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      Signatures of oral microbiome in HIV-infected individuals with oral Kaposi's sarcoma and cell-associated KSHV DNA

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          Abstract

          Infection by Kaposi’s sarcoma-associated herpesvirus (KSHV) is necessary for the development of Kaposi’s sarcoma (KS), which most often develops in HIV-infected individuals. KS frequently has oral manifestations and KSHV DNA can be detected in oral cells. Numerous types of cancer are associated with the alteration of microbiome including bacteria and virus. We hypothesize that oral bacterial microbiota affects or is affected by oral KS and the presence of oral cell-associated KSHV DNA. In this study, oral and blood specimens were collected from a cohort of HIV/KSHV-coinfected individuals all previously diagnosed with KS, and were classified as having oral KS with any oral cell-associated KSHV DNA status (O-KS, n = 9), no oral KS but with oral cell-associated KSHV DNA (O-KSHV, n = 10), or with neither oral KS nor oral cell-associated KSHV DNA (No KSHV, n = 10). We sequenced the hypervariable V1-V2 region of the 16S rRNA gene present in oral cell-associated DNA by next generation sequencing. The diversity, richness, relative abundance of operational taxonomic units (OTUs) and taxonomic composition of oral microbiota were analyzed and compared across the 3 studied groups. We found impoverishment of oral microbial diversity and enrichment of specific microbiota in O-KS individuals compared to O-KSHV or No KSHV individuals. These results suggest that HIV/KSHV coinfection and oral microbiota might impact one another and influence the development of oral KS.

          Author summary

          Kaposi’s sarcoma (KS) is the most common cancer occurring in HIV-infected individuals worldwide, and often involves the mouth. While infection by Kaposi’s sarcoma-associated herpesvirus (KSHV) is necessary for the development of KS, other cofactors remain unclear. In this study, we evaluated the impact of oral bacterial microbiota on the development of oral KS and the presence of oral cell-associated KSHV DNA by studying a cohort of HIV/KSHV-coinfected individuals all previously diagnosed with KS, classified as having oral KS with any oral cell-associated KSHV DNA status (O-KS), no oral KS but with oral cell-associated KSHV DNA (O-KSHV), or with neither oral KS nor oral cell-associated KSHV DNA (No KSHV). We observed impoverishment of oral microbial diversity and enrichment of specific types of microbes in O-KS individuals compared to O-KSHV or No KSHV individuals. Hence, HIV/KSHV coinfection and oral microbiota might impact one another and influence the development of oral KS.

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          FQC Dashboard: integrates FastQC results into a web-based, interactive, and extensible FASTQ quality control tool

          Abstract Summary FQC is software that facilitates quality control of FASTQ files by carrying out a QC protocol using FastQC, parsing results, and aggregating quality metrics into an interactive dashboard designed to richly summarize individual sequencing runs. The dashboard groups samples in dropdowns for navigation among the data sets, utilizes human-readable configuration files to manipulate the pages and tabs, and is extensible with CSV data. Availability and implementation FQC is implemented in Python 3 and Javascript, and is maintained under an MIT license. Documentation and source code is available at: https://github.com/pnnl/fqc. Contact joseph.brown@pnnl.gov
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            Bacterial diversity in the oral cavity of 10 healthy individuals.

            The composition of the oral microbiota from 10 individuals with healthy oral tissues was determined using culture-independent techniques. From each individual, 26 specimens, each from different oral sites at a single point in time, were collected and pooled. An 11th pool was constructed using portions of the subgingival specimens from all 10 individuals. The 16S ribosomal RNA gene was amplified using broad-range bacterial primers, and clone libraries from the individual and subgingival pools were constructed. From a total of 11,368 high-quality, nonchimeric, near full-length sequences, 247 species-level phylotypes (using a 99% sequence identity threshold) and 9 bacterial phyla were identified. At least 15 bacterial genera were conserved among all 10 individuals, with significant interindividual differences at the species and strain level. Comparisons of these oral bacterial sequences with near full-length sequences found previously in the large intestines and feces of other healthy individuals suggest that the mouth and intestinal tract harbor distinct sets of bacteria. Co-occurrence analysis showed significant segregation of taxa when community membership was examined at the level of genus, but not at the level of species, suggesting that ecologically significant, competitive interactions are more apparent at a broader taxonomic level than species. This study is one of the more comprehensive, high-resolution analyses of bacterial diversity within the healthy human mouth to date, and highlights the value of tools from macroecology for enhancing our understanding of bacterial ecology in human health.
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              Insights into the human oral microbiome

