NADPH oxidase activity is associated with cardiac osteopontin and pro-collagen type I expression in uremia.

International audienceCardiovascular disease is a frequent complication inducing mortality in chronic kidney disease (CKD) patients, which can be determined by both traditional risk factors and non-traditional risk factors such as malnutrition and oxidative stress. This study aimed to investigate the role of oxidative stress in uremia-induced cardiopathy in an experimental CKD model. CKD was induced in Sprague-Dawley rats by a 4-week diet supplemented in adenine, calcium and phosphorous and depleted in proteins. CKD was associated with a 3-fold increase in superoxide anion production from the NADPH oxidase in the left ventricle, but the maximal activity of mitochondrial respiratory chain complexes was not different. Although manganese mitochondrial SOD activity decreased, total SOD activity was not affected and catalase or GPx activities were increased, strengthening the major role of NADPH oxidase in superoxide anion output. Superoxide anion output was associated with enhanced expression of osteopontin (×7.7) and accumulation of pro-collagen type I (×3.7). To conclude, the increased activity of NADPH oxidase during CKD is associated with protein modifications which could activate a pathway leading to cardiac remodelling

Triglycerides and glycated hemoglobin for screening insulin resistance in obese patients

International audienceOBJECTIVE: Assessment of insulin resistance (IR) is essential in non-diabetic patients with obesity. Thus study aims to identify the best determinants of IR and to propose an original approach for routine assessment of IR in obesity. DESIGN AND PATIENTS: All adult with obesity defined by a body mass index >=30kg/m2, evaluated in the Nutrition Department between January 2010 and January 2015 were included in this cross-sectional study. Patients with diabetes were excluded. IR was diagnosed according to the HOMA-IR. Based on a logistic regression, we determined a composite score of IR. We then tested the variables with a principal component analysis and a hierarchical clustering analysis. RESULTS: A total of 498 patients with obesity were included. IR was associated with grade III obesity (OR=2.6[1.6-4.4], p\textless0.001), HbA1c>=5.7% (OR=2.6[1.7-4.0], p\textless0.001), hypertriglyceridemia \textgreater1.7mmol/l (OR=3.0[2.0-4.5], p\textless0.001) and age (OR=0.98[0.96-0.99], p=0.002). Exploratory visual analysis using factor map and clustering analysis revealed that lipid and carbohydrates metabolism abnormalities were correlated with insulin resistance but not with excessive fat accumulation and low-grade inflammation. CONCLUSIONS: Our results highlight the interest of simple blood tests such as HbA1c and triglyceride determination, which associated with BMI, may be widely available tools for screening IR in obese patients

Enzymatic creatinine assays allowestimation of glomerular filtration rate in stages 1 and 2 chronic kidney disease using CKD-EPI equation

The National Kidney Disease Education Program group demonstrated that MDRD equation is sensitive to creatinine measurement error, particularly at higher glomerular filtration rates. Thus, MDRD-based eGFR above 60 mL/min/1.73 m2 should not be reported numerically. However, little is known about the impact of analytical error on CKD-EPI-based estimates. This study aimed at assessing the impact of analytical characteristics (bias and imprecision) of 12 enzymatic and 4 compensated Jaffe previously characterized creatinine assays on MDRD and CKD-EPI eGFR. In a simulation study, the impact of analytical error was assessed on a hospital population of 24 084 patients. Ability using each assay to correctly classify patients according to chronic kidney disease (CKD) stages was evaluated. For eGFR between 60 and 90 mL/min/1.73 m2, both equations were sensitive to analytical error. Compensated Jaffe assays displayed high bias in this range and led to poorer sensitivity/specificity for classification according to CKD stages than enzymatic assays. As compared to MDRD equation, CKD-EPI equation decreases impact of analytical error in creatinine measurement above 90 mL/min/1.73 m2. Compensated Jaffe creatinine assays lead to important errors in eGFR and should be avoided. Accurate enzymatic assays allow estimation of eGFR until 90 mL/min/1.73 m2 with MDRD and 120 mL/min/1.73 m2 with CKD-EPI equation.Peer reviewe

Bone Biomarkers Help Grading Severity of Coronary Calcifications in Non Dialysis Chronic Kidney Disease Patients

