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      Linking the effects of helminth infection, diet and the gut microbiota with human whole-blood signatures

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          Abstract

          Helminth infection and dietary intake can affect the intestinal microbiota, as well as the immune system. Here we analyzed the relationship between fecal microbiota and blood profiles of indigenous Malaysians, referred to locally as Orang Asli, in comparison to urban participants from the capital city of Malaysia, Kuala Lumpur. We found that helminth infections had a larger effect on gut microbial composition than did dietary intake or blood profiles. Trichuris trichiura infection intensity also had the strongest association with blood transcriptional profiles. By characterizing paired longitudinal samples collected before and after deworming treatment, we determined that changes in serum zinc and iron levels among the Orang Asli were driven by changes in helminth infection status, independent of dietary metal intake. Serum zinc and iron levels were associated with changes in the abundance of several microbial taxa. Hence, there is considerable interplay between helminths, micronutrients and the microbiota on the regulation of immune responses in humans.

          Author summary

          Parasitic intestinal worms and gut bacteria occupy the same space, but we do not understand the nature and scope of their interaction. This is further complicated by dietary effects on the gut bacteria, as well as the immune responses of the host. To better understand these complex interactions, we compared individuals living in indigenous communities in Malaysia, where worm infections are common, with people living in the capital of Malaysia, who were not infected with worms. Data collected included burden of infection, a dietary survey, clinical tests, RNA profiles on blood samples and gut bacteria composition. By collecting data before and after treating the indigenous Malaysians with deworming medication, we could determine what was associated with changes in worm burden following deworming. We found that worm infection had a larger effect on gut bacteria composition than did dietary intake or blood profiles. Worm burden also had the strongest association with blood RNA profiles. We found that zinc and iron levels in the blood were associated with changes in helminth infection status, independent of dietary metal intake. Our results suggest that there is considerable interplay between intestinal worms and gut bacteria with zinc and iron levels in infected people.

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          Most cited references43

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          Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample.

          The ongoing revolution in high-throughput sequencing continues to democratize the ability of small groups of investigators to map the microbial component of the biosphere. In particular, the coevolution of new sequencing platforms and new software tools allows data acquisition and analysis on an unprecedented scale. Here we report the next stage in this coevolutionary arms race, using the Illumina GAIIx platform to sequence a diverse array of 25 environmental samples and three known "mock communities" at a depth averaging 3.1 million reads per sample. We demonstrate excellent consistency in taxonomic recovery and recapture diversity patterns that were previously reported on the basis of metaanalysis of many studies from the literature (notably, the saline/nonsaline split in environmental samples and the split between host-associated and free-living communities). We also demonstrate that 2,000 Illumina single-end reads are sufficient to recapture the same relationships among samples that we observe with the full dataset. The results thus open up the possibility of conducting large-scale studies analyzing thousands of samples simultaneously to survey microbial communities at an unprecedented spatial and temporal resolution.
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            Alternative projections of mortality and disability by cause 1990–2020: Global Burden of Disease Study

            The Lancet, 349(9064), 1498-1504
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              mixOmics: An R package for ‘omics feature selection and multiple data integration

              The advent of high throughput technologies has led to a wealth of publicly available ‘omics data coming from different sources, such as transcriptomics, proteomics, metabolomics. Combining such large-scale biological data sets can lead to the discovery of important biological insights, provided that relevant information can be extracted in a holistic manner. Current statistical approaches have been focusing on identifying small subsets of molecules (a ‘molecular signature’) to explain or predict biological conditions, but mainly for a single type of ‘omics. In addition, commonly used methods are univariate and consider each biological feature independently. We introduce mixOmics, an R package dedicated to the multivariate analysis of biological data sets with a specific focus on data exploration, dimension reduction and visualisation. By adopting a systems biology approach, the toolkit provides a wide range of methods that statistically integrate several data sets at once to probe relationships between heterogeneous ‘omics data sets. Our recent methods extend Projection to Latent Structure (PLS) models for discriminant analysis, for data integration across multiple ‘omics data or across independent studies, and for the identification of molecular signatures. We illustrate our latest mixOmics integrative frameworks for the multivariate analyses of ‘omics data available from the package.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: MethodologyRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: Formal analysisRole: Visualization
                Role: Investigation
                Role: Funding acquisition
                Role: Investigation
                Role: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: 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
                16 December 2019
                December 2019
                : 15
                : 12
                : e1008066
                Affiliations
                [1 ] Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
                [2 ] Centre of Excellence for Research in AIDS (CERiA), University of Malaya, Kuala Lumpur, Malaysia
                [3 ] Department of Microbiology, New York University School of Medicine, New York, New York, United States of America
                [4 ] Department of Obstetrics and Gynecology, Faculty of Medicine, University of Malaya Medical Centre, Kuala Lumpur, Malaysia
                [5 ] Department of Paediatrics, Faculty of Medicine, University of Malaya Medical Centre, Kuala Lumpur, Malaysia
                [6 ] Department of Medicine, Division of Gastroenterology, New York University School of Medicine, New York, New York, United States of America
                [7 ] Department of Social and Preventive Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
                [8 ] University of Malaya Centre for Data Analytics, University of Malaya, Kuala Lumpur, Malaysia
                [9 ] Institute of Mathematical Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
                Washington University School of Medicine, UNITED STATES
                Author notes

