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      Major antigen and paramyosin proteins as candidate biomarkers for serodiagnosis of canine infection by zoonotic Onchocerca lupi

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          Abstract

          Onchocerca lupi (Spirurida: Onchocercidae) is a filarial worm parasitizing domestic carnivores and humans. Adult nematodes usually localize beneath in the sclera or in the ocular retrobulbar of infected animals, whilst microfilariae are found in the skin. Therefore, diagnosis of O. lupi is achieved by microscopic and/or molecular detection of microfilariae from skin biopsy and/or surgical removal of adults from ocular tissues of infected hosts. An urgent non-invasive diagnostic tool for the diagnosis of O. lupi in dog is mandatory. In this study, an immunoproteomic analyses was performed using a combination of immunoblotting and mass spectrometry techniques. Onchocerca lupi major antigen ( Ol-MJA) and paramyosin ( Ol-PARA) proteins were identified as potential biomarkers for serodiagnosis. Linear epitopes were herein scanned for both proteins using high-density peptide microarray. Sera collected from dog infected with O. lupi and healthy animal controls led to the identification of 11 immunodominant antigenic peptides (n = 7 for Ol-MJA; n = 4 for Ol-PARA). These peptides were validated using sera of dogs uniquely infected with the most important filarioids infesting dogs either zoonotic ( Dirofilaria repens, Dirofilaria immitis) or not ( Acanthocheilonema reconditum and Cercopithifilaria bainae). Overall, six antigenic peptides, three for Ol-MJA and for Ol-PARA, respectively, were selected as potential antigens for the serological detection of canine O. lupi infection. The molecular and proteomic dataset herein reported should provide a useful resource for studies on O. lupi toward supporting the development of new interventions (drugs, vaccines and diagnostics) against canine onchocercosis.

          Author summary

          The diagnosis of Onchocerca lupi (Spirurida: Onchocercidae), a zoonotic nematode of domestic animals, is currently based on microscopic examination of skin snip sediments and on the identification of adults embedded in ocular nodules or by molecular assays. An urgent non-invasive diagnostic tool for the diagnosis of O. lupi in dogs is mandatory. In this context, combined immunoblotting and mass spectrometry-based analyses have been performed to identify two proteins, major antigen and paramyosin, of O. lupi. Peptides herein identified represent suitable candidate biomarker for the development of a specific diagnostic test for canine onchocercosis. An accurate, minimally invasive diagnostic method could prove useful for the control of the canine diseases, for establishing large sero-surveys, for mapping the distribution of the infection in endemic areas as well as in areas where information on the disease is not available and for the reduction of risks for human infection.

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          A new approach to rapid sequence comparison, basic local alignment search tool (BLAST), directly approximates alignments that optimize a measure of local similarity, the maximal segment pair (MSP) score. Recent mathematical results on the stochastic properties of MSP scores allow an analysis of the performance of this method as well as the statistical significance of alignments it generates. The basic algorithm is simple and robust; it can be implemented in a number of ways and applied in a variety of contexts including straightforward DNA and protein sequence database searches, motif searches, gene identification searches, and in the analysis of multiple regions of similarity in long DNA sequences. In addition to its flexibility and tractability to mathematical analysis, BLAST is an order of magnitude faster than existing sequence comparison tools of comparable sensitivity.
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            Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

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              The PRIDE database and related tools and resources in 2019: improving support for quantification data

              Abstract The PRoteomics IDEntifications (PRIDE) database (https://www.ebi.ac.uk/pride/) is the world’s largest data repository of mass spectrometry-based proteomics data, and is one of the founding members of the global ProteomeXchange (PX) consortium. In this manuscript, we summarize the developments in PRIDE resources and related tools since the previous update manuscript was published in Nucleic Acids Research in 2016. In the last 3 years, public data sharing through PRIDE (as part of PX) has definitely become the norm in the field. In parallel, data re-use of public proteomics data has increased enormously, with multiple applications. We first describe the new architecture of PRIDE Archive, the archival component of PRIDE. PRIDE Archive and the related data submission framework have been further developed to support the increase in submitted data volumes and additional data types. A new scalable and fault tolerant storage backend, Application Programming Interface and web interface have been implemented, as a part of an ongoing process. Additionally, we emphasize the improved support for quantitative proteomics data through the mzTab format. At last, we outline key statistics on the current data contents and volume of downloads, and how PRIDE data are starting to be disseminated to added-value resources including Ensembl, UniProt and Expression Atlas.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SoftwareRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: ResourcesRole: ValidationRole: Writing – review & editing
                Role: Formal analysisRole: SoftwareRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Funding acquisitionRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS Negl Trop Dis
                PLoS Negl Trop Dis
                plos
                plosntds
                PLoS Neglected Tropical Diseases
                Public Library of Science (San Francisco, CA USA )
                1935-2727
                1935-2735
                10 February 2021
                February 2021
                : 15
                : 2
                : e0009027
                Affiliations
                [1 ] Department of Veterinary Medicine, University of Bari, Bari, Italy
                [2 ] Department of Parasitology, ICB, Glycoproteomics Laboratory, University of São Paulo, Brazil
                [3 ] Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, Bari, Italy
                [4 ] IDEXX Laboratories, Inc., 1 IDEXX Drive, Westbrook, Maine, United States of America
                [5 ] Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Felestin Sq., Hamedan, Iran
                University of Utah, UNITED STATES
                Author notes

                We have read the journal's policy and the authors of this manuscript have the following competing interests: patent application filed and jointly submitted by the institution to which the following authors LMS, OD and PG are affiliated and/or from which the authors may benefit. RC is employee of IDEXX Laboratories, which funded partially the study. AG and PCL have no competing interests. IDEXX played a role in the review, approval and decision to publish of the manuscript.

                Author information
                https://orcid.org/0000-0001-9061-437X
                https://orcid.org/0000-0003-1816-548X
                https://orcid.org/0000-0003-0567-1076
                https://orcid.org/0000-0002-7518-476X
                Article
                PNTD-D-20-01390
                10.1371/journal.pntd.0009027
                7875354
                33566799
                b3b0e1cf-52d3-42af-9db8-147f4bbef9cf
                © 2021 Latrofa 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
                : 4 August 2020
                : 2 December 2020
                Page count
                Figures: 8, Tables: 4, Pages: 23
                Funding
                Funded by: IDEXX Laboratories
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo (BR)
                Award ID: FAPESP: 2014/06863-3, 2018/18257-1, 2018/15549-1
                Award Recipient :
                Funded by: “bolsa de produtividade” by CNPq
                Award ID: SNM (2017/04032-5)
                Award Recipient :
                Funded by: University of Bari
                Award Recipient :
                This study was funded by IDEXX Laboratories, Inc., Westbrook, Maine, USA.GP is supported by the São Paulo Research Foundation (FAPESP) (2014/06863-3, 2018/18257-1, 2018/15549-1), SNM (2017/04032-5), “bolsa de produtividade” by CNPq and Visiting Professor fellowship from the University of Bari (2017-2019). IDEXX played a role in the review, approval and decision to publish of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Dogs
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
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                Biology and Life Sciences
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                Onchocerca Volvulus
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                Onchocerca Volvulus
                Biology and Life Sciences
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                Biology and Life Sciences
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                Onchocerca Volvulus
                Biology and Life Sciences
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                Onchocerca
                Biology and Life Sciences
                Zoology
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                Biochemistry
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                All relevant data are within the manuscript and its Supporting Information files.

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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