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  1. Article ; Online: Large-scale capture of hidden fluorescent labels for training generalizable markerless motion capture models

    Daniel J. Butler / Alexander P. Keim / Shantanu Ray / Eiman Azim

    Nature Communications, Vol 14, Iss 1, Pp 1-

    2023  Volume 16

    Abstract: Abstract Deep learning-based markerless tracking has revolutionized studies of animal behavior. Yet the generalizability of trained models tends to be limited, as new training data typically needs to be generated manually for each setup or visual ... ...

    Abstract Abstract Deep learning-based markerless tracking has revolutionized studies of animal behavior. Yet the generalizability of trained models tends to be limited, as new training data typically needs to be generated manually for each setup or visual environment. With each model trained from scratch, researchers track distinct landmarks and analyze the resulting kinematic data in idiosyncratic ways. Moreover, due to inherent limitations in manual annotation, only a sparse set of landmarks are typically labeled. To address these issues, we developed an approach, which we term GlowTrack, for generating orders of magnitude more training data, enabling models that generalize across experimental contexts. We describe: a) a high-throughput approach for producing hidden labels using fluorescent markers; b) a multi-camera, multi-light setup for simulating diverse visual conditions; and c) a technique for labeling many landmarks in parallel, enabling dense tracking. These advances lay a foundation for standardized behavioral pipelines and more complete scrutiny of movement.
    Keywords Science ; Q
    Subject code 006
    Language English
    Publishing date 2023-09-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: A comprehensive metagenomics framework to characterize organisms relevant for planetary protection

    David C. Danko / Maria A. Sierra / James N. Benardini / Lisa Guan / Jason M. Wood / Nitin Singh / Arman Seuylemezian / Daniel J. Butler / Krista Ryon / Katerina Kuchin / Dmitry Meleshko / Chandrima Bhattacharya / Kasthuri J. Venkateswaran / Christopher E. Mason

    Microbiome, Vol 9, Iss 1, Pp 1-

    2021  Volume 15

    Abstract: Abstract Background Clean rooms of the Space Assembly Facility (SAF) at the Jet Propulsion Laboratory (JPL) at NASA are the final step of spacecraft cleaning and assembly before launching into space. Clean rooms have stringent methods of air-filtration ... ...

    Abstract Abstract Background Clean rooms of the Space Assembly Facility (SAF) at the Jet Propulsion Laboratory (JPL) at NASA are the final step of spacecraft cleaning and assembly before launching into space. Clean rooms have stringent methods of air-filtration and cleaning to minimize microbial contamination for exoplanetary research and minimize the risk of human pathogens, but they are not sterile. Clean rooms make a selective environment for microorganisms that tolerate such cleaning methods. Previous studies have attempted to characterize the microbial cargo through sequencing and culture-dependent protocols. However, there is not a standardized metagenomic workflow nor analysis pipeline for spaceflight hardware cleanroom samples to identify microbial contamination. Additionally, current identification methods fail to characterize and profile the risk of low-abundance microorganisms. Results A comprehensive metagenomic framework to characterize microorganisms relevant for planetary protection in multiple cleanroom classifications (from ISO-5 to ISO-8.5) and sample types (surface, filters, and debris collected via vacuum devices) was developed. Fifty-one metagenomic samples from SAF clean rooms were sequenced and analyzed to identify microbes that could potentially survive spaceflight based on their microbial features and whether the microbes expressed any metabolic activity or growth. Additionally, an auxiliary testing was performed to determine the repeatability of our techniques and validate our analyses. We find evidence that JPL clean rooms carry microbes with attributes that may be problematic in space missions for their documented ability to withstand extreme conditions, such as psychrophilia and ability to form biofilms, spore-forming capacity, radiation resistance, and desiccation resistance. Samples from ISO-5 standard had lower microbial diversity than those conforming to ISO-6 or higher filters but still carried a measurable microbial load. Conclusions Although the extensive cleaning processes limit the ...
    Keywords Planetary Protection ; Spacecraft Assembly Facility ; Extremophile ; Microbial profiling ; Microbial ecology ; QR100-130
    Language English
    Publishing date 2021-04-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: A verified genomic reference sample for assessing performance of cancer panels detecting small variants of low allele frequency