BACKGROUND: Osteoprotegerin (OPG) and fibroblast growth factor-23 (FGF23) are recognized as strong risk factors of vascular calcifications in non dialysis chronic kidney disease (ND-CKD) patients. The aim of this study was to investigate the relationships between FGF23, OPG, and coronary artery calcifications (CAC) in this population and to attempt identification of the most powerful biomarker of CAC: FGF23? OPG? METHODOLOGY/PRINCIPAL FINDINGS: 195 ND-CKD patients (112 males/83 females, 70.8 [27.4-94.6] years) were enrolled in this cross-sectional study. All underwent chest multidetector computed tomography for CAC scoring. Vascular risk markers including FGF23 and OPG were measured. Logistic regression analyses were used to study the potential relationships between CAC and these markers. The fully adjusted-univariate analysis clearly showed high OPG (≥10.71 pmol/L) as the only variable significantly associated with moderate CAC ([100-400[) (OR = 2.73 [1.03;7.26]; p = 0.04). Such association failed to persist for CAC scoring higher than 400. Indeed, severe CAC was only associated with high phosphate fractional excretion (FEPO(4)) (≥38.71%) (OR = 5.47 [1.76;17.0]; p = 0.003) and high FGF23 (≥173.30 RU/mL) (OR = 5.40 [1.91;15.3]; p = 0.002). In addition, the risk to present severe CAC when FGF23 level was high was not significantly different when OPG was normal or high. Conversely, the risk to present moderate CAC when OPG level was high was not significantly different when FGF23 was normal or high. CONCLUSIONS: Our results strongly suggest that OPG is associated to moderate CAC while FGF23 rather represents a biomarker of severe CAC in ND-CKD patients

Biomarkers of cardiovascular risk in chronic kidney disease

Les maladies cardiovasculaires apparaissent précocement au cours de l'insuffisance rénale chronique (IRC) et représentent la première cause de mortalité. La 1ère étape pour apprécier la relation entre risque cardiovasculaire et progression de l'IRC consiste à améliorer l'estimation du débit de filtration glomérulaire (DFG). Nous avons donc évalué l'impact des conditions analytiques de mesure de la créatininémie et de la cystatinémie sur l'estimation du DFG. Les créatinines IDMS traçables (enzymatique et Jaffe compensé) améliorent l'estimation du DFG. Cependant, les méthodes enzymatiques non sensibles aux pseudochromogènes doivent être préférées. Concernant la cystatine C, nos résultats soulignent l'absence de standardisation du dosage. Chez des patients IRC non dialysés (stade I à V), nous avons identifié l'ostéoprotégérine (OPG) comme marqueur biologique de la présence de calcifications vasculaires. In vitro, nous avons démontré que le stress oxydant, majoré en présence de sérum urémique, jouait un rôle clé dans la transdifférenciation des cellules musculaires lisses vasculaires en ost oblastes. La mortalité en dialyse reste élevée et est largement dépendante des maladies cardiovasculaires. Il nous a donc paru nécessaire de rechercher les marqueurs pronostics et/ou d'en suivre l'évolution en transplantation. En dialyse, malgré une épuration significative par hémodiafiltration, les peptides natriurétiques sont des marqueurs du remodelage ventriculaire. La combinaison "NT-proBNP-CRP" est un puissant facteur pronostic de mortalité cardiovasculaire en hémodialyse. Après transplantation rénale, les calcifications vasculaires se stabilisent chez la majorité des patients et les taux d'OPG diminuent précocement. Les taux d'OPG sont significativement plus élevés chez les patients dont les calcifications progressent. Toutefois, seule l'intensité des calcifications avant transplantation permet de prédire la progressionCardiovascular disease occurs in the early stage of chronic kidney disease (CKD) and is the leading cause of death. The first step, to appreciate the link between cardiovascular risk and CKD progression, is to improve glomerular filtration rate (GFR) estimation. We have therefore evaluated the impact of analytical conditions for creatinine and cystatin C measurement on estimated GFR. New creatinine ID-MS traceable methods (enzymatic and compensated Jaffe) improved estimation of GFR by predictive equations. However, enzymatic methods that are much less susceptible to interfere with non-creatinine chromogens may provide more reliable estimations of GFR. Regarding cystatin C, our results highlighted the lack of standardization. In non dialyzed CKD patients (stage I to V), we identified osteoprotegerin (OPG) as a biomarker for the presence of vascular calcification. In vitro, we demonstrated that oxidative stress, increased in the presence of uremi c serum, played a key role in the transdifferentiation of vascular smooth muscle cells into osteoblast-like cells. In dialysis, mortality is high and largely dependant on cardiovascular disease. We have therefore investigated prognostic markers and/or followed their evolution after transplantation. In dialysis, despite their removal by hemodiafiltration, natriuretic peptides could be potential markers of left ventricular remodelling. In addition, the combination of high CRP and circulating NT-proBNP dramatically impaired the hemodialysis survival rate. After renal transplantation, stabilization of vascular calcification was observed in the majority of patients and OPG levels are dramatically reduced. Despite a higher baseline OPG level in progressors vs. non-progressors patients, post transplant vascular calcification progression was only predicted by baseline score

