Prognosis of Peritoneal Dialysis Compared to Hemodialysis in Patient with End-Stage Renal Disease

End-stage Renal Disease (ESRD) is the terminal stage of Chronic Kidney Disease, where the function of the failing kidney must be substituted with Renal Replacement Therapy (RRT). There are two forms of RRT; Peritoneal Dialysis (PD) and Hemodialysis (HD. However, the issue of which method provide a better survival for patient remains an interesting topic to date. This paper aims to provide evidence on whether PD provides better survival compared to HD in a patient with ESRD. Systematic search was done using two databases; Pubmed® and Scopus®. Cohort studies were selected as appropriate study design to answer a prognosis question. Two restrospective cohorts and one prospective cohort study are relevant for this report. Two studies demonstrated survival advantage of PD over HD described by Relative Risk of Mortality of 0.398 and 0.49. The last study showed worse survival of PD patients compared to HD (RR=1.82). The difference in survival in the last study may be attributed to the fact that patients undergoing PD has worse baselinecharacteristics. PDand HD bring about comparable survival in ESRD patients

The Association Between Biliary Atresia and Cytomegalovirus Infection

Perinatal infection of cytomegalovirus (CMV) may cause cholestasis resembling biliary atresia. CMV infection is found in patients with biliary atresia. This simultaneous occurrence of biliary atresia and CMV infection prompted speculation that CMV may contribute to the progression of biliary atresia. This report has the objective to obtain the evidence regarding the association between biliary atresia and CMV infection. Two databases were searched to obtain the evidence: PubMed and Scopus. The study design which was selected for this report was case control due to its relevance to answer the clinical question. There were two case control studies which are appropriate to answer the clinical question. Both studies showed that biliary atresia is associated with CMV infection (OR>1). Biliary atresia is associated with CMV infection at the time of diagnosis, therefore the presence of CMV infection in neonatal cholestasis should not delay the investigation towards biliary atresia

Age-related mitochondrial DNA depletion and the impact on pancreatic beta cell function

Type 2 diabetes is characterised by an age-related decline in insulin secretion. We previously identified a 50% age-related decline in mitochondrial DNA (mtDNA) copy number in isolated human islets. The purpose of this study was to mimic this degree of mtDNA depletion in MIN6 cells to determine whether there is a direct impact on insulin secretion. Transcriptional silencing of mitochondrial transcription factor A, TFAM, decreased mtDNA levels by 40% in MIN6 cells. This level of mtDNA depletion significantly decreased mtDNA gene transcription and translation, resulting in reduced mitochondrial respiratory capacity and ATP production. Glucose-stimulated insulin secretion was impaired following partial mtDNA depletion, but was normalised following treatment with glibenclamide. This confirms that the deficit in the insulin secretory pathway precedes K+ channel closure, indicating that the impact of mtDNA depletion is at the level of mitochondrial respiration. In conclusion, partial mtDNA depletion to a degree comparable to that seen in aged human islets impaired mitochondrial function and directly decreased insulin secretion. Using our model of partial mtDNA depletion following targeted gene silencing of TFAM, we have managed to mimic the degree of mtDNA depletion observed in aged human islets, and have shown how this correlates with impaired insulin secretion. We therefore predict that the age-related mtDNA depletion in human islets is not simply a biomarker of the aging process, but will contribute to the age-related risk of type 2 diabetes

TFAM mRNA silencing induces mtDNA depletion 72 h post transfection.

<p>MIN6 cells were transfected with TFAM-193, TFAM-429 or Scrambled siRNA probes, or with no siRNA (Shocked). TFAM mRNA expression was quantified relative to reference gene <i>B2M</i> by real-time PCR at 48 h (A) and 72 h (B) post transfection. mtDNA depletion was also measured by real-time PCR, using mitochondrial encoded <i>ND5</i> relative to nuclear encoded <i>GAPDH</i> at 48 h (C) and 72 h (D) post transfection. All results normalised to Scrambled negative control. Experiment repeated once (C), twice (A) or 4 times (B, D) in triplicate. Data presented are means ± SEM (SD in (C)). * p&lt;0.05, *** p&lt;0.001.B2M, β2 Microglobulin; GAPDH, Glyceraldehyde-3-Phosphate Dehydrogenase; ND5, NADH Dehydrogenase 5; TFAM, Mitochondrial Transcription Factor A.</p

The effect of partial mtDNA depletion on mitochondrial function.

<p>Cells were harvested 72 h post transfection and oxygen consumption rate (OCR) was measured using the Seahorse XF24 Analyzer. OCR in TFAM-429 cells (n = 8) was severely impaired compared to that of Scrambled (n = 8) and Shocked (n = 8) control cells (A). Mitochondrial activity was measured following injection of oligomycin, an inhibitor of Complex V (ATP Synthase), followed by two sequential injections of FCCP to uncouple respiration and induce maximal respiration, and finally antimycin, an inhibitor of Complex III (Ubiquinol-Cytochrome <i>c</i> Reductase) preventing electron transfer and subsequently abolishing the proton gradient required for ATP synthesis. Basal and maximal respiratory capacity (B) and ATP synthesis by oxidative phosphorylation (OXPHOS) (C) were calculated as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0115433#pone.0115433-Bonnen1" target="_blank">[30]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0115433#pone.0115433-Brand2" target="_blank">[32]</a>. Data were normalised to protein concentration and are presented means ± SEM. * p&lt;0.5, ** p&lt;0.001, *** p&lt;0.0001.</p

The effect of glibenclamide on insulin secretion following <i>TFAM</i> silencing-induced mtDNA depletion.

<p>Seventy two hours post transfection cells were stimulated with 3 mM or 25 mM glucose, supplemented with or without 0.1 µM glibenclamide. Insulin secretion was determined by insulin ELISA and normalised to whole cell protein content. Data shown are from 4 separated experiments performed in triplicate, and are normalised to Scrambled negative control cells stimulated with 3 mM glucose without glibenclamide. Data presented are means ± SEM. * p&lt;0.05, ** p&lt;0.01.</p

The effect of partial mtDNA depletion on mitochondrial gene transcription and protein translation.

<p>Mitochondrial DNA was depleted following <i>TFAM</i> silencing and COX1 mRNA expression was quantified relative to reference gene <i>B2M</i> 72 h post transfection (A). Protein was extracted 72 h post transfection and analysed by western blotting, probing for COX1, SDH70, and β-Actin proteins. Protein bands were quantified by densitometry, and optical density readings used to calculate the ratio of COX1 (B) and SDH70 (C) mitochondrial proteins relative to β-Actin loading control. A representative blot is shown in (D). Data in (A) are normalised to Scrambled control cells. Both experiments repeated 3 times, with each experimental repeat performed in triplicate (A) or duplicate (B, C). Data presented are means ± SEM. * p&lt;0.05, ** p&lt;0.01, *** p&lt;0.001. COX1, Cytochrome <i>c</i> Oxidase 1; SDH70, Succinate Dehydrogenase 70 kDa subunit.</p

The effect of partial mtDNA depletion on glucose-stimulated insulin secretion.

<p>Seventy two hours post transfection cells were stimulated with basal (3 mM) or high (25 mM) glucose concentrations. Insulin secretion (A) and insulin content (B) were determined by insulin ELISA and normalised to protein content. Data normalised to 3 mM glucose stimulated Scrambled control cells. mtDNA levels (C) and <i>Ins1</i> insulin gene expression (D) were quantified and normalised to the Scrambled control. Data shown are from 9 (A) or 3 (B, C, D) separate experiments, each performed in triplicate. Data presented are means ± SEM. ** p≤0.01, *** p&lt;0.001.</p