    Wendell Jones / Binsheng Gong / Natalia Novoradovskaya / Dan Li / Rebecca Kusko / Todd A. Richmond / Donald J. Johann / Halil Bisgin / Sayed Mohammad Ebrahim Sahraeian / Pierre R. Bushel / Mehdi Pirooznia / Katherine Wilkins / Marco Chierici / Wenjun Bao / Lee Scott Basehore / Anne Bergstrom Lucas / Daniel Burgess / Daniel J. Butler / Simon Cawley /
    Chia-Jung Chang / Guangchun Chen / Tao Chen / Yun-Ching Chen / Daniel J. Craig / Angela del Pozo / Jonathan Foox / Margherita Francescatto / Yutao Fu / Cesare Furlanello / Kristina Giorda / Kira P. Grist / Meijian Guan / Yingyi Hao / Scott Happe / Gunjan Hariani / Nathan Haseley / Jeff Jasper / Giuseppe Jurman / David Philip Kreil / Paweł Łabaj / Kevin Lai / Jianying Li / Quan-Zhen Li / Yulong Li / Zhiguang Li / Zhichao Liu / Mario Solís López / Kelci Miclaus / Raymond Miller / Vinay K. Mittal

    Genome Biology, Vol 22, Iss 1, Pp 1-

    2021  Volume 38

    Abstract: Abstract Background Oncopanel genomic testing, which identifies important somatic variants, is increasingly common in medical practice and especially in clinical trials. Currently, there is a paucity of reliable genomic reference samples having a ... ...

    Abstract Abstract Background Oncopanel genomic testing, which identifies important somatic variants, is increasingly common in medical practice and especially in clinical trials. Currently, there is a paucity of reliable genomic reference samples having a suitably large number of pre-identified variants for properly assessing oncopanel assay analytical quality and performance. The FDA-led Sequencing and Quality Control Phase 2 (SEQC2) consortium analyze ten diverse cancer cell lines individually and their pool, termed Sample A, to develop a reference sample with suitably large numbers of coding positions with known (variant) positives and negatives for properly evaluating oncopanel analytical performance. Results In reference Sample A, we identify more than 40,000 variants down to 1% allele frequency with more than 25,000 variants having less than 20% allele frequency with 1653 variants in COSMIC-related genes. This is 5–100× more than existing commercially available samples. We also identify an unprecedented number of negative positions in coding regions, allowing statistical rigor in assessing limit-of-detection, sensitivity, and precision. Over 300 loci are randomly selected and independently verified via droplet digital PCR with 100% concordance. Agilent normal reference Sample B can be admixed with Sample A to create new samples with a similar number of known variants at much lower allele frequency than what exists in Sample A natively, including known variants having allele frequency of 0.02%, a range suitable for assessing liquid biopsy panels. Conclusion These new reference samples and their admixtures provide superior capability for performing oncopanel quality control, analytical accuracy, and validation for small to large oncopanels and liquid biopsy assays.
    Keywords Biology (General) ; QH301-705.5 ; Genetics ; QH426-470
    Subject code 519
    Language English
    Publishing date 2021-04-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Cross-oncopanel study reveals high sensitivity and accuracy with overall analytical performance depending on genomic regions

    Binsheng Gong / Dan Li / Rebecca Kusko / Natalia Novoradovskaya / Yifan Zhang / Shangzi Wang / Carlos Pabón-Peña / Zhihong Zhang / Kevin Lai / Wanshi Cai / Jennifer S. LoCoco / Eric Lader / Todd A. Richmond / Vinay K. Mittal / Liang-Chun Liu / Donald J. Johann / James C. Willey / Pierre R. Bushel / Ying Yu /
    Chang Xu / Guangchun Chen / Daniel Burgess / Simon Cawley / Kristina Giorda / Nathan Haseley / Fujun Qiu / Katherine Wilkins / Hanane Arib / Claire Attwooll / Kevin Babson / Longlong Bao / Wenjun Bao / Anne Bergstrom Lucas / Hunter Best / Ambica Bhandari / Halil Bisgin / James Blackburn / Thomas M. Blomquist / Lisa Boardman / Blake Burgher / Daniel J. Butler / Chia-Jung Chang / Alka Chaubey / Tao Chen / Marco Chierici / Christopher R. Chin / Devin Close / Jeffrey Conroy / Jessica Cooley Coleman / Daniel J. Craig