Biomarqueurs du risque cardiovasculaire en insuffisance rénale chronique

Les maladies cardiovasculaires apparaissent précocement au cours de l'insuffisance rénale chronique (IRC) et représentent la première cause de mortalité. La 1ère étape pour apprécier la relation entre risque cardiovasculaire et progression de l'IRC consiste à améliorer l'estimation du débit de filtration glomérulaire (DFG). Nous avons donc évalué l'impact des conditions analytiques de mesure de la créatininémie et de la cystatinémie sur l'estimation du DFG. Les créatinines IDMS traçables (enzymatique et Jaffe compensé) améliorent l'estimation du DFG. Cependant, les méthodes enzymatiques non sensibles aux pseudochromogènes doivent être préférées. Concernant la cystatine C, nos résultats soulignent l'absence de standardisation du dosage. Chez des patients IRC non dialysés (stade I à V), nous avons identifié l'ostéoprotégérine (OPG) comme marqueur biologique de la présence de calcifications vasculaires. In vitro, nous avons démontré que le stress oxydant, majoré en présence de sérum urémique, jouait un rôle clé dans la transdifférenciation des cellules musculaires lisses vasculaires en ost oblastes. La mortalité en dialyse reste élevée et est largement dépendante des maladies cardiovasculaires. Il nous a donc paru nécessaire de rechercher les marqueurs pronostics et/ou d'en suivre l'évolution en transplantation. En dialyse, malgré une épuration significative par hémodiafiltration, les peptides natriurétiques sont des marqueurs du remodelage ventriculaire. La combinaison "NT-proBNP-CRP" est un puissant facteur pronostic de mortalité cardiovasculaire en hémodialyse. Après transplantation rénale, les calcifications vasculaires se stabilisent chez la majorité des patients et les taux d'OPG diminuent précocement. Les taux d'OPG sont significativement plus élevés chez les patients dont les calcifications progressent. Toutefois, seule l'intensité des calcifications avant transplantation permet de prédire la progressionCardiovascular disease occurs in the early stage of chronic kidney disease (CKD) and is the leading cause of death. The first step, to appreciate the link between cardiovascular risk and CKD progression, is to improve glomerular filtration rate (GFR) estimation. We have therefore evaluated the impact of analytical conditions for creatinine and cystatin C measurement on estimated GFR. New creatinine ID-MS traceable methods (enzymatic and compensated Jaffe) improved estimation of GFR by predictive equations. However, enzymatic methods that are much less susceptible to interfere with non-creatinine chromogens may provide more reliable estimations of GFR. Regarding cystatin C, our results highlighted the lack of standardization. In non dialyzed CKD patients (stage I to V), we identified osteoprotegerin (OPG) as a biomarker for the presence of vascular calcification. In vitro, we demonstrated that oxidative stress, increased in the presence of uremi c serum, played a key role in the transdifferentiation of vascular smooth muscle cells into osteoblast-like cells. In dialysis, mortality is high and largely dependant on cardiovascular disease. We have therefore investigated prognostic markers and/or followed their evolution after transplantation. In dialysis, despite their removal by hemodiafiltration, natriuretic peptides could be potential markers of left ventricular remodelling. In addition, the combination of high CRP and circulating NT-proBNP dramatically impaired the hemodialysis survival rate. After renal transplantation, stabilization of vascular calcification was observed in the majority of patients and OPG levels are dramatically reduced. Despite a higher baseline OPG level in progressors vs. non-progressors patients, post transplant vascular calcification progression was only predicted by baseline score.MONTPELLIER-BU Pharmacie (341722105) / SudocSudocFranceF

Estimation of glomerular filtration rate using cystatin C.

