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      Peritoneal dialysis fluid biocompatibility impact on human peritoneal membrane permeability

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          ABSTRACT

          Background

          We have compared the effects of conventional lactate-based peritoneal dialysis fluid (CPDF) with respect to bicarbonate/lactate-based fluid on peritoneal ultrafiltration (UF) and peritoneal permeability, and on variations on gene expression in cells isolated from effluents of patients’ peritoneal bags.

          Methods

          This was a non-randomized sequential prospective study including all incident peritoneal dialysis (PD) patients ( n = 40) recruited in our centre. Peritoneal equilibration tests (PETs) were performed using CPDF or BPDF both containing 2.27% glucose during a 48-h interval in four different sequences. Gene expression variation of selected genes was measured by reverse transcription polymerase chain reaction in mesothelial cells obtained from the total drained fluid during the PET.

          Results

          In the overall study, the use of BPDF was associated with significantly lower mass transfer area coefficient for urea and creatinine, longer accelerated peritoneal examination test times for urea and creatinine, lower total pore area available for exchange over diffusion distance and lower UF. There were no differences in the gene expression of aquaporins 1–3, endothelial and inducible nitric oxide synthase ( NOS3 and NOS2), or interleukin-6. The SNAIL and E-CADHERIN gene expression normalized ratio was evaluated in peritoneal effluents of cells obtained from CPDF and BPDF. We observed that the SNAIL/E-CADHERIN mRNA ratio decreased when the dialysis sequence started with BPDF and went on to CPDF, but not when the sequence was the opposite.

          Conclusion

          This study shows that those patients who started PD treatment with BPDF were characterized by a better biocompatibility profile. BPDF associates with lower peritoneal permeability to small molecules and lower UF.

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          Most cited references18

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            Peritoneal dialysis and epithelial-to-mesenchymal transition of mesothelial cells.

            During continuous ambulatory peritoneal dialysis, the peritoneum is exposed to bioincompatible dialysis fluids that cause denudation of mesothelial cells and, ultimately, tissue fibrosis and failure of ultrafiltration. However, the mechanism of this process has yet to be elucidated. Mesothelial cells isolated from effluents in dialysis fluid from patients undergoing continuous ambulatory peritoneal dialysis were phenotypically characterized by flow cytometry, confocal immunofluorescence, Western blotting, and reverse-transcriptase polymerase chain reaction. These cells were compared with mesothelial cells from omentum and treated with various stimuli in vitro to mimic the transdifferentiation observed during continuous ambulatory peritoneal dialysis. Results were confirmed in vivo by immunohistochemical analysis performed on peritoneal-biopsy specimens. Soon after dialysis is initiated, peritoneal mesothelial cells undergo a transition from an epithelial phenotype to a mesenchymal phenotype with a progressive loss of epithelial morphology and a decrease in the expression of cytokeratins and E-cadherin through an induction of the transcriptional repressor snail. Mesothelial cells also acquire a migratory phenotype with the up-regulation of expression of alpha2 integrin. In vitro analyses point to wound repair and profibrotic and inflammatory cytokines as factors that initiate mesothelial transdifferentiation. Immunohistochemical studies of peritoneal-biopsy specimens from patients undergoing continuous ambulatory peritoneal dialysis demonstrate the expression of the mesothelial markers intercellular adhesion molecule 1 and cytokeratins in fibroblast-like cells entrapped in the stroma, suggesting that these cells stemmed from local conversion of mesothelial cells. Our results suggest that mesothelial cells have an active role in the structural and functional alteration of the peritoneum during peritoneal dialysis. The findings suggest potential targets for the design of new dialysis solutions and markers for the monitoring of patients. Copyright 2003 Massachusetts Medical Society
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              Hepatocyte growth factor induces cell scattering through MAPK/Egr-1-mediated upregulation of Snail.

              Hepatocyte growth factor/scatter factor (HGF) exerts several functions in physiological and pathological processes, among them the induction of epithelial cell scattering and motility. Its pivotal role in angiogenesis, tumor progression, and metastasis is evident; however, the underlying molecular mechanisms are still poorly understood. Here, we demonstrate that HGF induces scattering of epithelial cells by upregulating the expression of Snail, a transcriptional repressor involved in epithelial-mesenchymal transition (EMT). Snail is required for HGF-induced cell scattering, since shRNA-mediated ablation of Snail expression prevents this process. HGF-induced upregulation of Snail transcription involves activation of the mitogen-activated protein kinase (MAPK) pathway and requires the activity of early growth response factor-1 (Egr-1). Upon induction by Egr-1, Snail represses the expression of E-cadherin and claudin-3 genes. It also binds to the Egr-1 promoter and represses Egr-1 transcription, thereby establishing a negative regulatory feedback loop. These findings indicate that Snail upregulation by HGF is mediated via the MAPK/Egr-1 signaling pathway and that both Snail and Egr-1 play a critical role in HGF-induced cell scattering, migration, and invasion.
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                Author and article information

                Journal
                Clin Kidney J
                Clin Kidney J
                ckj
                Clinical Kidney Journal
                Oxford University Press
                2048-8505
                2048-8513
                December 2018
                25 June 2018
                25 June 2018
                : 11
                : 6
                : 881-888
                Affiliations
                [1 ]Unidad de Investigación, Hospital Universitario de Gran Canaria Dr Negrín, Las Palmas de Gran Canaria, Gran Canaria, Spain
                [2 ]Universidad de Las Palmas de Gran Canaria (ULPGC), Las Palmas de Gran Canaria, Gran Canaria, Spain
                [3 ]Servicio de Nefrología, Hospital Universitario de Gran Canaria Dr Negrín, Las Palmas de Gran Canaria, Gran Canaria, Spain
                Author notes
                Correspondence and offprint requests to: Nicanor Vega-Díaz; E-mail: nvegdia@ 123456gobiernodecanarias.org
                Article
                sfy043
                10.1093/ckj/sfy043
                6275447
                47f163ee-147e-46e4-8e03-6ee18128b87e
                © The Author(s) 2018. Published by Oxford University Press on behalf of ERA-EDTA.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                : 11 September 2017
                : 02 May 2018
                Page count
                Pages: 8
                Categories
                Peritoneal Dialysis

                Nephrology
                bicarbonate/lactate buffered solution,biocompatibility,epithelial-to-mesenchymal transition,mesothelial cells,peritoneal dialysis,peritoneal transport rate

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