    Genome Biology, Vol 22, Iss 1, Pp 1-

    2021  Volume 23

    Abstract: Abstract Background Targeted sequencing using oncopanels requires comprehensive assessments of accuracy and detection sensitivity to ensure analytical validity. By employing reference materials characterized by the U.S. Food and Drug Administration-led ... ...

    Abstract Abstract Background Targeted sequencing using oncopanels requires comprehensive assessments of accuracy and detection sensitivity to ensure analytical validity. By employing reference materials characterized by the U.S. Food and Drug Administration-led SEquence Quality Control project phase2 (SEQC2) effort, we perform a cross-platform multi-lab evaluation of eight Pan-Cancer panels to assess best practices for oncopanel sequencing. Results All panels demonstrate high sensitivity across targeted high-confidence coding regions and variant types for the variants previously verified to have variant allele frequency (VAF) in the 5–20% range. Sensitivity is reduced by utilizing VAF thresholds due to inherent variability in VAF measurements. Enforcing a VAF threshold for reporting has a positive impact on reducing false positive calls. Importantly, the false positive rate is found to be significantly higher outside the high-confidence coding regions, resulting in lower reproducibility. Thus, region restriction and VAF thresholds lead to low relative technical variability in estimating promising biomarkers and tumor mutational burden. Conclusion This comprehensive study provides actionable guidelines for oncopanel sequencing and clear evidence that supports a simplified approach to assess the analytical performance of oncopanels. It will facilitate the rapid implementation, validation, and quality control of oncopanels in clinical use.
    Keywords Oncopanel sequencing ; Target enrichment ; Molecular diagnostics ; Reproducibility ; Analytical performance ; Precision medicine ; Biology (General) ; QH301-705.5 ; Genetics ; QH426-470
    Language English
    Publishing date 2021-04-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Host, Viral, and Environmental Transcriptome Profiles of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)

    Daniel J Butler / Christopher Mozsary / Cem Meydan / David C Danko / Jonathan Foox / Joel Rosiene / Alon Shaiber / Ebrahim Afshinnekoo / Matthew MacKay / Fritz J Sedlazeck / Nikolay A Ivanov / Maria A Sierra / Diana Pohle / Michael Zeitz / Vijendra Ramlall / Undina Gisladottir / Craig D Westover / Krista Ryon / Benjamin Young /
    Chandrima Bhattacharya / Phyllis Ruggiero / Bradley W Langhorst / Nathan A Tanner / Justyn Gawrys / Dmitry Meleshko / Dong Xu / Jenny Xiang / Angelika Iftner / Daniela Bezdan / John Sipley / Lin Cong / Arryn Craney / Priya Velu / Ari Melnick / Iman A Hajirasouliha / Thomas Iftner / Mirella Salvatore / Massimo Loda / Lars F Westblade / Shawn Levy / Melissa Cushing / Nicholas P Tatonetti / Marcin Imielinski / Hanna Rennert / Christopher Mason

    Abstract: The pandemic from the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) led to >170,000 deaths, including >14,500 in New York City (NYC) alone. This pandemic highlighted a pressing clinical and public health need for rapid, scalable ... ...

