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      Selenoprotein Transcript Level and Enzyme Activity as Biomarkers for Selenium Status and Selenium Requirements of Chickens ( Gallus gallus)

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          Abstract

          The NRC selenium (Se) requirement for broiler chicks is 0.15 μg Se/g diet, based primarily on weight gain and feed intake studies reported in 1986. To determine Se requirements in today’s rapidly growing broiler chick, day-old male chicks were fed Se-deficient basal diets supplemented with graded levels of Se (0, 0.025, 0.05, 0.075, 0.1, 0.2, 0.3, 0.5, 0.75, and 1.0 μg Se/g) as Na 2SeO 3 (5/treatment). Diets contained 15X the vitamin E requirement, and there were no gross signs of Se-deficiency. At 29 d, Se-deficient chicks weighed 62% of Se-supplemented chicks; 0.025 μg Se/g reversed this effect, indicating a minimum Se requirement of 0.025 μg Se/g diet for growth for male broiler chicks. Enzyme activities in Se-deficient chicks for plasma GPX3, liver and gizzard GPX1, and liver and gizzard GPX4 decreased dramatically to 3, 2, 5, 10 and 5%, respectively, of Se-adequate levels, with minimum Se requirements of 0.10–0.13 μg Se/g, and with defined plateaus above these levels. Pancreas GPX1 and GPX4 activities, however, lacked defined plateaus, with breakpoints at 0.3 μg Se/g. qPCR measurement of all 24 chicken selenoprotein transcripts, plus SEPHS1, found that SEPP1 in liver, GPX3 in gizzard, and SEPP1, GPX3 and SELK in pancreas were expressed at levels comparable to housekeeping transcripts. Only 33%, 25% and 50% of selenoprotein transcripts were down-regulated significantly by Se deficiency in liver, gizzard and pancreas, respectively. No transcripts could be used as biomarkers for supernutritional Se status. For export selenoproteins SEPP1 and GPX3, tissue distribution, high expression and Se-regulation clearly indicate unique Se metabolism, which may underlie tissues targeted by Se deficiency. Based on enzyme activities in liver, gizzard, and plasma, the minimum Se requirement in today’s broiler chick is 0.15 μg Se/g diet; pancreas data indicate that the Se requirement should be raised to 0.2 μg Se/g diet to provide a margin of safety.

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          Selenium: biochemical role as a component of glutathione peroxidase.

          When hemolyzates from erythrocytes of selenium-deficient rats were incubated in vitro in the presence of ascorbate or H(2)O(2), added glutathione failed to protect the hemoglobin from oxidative damage. This occurred because the erythrocytes were practically devoid of glutathione-peroxidase activity. Extensively purified preparations of glutathione peroxidase contained a large part of the (75)Se of erythrocytes labeled in vivo. Many of the nutritional effects of selenium can be explained by its role in glutathione peroxidase.
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            Experimental validation of novel and conventional approaches to quantitative real-time PCR data analysis.

            Real-time PCR is being used increasingly as the method of choice for mRNA quantification, allowing rapid analysis of gene expression from low quantities of starting template. Despite a wide range of approaches, the same principles underlie all data analysis, with standard approaches broadly classified as either absolute or relative. In this study we use a variety of absolute and relative approaches of data analysis to investigate nocturnal c-fos expression in wild-type and retinally degenerate mice. In addition, we apply a simple algorithm to calculate the amplification efficiency of every sample from its amplification profile. We confirm that nocturnal c-fos expression in the rodent eye originates from the photoreceptor layer, with around a 5-fold reduction in nocturnal c-fos expression in mice lacking rods and cones. Furthermore, we illustrate that differences in the results obtained from absolute and relative approaches are underpinned by differences in the calculated PCR efficiency. By calculating the amplification efficiency from the samples under analysis, comparable results may be obtained without the need for standard curves. We have automated this method to provide a means of streamlining the real-time PCR process, enabling analysis of experimental samples based upon their own reaction kinetics rather than those of artificial standards.
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              Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat.

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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                5 April 2016
                2016
                : 11
                : 4
                : e0152392
                Affiliations
                [1 ]College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People’s Republic of China
                [2 ]Department of Nutritional Sciences, University of Wisconsin, Madison, Wisconsin, United States of America
                Oklahoma State University, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: RAS. Performed the experiments: JLL RAS. Analyzed the data: JLL RAS. Wrote the paper: JLL RAS.

                Article
                PONE-D-15-49376
                10.1371/journal.pone.0152392
                4821606
                27045754
                29653995-4107-4651-b568-f2b62d6ca60f
                © 2016 Li, Sunde

                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
                : 18 November 2015
                : 14 March 2016
                Page count
                Figures: 9, Tables: 3, Pages: 24
                Funding
                This Research was supported by the National Institute of Food and Agriculture, United States Department of Agriculture ( www.csrees.usda.gov), Hatch projects 233618 and 1004389, by Northeast Agricultural University ( www.neau.edu.cn) Program for New Century Excellent Talents in University (No. NECT-1207-02) and New Century Excellent Talents in Heilongjiang Provincial University (No. 1252-NCET-009), and by the Wisconsin Alumni Foundation ( http://www.uwalumni.com) Selenium Nutrition Research Fund (No. 12046295). 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
                Nutrition
                Diet
                Medicine and Health Sciences
                Nutrition
                Diet
                Biology and Life Sciences
                Anatomy
                Endocrine System
                Pancreas
                Medicine and Health Sciences
                Anatomy
                Endocrine System
                Pancreas
                Biology and Life Sciences
                Anatomy
                Exocrine Glands
                Pancreas
                Medicine and Health Sciences
                Anatomy
                Exocrine Glands
                Pancreas
                Biology and life sciences
                Biochemistry
                Nucleic acids
                RNA
                Messenger RNA
                Biology and Life Sciences
                Biochemistry
                Biomarkers
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                Organisms
                Animals
                Vertebrates
                Amniotes
                Birds
                Fowl
                Gamefowl
                Chickens
                Biology and Life Sciences
                Organisms
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                Amniotes
                Birds
                Poultry
                Chickens
                Biology and Life Sciences
                Agriculture
                Livestock
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                Biology and Life Sciences
                Nutrition
                Nutritional Deficiencies
                Medicine and Health Sciences
                Nutrition
                Nutritional Deficiencies
                Physical sciences
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                Organic compounds
                Vitamins
                Vitamin E
                Physical sciences
                Chemistry
                Organic chemistry
                Organic compounds
                Vitamins
                Vitamin E
                Biology and Life Sciences
                Genetics
                Gene Expression
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