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      Serum amyloid A is a positive acute phase protein in Russian sturgeon challenged with Aeromonas hydrophila

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          Abstract

          The immune system of sturgeons, one of the most ancient and economically valuable fish worldwide, is poorly understood. The lack of molecular tools and data about infection biomarkers hinders the possibility to monitor sturgeon health during farming and detect infection outbreaks. To tackle this issue, we mined publicly available transcriptomic datasets and identified putative positive acute-phase proteins (APPs) of Russian sturgeons that could be induced by a bacterial infection and monitored using non-invasive methods. Teleost literature compelled us to focus on five promising candidates: hepcidin, a warm acclimation associated hemopexin, intelectin, serum amyloid A protein (SAA) and serotransferrin. Among them, SAA was the most upregulated protein at the mRNA level in the liver of sturgeons challenged with heat-inactivated or live Aeromonas hydrophila. To assess whether this upregulation yielded increasing SAA levels in circulation, we developed an in-house ELISA to quantify SAA levels in sturgeon serum. Circulating SAA rose upon bacterial challenge and positively correlated with hepatic saa expression. This is the first time serum SAA has been quantified in an Actinopterygii fish. Since APPs vary across different fish species, our work sheds light on sturgeon acute-phase response, revealing that SAA is a positive APP with potential value as infection biomarker.

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding

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              Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

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                Author and article information

                Contributors
                avillarino@fcien.edu.uy
                aferreira@fcien.edu.uy
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                17 December 2020
                17 December 2020
                2020
                : 10
                : 22162
                Affiliations
                [1 ]GRID grid.11630.35, ISNI 0000000121657640, Unidad de Inmunología Asociada al Instituto de Química Biológica, Facultad de Ciencias - Área de Inmunología, Departamento de Biociencias, Facultad de Química, , Universidad de la República, ; CP 11600 Montevideo, Uruguay
                [2 ]GRID grid.11630.35, ISNI 0000000121657640, Instituto de Investigaciones Pesqueras, Facultad de Veterinaria, , Universidad de la República, ; CP 11300 Montevideo, Uruguay
                [3 ]GRID grid.11630.35, ISNI 0000000121657640, Sección Bioquímica y Biología Molecular, Facultad de Ciencias, , Universidad de la República, ; CP 11400 Montevideo, Uruguay
                Article
                79065
                10.1038/s41598-020-79065-9
                7746741
                32001736
                1a4376be-5477-4ec9-9d9a-8a3d065dac62
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 17 September 2020
                : 2 December 2020
                Funding
                Funded by: ANII
                Award ID: FPA _1_2013_1_13470
                Award Recipient :
                Funded by: Comisión Académica de Posgrado, CAP
                Funded by: Sistema Nacional de Investigadores, Uruguay
                Funded by: FundRef http://dx.doi.org/10.13039/501100006048, Universidad de la República Uruguay;
                Funded by: Agencia Nacional de Investigación e Innovación (ANII)
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                Uncategorized
                animal biotechnology,applied immunology,biotechnology,immunology,innate immunity
                Uncategorized
                animal biotechnology, applied immunology, biotechnology, immunology, innate immunity

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