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      Study of the role of tumor necrosis factor-α (–308 G/A) and interleukin-10 (–1082 G/A) polymorphisms as potential risk factors to acute kidney injury in patients with severe sepsis using high-resolution melting curve analysis

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          Abstract

          Rational: Septic acute kidney injury (AKI) is a prevalent complication in intensive care units with an increased incidence of complications.

          Objective: The aim of the present study was to assess the use of high-resolution melting curve (HRM) analysis in investigating whether the genetic polymorphisms; –308 G/A of tumor necrosis factor-α (TNF-α), and –1082 G /A of Interleukin-10 (IL-10) genes may predispose patients diagnosed with severe sepsis to the development of AKI.

          Methods: One hundred and fifty patients with severe sepsis participated in the present study; only sixty-six developed AKI. Both polymorphisms were studied using HRM analysis.

          Main findings: The low producer genotype of both studied polymorphism of TNF-α and IL-10 genes was associated with AKI. Using logistic regression analysis, the low producer genotypes remained an independent risk factor for AKI. A statistically significant difference was detected between both studied groups as regards the low producer genotype in both TNF-α (–308 G/A) and interleukin-10 (IL-10) (–1082 G/A) polymorphisms being prevalent in patients developing AKI.

          Principle conclusions: The low producer genotypes of both TNF-α (–308 G/A) and IL-10 (–1082 G/A) polymorphisms could be considered a risk factor for the development of AKI in critically ill patients with severe sepsis, thus management technique implemented for this category should be modulated rescuing this sector of patients from the grave deterioration to acute kidney injury. Using HRM for genotyping proved to be a highly efficient, simple, cost-effective genotyping technique that is most appropriate for the routine study of large-scale samples.

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          Most cited references22

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          Recent advances in the pathophysiology of ischemic acute renal failure.

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            High-resolution DNA melting analysis for simple and efficient molecular diagnostics.

            High-resolution melting of DNA is a simple solution for genotyping, mutation scanning and sequence matching. The melting profile of a PCR product depends on its GC content, length, sequence and heterozygosity and is best monitored with saturating dyes that fluoresce in the presence of double-stranded DNA. Genotyping of most variants is possible by the melting temperature of the PCR products, while all variants can be genotyped with unlabeled probes. Mutation scanning and sequence matching depend on sequence differences that result in heteroduplexes that change the shape of the melting curve. High-resolution DNA melting has several advantages over other genotyping and scanning methods, including an inexpensive closed tube format that is homogenous, accurate and rapid. Owing to its simplicity and speed, the method is a good fit for personalized medicine as a rapid, inexpensive method to predict therapeutic response.
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              Polymorphic haplotypes of the interleukin-10 5' flanking region determine variable interleukin-10 transcription and are associated with particular phenotypes of juvenile rheumatoid arthritis.

              To determine the distribution of the interleukin-10 (IL-10) 5' flanking region haplotypes in children with arthritis and in controls, and to investigate the functional significance of each haplotype. Sequence-specific oligonucleotide probing was used to determine haplotype frequency. Transient transfection studies were used to investigate the transcription of reporter genes driven by each haplotype. Whole blood cultures were performed to assess IL-10 production by each genotype. Patients with arthritis involving >4 joints were more likely to have a genotype with an ATA haplotype than those whose arthritis remained restricted to <4 joints. This ATA haplotype was associated with lower transcriptional activity than the GCC haplotype (P = 0.02), and the ATA/ATA genotype was associated with lower IL-10 production under lipopolysaccharide stimulation than other genotypes (P < 0.02). The results of this study demonstrate the functional significance of the ATA haplotype and reveal a significant association of genotypes containing this haplotype with extended oligoarthritis.
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                Author and article information

                Journal
                Ren Fail
                Ren Fail
                IRNF
                irnf20
                Renal Failure
                Taylor & Francis
                0886-022X
                1525-6049
                2017
                27 October 2016
                : 39
                : 1
                : 77-82
                Affiliations
                [a ]Clinical Pathology Department, Faculty of Medicine, Alexandria University, Alexandria , Egypt;
                [b ]Critical Care Medicine Department, Faculty of Medicine, Alexandria University, Alexandria , Egypt
                Author notes
                CONTACT Doaa I. Hashad doaa_hashad2003@ 123456yahoo.com Associate Professor of Clinical Pathology, Department of Clinical Pathology, Faculty of Medicine, Alexandria University , Azarita, Alexandria, Egypt
                Article
                1244081
                10.1080/0886022X.2016.1244081
                6014343
                27788614
                fc319fdf-ee82-4977-983a-bc6c7678c286
                © 2016 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/Licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 09 June 2016
                : 25 August 2016
                : 16 September 2016
                Page count
                Pages: 6, Words: 4413
                Categories
                Laboratory Study

                Nephrology
                tnf,il-10,acute kidney injury,sepsis,hrm
                Nephrology
                tnf, il-10, acute kidney injury, sepsis, hrm

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