest level of control of intracellular processes. Depending on the stage of the cell cycle and / or surrounding environment, the size of the cell nucleus may undergo changes that are believed to cause chromatin reorganization, affecting gene transcription. At present, however, there is no clear understanding of the molecular mechanisms that may be responsible for such regulation, whose exact effect on chromatin structure remains unclear. In this study, by developing an advanced computational approach, we explore these issues from a physical perspective, revealing previously unknown mechanisms contributing to organization of the cell nucleus and chromatin.