Excessive accumulation of extracellular matrix (ECM) is the hallmark of fibrotic diseases. In the kidney, it is the final common pathway of prevalent diseases, leading to chronic renal failure. While cytokines such as TGF-β play a fundamental role in myofibroblast transformation, recent work has shown that mitochondrial dysfunction and defective fatty acid oxidation (FAO), which compromise the main source of energy for renal tubular epithelial cells, have been proposed to be fundamental contributors to the development and progression of kidney fibrosis. MicroRNAs (miRNAs), which regulate gene expression post-transcriptionally, have been reported to control renal fibrogenesis. To identify miRNAs involved in the metabolic derangement of renal fibrosis, we performed a miRNA array screen in the mouse model of unilateral ureteral obstruction (UUO). MiR-150-5p and miR-495-3p were selected for their link to human pathology, their role in mitochondrial metabolism and their targeting of the fatty acid shuttling enzyme CPT1A. We found a 2- and 4-fold upregulation of miR-150-5p and miR-495-5p, respectively, in both the UUO and the folic acid induced nephropathy (FAN) models, while TGF-β1 upregulated their expressions in the human renal tubular epithelial cell line HKC-8. These miRNAs synergized with TGF-β regarding its pro-fibrotic effect by enhancing the fibrosis-associated markers Acta2, Col1α1 and Fn1. Bioenergetics studies showed a reduction of FAO-associated oxygen consumption rate (OCR) in HKC-8 cells in the presence of both miRNAs. Consistently, expression levels of their mitochondrial-related target genes CPT1A, PGC1α and the mitochondrial transcription factor A (TFAM), were reduced by half in renal epithelial cells exposed to these miRNAs. By contrast, we did not detect changes in mitochondrial mass and transmembrane potential (ΔѰm) or mitochondrial superoxide radical anion production. Our data support that miR-150 and miR-495 may contribute to renal fibrogenesis by aggravating the metabolic failure critically involved in tubular epithelial cells, ultimately leading to fibrosis.

Patients with advanced chronic kidney disease (CKD) exhibit higher prevalence of coronary artery calcification (CaC) than general population. CaC has been proposed as a risk factor for mortality in end-stage CKD, but most studies in the field are based on short-term follow-up. We conducted a cohort, 10-year prospective longitudinal study of consecutive cases referred to the renal unit. A non-enhanced multislice coronary computed tomography was performed at baseline. CaC was assessed by Agatston method. Patients were stratified according to their CaC score: severe calcification group (CaCs< 400 HU) and mild-moderate calcification group (CaCs≥400 HU). The overall and cardiovascular (CV) mortality, CV events, and factors potentially associated with CaC development were recorded. 137 patients with advanced CKD were enrolled and provided consent. Overall mortality rate was 58%; 40% due to CV events. The rate of overall mortality in the severe calcification group was 75%, and 30% in the low calcification group, whereas the rate of CV mortality was 35% vs. 6%, respectively (p < 0.001). The severe calcification group was older, had higher prevalence of type 2 diabetes mellitus, former cardiologic events, and lower albumin serum levels than the mild-moderate calcification group. In a multivariate Cox model, severe CaC was a significant predictor of CV mortality (HR 5.01; 95%CI 1.28 to 19.6, p = 0.02). Among advanced CKD, there was a significantly increase of CV mortality in patients with severe CaCs during a 10-year follow-up period. CaCs could be a useful prognostic tool to predict CV mortality risk in CKD patients.

A patient suffering from advanced chronic renal disease undergoes several dialysis sessions on different dates. Several clinical parameters are monitored during the different hours of any of these sessions. These parameters, together with the information provided by other parameters of analytical nature, can be very useful to determine the probability that a patient may suffer from hypotension during the session, which should be specially watched since it represents a proven factor of possible mortality. However, the analytical information is not always available to the healthcare personnel, or it is far in time, so the clinical parameters monitored during the session become key to the prevention of hypotension. This article presents an investigation to predict the appearance of hypotension during a dialysis session, using predictive models trained from a large dialysis database, which contains the clinical information of 98,015 sessions corresponding to 758 patients. The prediction model takes into account up to 22 clinical parameters measured five times during the session, as well as the gender and age of the patient. This model was trained by means of machine learning classifiers, providing a success in the prediction higher than 80%.