                The authors have declared that no competing interests exist.

                [¤]

                Current address: Department of Pathology & Immunology, Washington University School of Medicine in St. Louis, St. Louis, Missouri, United States of America

                Author information
                http://orcid.org/0000-0002-4476-9415
                http://orcid.org/0000-0002-1315-9489
                http://orcid.org/0000-0002-7308-8707
                http://orcid.org/0000-0002-0306-4419
                http://orcid.org/0000-0003-4433-9738
                Article
                PPATHOGENS-D-19-00594
                10.1371/journal.ppat.1008066
                6913942
                31841569
                62cf7fc7-6883-41af-ad89-45055ed081db
                © 2019 Lee et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 28 March 2019
                : 3 September 2019
                Page count
                Figures: 8, Tables: 0, Pages: 30
                Funding
                Funded by: Ministry of Higher Education Malaysia under Fundamental Research Grant Scheme
                Award ID: grant FP004-2017A
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100004386, Universiti Malaya;
                Award ID: BKS005-2017
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000060, National Institute of Allergy and Infectious Diseases;
                Award ID: AI093811
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000060, National Institute of Allergy and Infectious Diseases;
                Award ID: AI094166
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000062, National Institute of Diabetes and Digestive and Kidney Diseases;
                Award ID: DK103788
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000862, Doris Duke Charitable Foundation;
                Award Recipient :
                NYU Langone's Genome Technology Center (GTC) is part of an institution-shared resource partially supported by the Cancer Center (support grant P30CA016087) at the Laura and Isaac Perlmutter Cancer Center. We also thank the NYU IT High Performance Computing resources, services, and staff expertise. This work was supported by the NIH NIAID (grants AI093811 and AI094166 to P.L.), NIDDK (grant DK103788 to P.L.), Vilcek Foundation (M.S.T.), AAI (M.S.T.), University of Malaya/Ministry of Higher Education Malaysia High Impact Research (grant UM.C/625/HIR/MOE/MED/23 to Y.A.L.L. and P.L), University of Malaya (grant BKS005-2017 to Y.A.L.L.), Ministry of Higher Education Malaysia under Fundamental Research Grant Scheme (grant FP004-2017A to Y.A.L.L. and S.C.L.), Doris Duke (I.C.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Body Fluids
                Blood
                Medicine and Health Sciences
                Anatomy
                Body Fluids
                Blood
                Biology and Life Sciences
                Physiology
                Body Fluids
                Blood
                Medicine and Health Sciences
                Physiology
                Body Fluids
                Blood
                Medicine and Health Sciences
                Parasitic Diseases
                Helminth Infections
                Physical Sciences
                Chemistry
                Chemical Elements
                Zinc
                Biology and Life Sciences
                Genetics
                Gene Expression
                Gene Regulation
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbiome
                Biology and Life Sciences
                Genetics
                Genomics
                Microbial Genomics
                Microbiome
                Biology and Life Sciences
                Microbiology
                Microbial Genomics
                Microbiome
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Invertebrates
                Helminths
                Biology and Life Sciences
                Organisms
                Bacteria
                Gut Bacteria
                Research and Analysis Methods
                Mathematical and Statistical Techniques
                Statistical Methods
                Multivariate Analysis
                Principal Component Analysis
                Physical Sciences
                Mathematics
                Statistics
                Statistical Methods
                Multivariate Analysis
                Principal Component Analysis
                Custom metadata
                Raw data of microbial 16S rRNA sequencing has been deposited on the European Nucleotide Archive with accesion numbers PRJEB34956 (ERP117943) and PRJEB34957 (ERP117944). Raw data of RNA-Seq has been deposited on NCBI’s Gene Expression Omnibus with accession GSE137338.

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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