International audienceBlood concentration of cystatin C is independent of muscle mass and tubular secretion. It can be used, in the absence of a reference method, as an alternative marker to creatinine for the evaluation of renal function and the estimation of glomerular filtration rate (GFR). Both particle-enhanced immunonephelemetry (PENIA) or immunoturbidimetry (PETIA) methods are available to determine cystatin C. From an analytical point of view, it is recommended to use methods whose calibration is traceable to the reference material (ERM-DA471/IFCC) and to report an estimated GFR based on cystatin C. The main equations used are those developed in 2012 by the group "Chronic kidney disease epidemiology collaboration (CKD-EPI)" for adults and those published by Schwartz in 2012 for children. National and international recommendations suggest using a cystatin C-based GFR estimate as a confirmatory test in the clinical settings where the relationship between creatinine production and muscular mass impairs the clinical performance of creatinine. The indications retained by the working group were graded according to the level of recommendations. The essential indications are the estimation and/or the monitoring of renal function in children and adolescents due to rapid changes in muscle mass; in patients with impaired muscle mass and in patients with an alteration of tubular secretion of creatinine (essentially iatrogenic effects)

Comparison of four immunoassays to an HPLC method for the therapeutic drug monitoring of methotrexate: Influence of the hydroxylated metabolite levels and impact on clinical threshold.

International audienceObjectives: Methotrexate requires therapeutic drug monitoring in oncology because of narrow therapeutic index, especially the metabolite 7-hydroxymethotrexate exhibits nephrotoxicity. The goal of this study was to evaluate different assays and their impact on clinical decisions.Methods: Following routine measurement with Abbott TDxFLx® assay (MTX-TDX), 62 samples were analysed on Architect®i1000 (MTX-ARCHI), Xpand® (ARK/XPND), Indiko® (ARK/INDI), and HPLC (MTX-HPLC) as the reference method. The influence of 7-hydroxymethotrexate was explored on ARK reagent to document the cause of the observed bias. ROC curves were built to study the impact of the method on the discharge thresholds for the patients at three levels.Results: Total imprecision was below 2.60% for the methotrexate-ARCHI and close to 10% for both ARK assays for plasma pools. The correlation coefficients were 0.93, 0.93, 0.89 and 0.95, the Bland–Altman difference plot revealed a bias of 0.075, 0.037, 0.049 and –0.002, and the number of results exceeding the TE criteria of 0.1 µM was 17 (27%), 13 (21%), 15 (24%) and 15 (24%) for MTX-TDX, ARK/INDI, ARK/XPND and MTX-ARCHI, respectively. Cross reactivity with 7-hydroxymethotrexate was between 1 and 9%. Overestimation of methotrexate concentration was between –4% and +32%. The most robust clinical level was found to be the highest level (0.2 µM) with ROC curves.Conclusions: The authors found the best results for imprecision with chemiluminescent microparticle immunoassay method on methotrexate-ARCHI, with bias below to the RICOS recommendations and best correlation to the reference method. Impact on the threshold values for clinical decision need to be clearly exposed to clinicians

Determination of hemolysis cut-offs for biochemical and immunochemical analytes according to their value.

International audienceBackground All general biochemistry instruments allow the measure of hemolysis index (HI), and suppliers provide an acceptable HI for each assay without consideration of the analyte value or its clinical application. Our first objective was to measure the impact of hemolysis degree on plasma biochemical and immunochemical analytes to determine the maximum allowable HI for each of them using four calculation methods as significant bias in comparison to manufacturer's data. The second objective was to assess whether the maximum allowable HI varied according to the analyte values. Methods Twenty analytes were measured in hemolyzate-treated plasma to determine the HI leading to a significant change compared to baseline value. Analytes were assessed at one (3 analytes), two (5 analytes) and three (12 analytes) values according to their sensitivity to hemolysis and their clinical impact. We used four calculation methods as significant limit from baseline value: the total change limit (TCL), the 10% change (10%Δ), the analytical change limit and the reference change value. Results Allowable HI was significantly different according to the threshold chosen for most analytes and was also dependent on the analyte value for alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatine kinase, iron, haptoglobin and high sensitivity troponin T. No hemolysis interference was observed for albumin, creatinine, C-reactive protein, and procalcitonin even at an HI value of 11 g/L. Conclusions This study highlights that TCL is the most appropriate calculation method to determine allowable HI in practice for biochemical and immunochemical parameters using Cobas 8000© from Roche Diagnostics. In addition, different allowable HI were found according to analyte value leading to optimization of resampling to save time in patient care

Analytical evaluation of the performances of a new procalcitonin immunoassay

International audienceNo abstract availabl