Extracellular matrix (ECM) components, through specific peptide motifs such as Arg‐Gly‐Asp (RGD), interact with integrins and can modify the behavior of cells. Transforming growth factor‐β1 (TGF‐β1) is the main cytokine involved in the synthesis of ECM proteins. We analyzed the effect of a RGD‐containing peptide, as Arg‐Gly‐Asp‐Ser (RGDS), on the regulation of TGF‐β1 secretion in cultured human mesangial cells. We found that RGDS increased mRNA expression and secretion of TGF‐β1 by stimulating the TGF‐β1 gene promoter. This effect was dependent on the interaction of RGDS with integrins. We evaluated the signaling pathways implicated in TGF‐β1 production by analyzing the effect of RGDS on kinase‐related integrins. RGDS stimulated tyrosine phosphorylation as well as integrin‐linked kinase (ILK) activity. However, tyrosine kinase inhibitors did not prevent the RGDS effect. In contrast, the inhibition of ILK by cell transfection with a kinase dead‐ILK completely abolished the increased TGF‐β1 secretion and promoter activity in the presence of RGDS. Thus RGDS modulates the secretion of TGF‐β1, probably through increased synthesis by interacting with integrins and activating ILK. This supports a role for ECM components in the regulation of their own secretion.

This article proposes a new framework for a Cloud-based eHealth platform concept focused on Cloud computing environments, since current and emerging approaches using digital clinical history increasingly demonstrate their potential in maintaining the quality of the benefits in medical care services, especially in computer-assisted clinical diagnosis within the field of infectious diseases and due to the worsening of chronic pathologies. Our objective is to evaluate and contrast the performance of the architectural patterns most commonly used for developing eHealth applications (i.e., service-oriented architecture (SOA) and microservices architecture (MSA)), using as reference the quantitative values obtained from the various performance tests and their ability to adapt to the required software attribute (i.e., versatile high-performance). Therefore, it was necessary to modify our platform to fit two architectural variants. As a follow-up to this activity, corresponding tests were performed that showed that the MSA variant functions better in terms of performance and response time compared to the SOA variant; however, it consumed significantly more bandwidth than SOA, and scalability in SOA is generally not possible or requires significant effort to be achieved. We conclude that the implementation of SOA and MSA depends on the nature and needs of organizations (e.g., performance or interoperability).

Since mesangial and endothelial cells interact in the kidney, the present experiments were designed to analyze the ability of human mesangial cells (HMC) to modulate endothelin-1 (ET-1) synthesis by human umbilical vein endothelial cells (HuVEC). The supernatants of HuVEC/HMC contained significantly lower amounts of ET-1 than those of HuVEC alone. This effect was not due to a decreased prepro-ET-1 mRNA expression and was only partially the consequence of HMC-dependent ET-1 degradation. Therefore, we tested the influence of the coculture on endothelin-converting enzyme-1 (ECE-1), and found a significant reduction of its mRNA and protein levels as well as a decreased activity in HuVEC/HMC as compared to HuVEC alone. Using a pharmacological blockade approach (sulotrobam, BN52021, losartan or catalase), losartan was shown to completely abolish down-regulation of ECE-1 observed in HuVEC/HMC. Angiotensin II (AII) induced a dose and time-dependent inhibition of ECE-1 expression in HuVEC. These results support the importance of cross-talk among different cell types in the regulation of vascular or renal function. ET-1, and particularly ECE-1, might constitute a target in this regulation. In addition, locally synthesized AII could be one of the mediators involved in the down-regulation of ECE-1.