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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Funding acquisitionRole: InvestigationRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: InvestigationRole: SupervisionRole: Writing – review & editing
                Role: Funding acquisitionRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SoftwareRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS Pathog
                PLoS Pathog
                plos
                plospath
                PLoS Pathogens
                Public Library of Science (San Francisco, CA USA )
                1553-7366
                1553-7374
                17 January 2020
                January 2020
                : 16
                : 1
                : e1008114
                Affiliations
                [1 ] Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United States of America
                [2 ] Department of Electrical and Computer Engineering, University of Texas at San Antonio, San Antonio, Texas, United States of America
                [3 ] Viral Oncology Section, AIDS and Cancer Virus Program, Leidos Biomedical Research Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America
                [4 ] HIV and AIDS Malignancy Branch, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, United States of America
                [5 ] Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America
                [6 ] Department of Epidemiology and Biostatistics, The University of Texas Health San Antonio, San Antonio, Texas, United States of America
                [7 ] UPMC Hillman Cancer Center, Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America
                University of Pennsylvania, UNITED STATES
                Author notes

                I have read the journal’s policy and the authors of this manuscript have the following competing interests: TSU and RY are co-inventors on a patent related to the treatment of KSHV-associated diseases with immune modulatory compounds, and the spouse of RY is a co-inventor on a patent related to the measurement of KSHV vIL-6. These inventions were all made as part of their duties as employees of the U.S. Government, and the patents are or will be assigned to U.S. Department of Health and Human Services. The government may convey a portion of the royalties it receives from licensure of its patents to its employee inventors. RY and TSU have a CRADA with Celgene Corp. and recently conducted clinical research using drugs supplied to the NCI by Merck and Co., Hoffman LaRoche, and Bayer Healthcare. TSU receives additional research funding from Roche. None of the other co-authors have conflicts of interest to disclose.

                [¤]

                Current address: Center for Novel Cancer Therapies, Monter Cancer Center, Lake Success, New York, United States of America

                Author information
                http://orcid.org/0000-0001-6194-1742
                Article
                PPATHOGENS-D-19-01331
                10.1371/journal.ppat.1008114
                6992226
                31951641
                26b8f9d7-ae18-47a7-b2d6-3a0fba0f7937

                This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 22 July 2019
                : 27 September 2019
                Page count
                Figures: 5, Tables: 1, Pages: 18
                Funding
                Funded by: National Cancer Institute (US)
                Award ID: CA096512
                Award Recipient :
                Funded by: NCI
                Award ID: CA124332
                Award Recipient :
                Funded by: NCI
                Award ID: CA132637
                Award Recipient :
                Funded by: NCI
                Award ID: CA177377
                Award Recipient :
                Funded by: NCI
                Award ID: CA213275
                Award Recipient :
                Funded by: NCI
                Award ID: CA197153
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000072, National Institute of Dental and Craniofacial Research;
                Award ID: DE025465
                Award Recipient :
                Funded by: NCI
                Award ID: HHSN261200800001E
                Award Recipient :
                Funded by: NCI
                Award ID: Intramural Research Program
                Award Recipient :
                This work was in part supported by grants from NIH (DE025465, CA096512, CA124332, CA132637, CA177377, CA213275 and CA197153) to S-J Gao, and in part by the Intramural Research Program of the National Cancer Institute, NIH and by US federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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