    Abstract The pandemic from the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) led to >170,000 deaths, including >14,500 in New York City (NYC) alone. This pandemic highlighted a pressing clinical and public health need for rapid, scalable diagnostics that can detect SARS-CoV-2 infection, interrogate strain evolution, and map host response in patients. To address these challenges, we designed a fast (30 minute) colorimetric test to identify SARS-CoV-2 infection and simultaneously developed a large-scale shotgun metatranscriptomic profiling platform for nasopharyngeal swabs. Both technologies were used to profile 338 clinical specimens tested for SARS-CoV-2 and 86 NYC subway samples, creating a broad molecular picture of the COVID-19 epidemic in NYC. Our results nominate a novel, NYC-enriched SARS-CoV-2 subclade, reveal specific host responses in ACE pathways, and find medication risks associated with SARS-CoV-2 infection and ACE inhibitors. Our findings have immediate applications to SARS-CoV-2 diagnostics, public health monitoring, and therapeutic development.
    Keywords covid19
    Publisher biorxiv
    Document type Article ; Online
    DOI 10.1101/2020.04.20.048066
    Database COVID19

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  6. Article ; Online: Global Genetic Cartography of Urban Metagenomes and Anti-Microbial Resistance

    David C Danko / Daniela Bezdan / Ebrahim Afshinnekoo / Sofia Ahsanuddin / Josue Alicea / Chandrima Bhattacharya / Malay Bhattacharyya / Ran Blekhman / Daniel J Butler / Eduardo Castro-Nallar / Ana M Canas / Aspassia D Chatziefthimiou / Kern Rei Chng / David A Coil / Denise Syndercombe Court / Robert W Crawford / Christelle Desnues / Emmanuel Dias-Neto / Daisy Donnellan /
    Marius Dybwad / Jonathan A Eisen / Eran Elhaik / Danilo Ercolini / Francesca De Filippis / Alina Frolova / Alexandra B Graf / David C Green / Patrick K H Lee / Jochen Hecht / Mark Hernandez / Soojin Jang / Andre Kahles / Mikhail Karasikov / Kaymisha Knights / Nikos C Kyrpides / Per Ljungdahl / Abigail Lyons / Gabriella Mason-Buck / Ken McGrath / Emmanuel F Mongodin / Harun Mustafa / Beth Mutai / Niranjan Nagarajan / Russell Y Neches / Amanda Ng / Marina Nieto-Caballero / Olga Nikolayeva / Tatyana Nikolayeva / Houtan Noushmehr / Manuela Oliveira / Stephan Ossowski / Olayinka O Osuolale / David Paez-Espino / Eileen Png / Nicolas Rascovan / Hugues Richard / Gunnar Ratsch / Jorge L Sanchez / Lynn M Schriml / Heba Shaaban / Leming Shi / Maria A Sierra / Le Huu Song / Haruo Suzuki / Dominique Thomas / Klas I Udekwu / Juan A Ugalde / Brandon Valentine / Dimitar I Vassilev / Elena Vayndorf / Marcus H Y Leung / Ben Young / Maria M Zambrano / Jifeng Zhu / Sibo Zhu / Pawel P Labaj / Christopher E Mason

    Abstract: Although studies have shown that urban environments and mass-transit systems have distinct genetic profiles, there are no systematic worldwide studies of these dense, human microbial ecosystems. To address this gap in knowledge, we created a global ... ...

    Abstract Although studies have shown that urban environments and mass-transit systems have distinct genetic profiles, there are no systematic worldwide studies of these dense, human microbial ecosystems. To address this gap in knowledge, we created a global metagenomic and antimicrobial resistance (AMR) atlas of urban mass transit systems from 60 cities, spanning 4,728 samples and 4,424 taxonomically-defined microorganisms collected for three years. This atlas provides an annotated, geospatial profile of microbial strains, functional characteristics, antimicrobial resistance markers, and novel genetic elements, including 10,928 novel predicted viral species, 1302 novel bacteria, and 2 novel archaea. Urban microbiomes often resemble human commensal microbiomes from the skin and airways, but also contain a consistent "core" of 31 species which are predominantly not human commensal species. Samples show distinct microbial signatures which may be used to accurately predict properties of their city of origin including population, proximity to the coast, and taxonomic profile. These data also show that AMR density across cities varies by several orders of magnitude, including many AMRs present on plasmids with cosmopolitan distributions. Together, these results constitute a high-resolution, global metagenomic atlas, which enables the discovery of new genetic components of the built human environment, highlights potential forensic applications, and provides an essential first draft of the global AMR burden of the world's cities.
    Keywords covid19
    Publisher biorxiv
    Document type Article ; Online
    DOI 10.1101/724526
    Database COVID19

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