How to assess the role of Pt and Zn in the nephrotoxicity of Pt anti-cancer drugs?: An investigation combining μXRF and statistical analysis. Part II: Clinical application

International audienceIn this contribution, an approach developed previously for mice is used for human biopsy. In the case of patient 1, Pt detection is performed 6 days after the last oxaliplatin infusion, while for patient 2, the biopsy was performed more than 15 days after his first platin infusion and several dialysis. Even for these biological samples, experiments show that synchrotron mediated mXRF is a suitable tool to detect Pt in kidney biopsy, and thus probably for any organ exposed to Pt. Therefore, mXRF could also be of major interest to decipher the mechanism beyond Pt induced neurotoxicity, ototoxicity on human biopsy. Pharmacoavailability of chemotherapies is a major concern because some treatment failures are explained by poor tumor penetration of the active molecule. mXRF could be an elegant way to map the distribution of Pt inside cancerous cells at the micrometer scale. Pt and Zn are only two of the numerous trace elements that mXRF can detect; heavy metal intoxication diagnosis and the toxicity mechanism probably could also benefit from this innovative technique

Arrêt cardio-respiratoire en hémodialyse chronique : facteurs de risque, prévention et conduite à tenir en 2015

International audiencePatients undergoing hemodialysis have a 10 to 20 times higher risk of sudden cardiac arrest (SCA) than the general population. Sudden cardiac death is a rare event (approximately 1 event per 10,000 sessions) but has a very high mortality rate. Epidemiological data comes almost exclusively from North American studies; there is a great lack of European data on the subject. Ventricular arrhythmia is the main mechanism of sudden cardiac deaths in dialysis patients. These patients develop increased sensitivity mainly due to a high prevalence of severe ischemic heart disease and left ventricular hypertrophy and to a frequent trigger event: electrolytic and plasma volume shifts during dialysis sessions. Unfortunately, accurate predictive markers of SCA do not exist, however some primary prevention trials using beta-blockers or angiotensin II receptor blockers are encouraging, while the use of implantable cardioverter defibrillators in the population of chronic dialysis patients remains controversial. Identification of patients at risk, minimizing trigger events such as electrolytic shifts and improving team skills in the diagnosis and initial resuscitation with the latest recommendations from 2010 seem necessary to reduce incidence and improve survival in this high risk population. Organization of European studies would also allow a more accurate view of this reality in our dialysis units.Le patient en hémodialyse chronique a un risque d’arrêt cardio-respiratoire (ACR) 10 à 20 fois plus élevé que la population générale. Il s’agit d’un événement rare (environ 1 événement par 10 000 séances), mais le taux de mortalité reste très élevé. Les données épidémiologiques actuelles proviennent quasi exclusivement d’études nord-américaines et l’on manque de données issues de registres européens sur le sujet. Les troubles du rythme ventriculaires graves sont le mode de survenue majoritaire de l’ACR dans cette population de dialysés chroniques qui cumule, d’une part, un terrain de sensibilité accrue de par la forte prévalence de la cardiopathie ischémique et l’hypertrophie ventriculaire gauche principalement, et, d’autre part, un événement déclencheur fréquent : les variations électrolytiques et volémiques inhérentes aux séances de dialyse. Les marqueurs prédictifs de la survenue d’ACR sont encore imparfaitement définis. Des études de prévention primaire médicamenteuse utilisant les bêtabloquants ou les bloqueurs du système rénine-angiotensine-aldostérone sont encourageantes, tandis que l’utilisation des défibrillateurs automatisés implantables dans la population de dialysés chroniques reste controversée. L’identification des patients à risque, la minimisation des événements déclencheurs comme les variations électrolytiques, et l’amélioration de la formation des équipes au diagnostic et aux premières mesures de réanimation grâce aux dernières recommandations datant de 2010 semblent nécessaires afin de diminuer l’incidence et d’améliorer la survie dans cette population à fort risque. L’organisation d’études européennes permettrait par ailleurs d’avoir une vision plus précise de cette réalité dans nos centres

Evidence of dipstick superiority over urine microscopy analysis for detection of hematuria

International audienceBackgroundThere is an unresolved debate on the best screening method for hematuria as a symptom of glomerulonephritis or urological malignancies. The urinary dipstick is generally considered as an imperfect surrogate for urine microscopy analysis.ResultsWe designed a study to compare urine microscopy analysis, urinary dipstick and flow cytometry, using controlled dilutions of blood in urine samples from volunteers collected in two different physiologically-relevant conditions (basal state and hyperhydration). We found that although all techniques were 100 % effective in detecting hematuria at basal state, these results were variably reproduced when testing the same final amount of hematuria in urine collected after hyperhydration. Our data shows a variable sensitivity for the detection of hematuria by urine microscopy analysis or flow cytometry, but not by urinary dipstick.ConclusionsUrinary dipstick qualifies as a better screening test for hematuria than urine microscopy analysis or flow cytometry, as it is sensitive and performs better in unstandardized conditions. It is universally available and also faster and cheaper than cytometric techniques

Evaluation of the ability of bone marrow derived cells to engraft the kidney and promote renal tubular regeneration in mice following exposure to cisplatin

International audienceIt has been suggested that bone marrow derived stem cells have the ability to engraft the kidney and improve the outcome of severe acute kidney injury (AKI) in mice exposed to high doses of cisplatin, providing hope for cancer patients in whom irreversible renal damage occasionally occurs following the use of this highly effective anti-tumor drug. We tested the therapeutic potential of bone marrow derived cells injected during the acute phase (day 3 after cisplatin administration) of experimentally-induced AKI in C57Bl6/J mice, characterized by massive tubular necrosis, apoptosis, and a low proliferation capacity. We failed to show any benefit of bone marrow derived cells versus a regular homogenate of intact renal cells, or normal saline. Using cell tracers and flow cytometry, we demonstrated that bone marrow derived cells did indeed home to the bone marrow of the recipients but failed to settle in the kidney. Conversely, renal cells homed to injured kidneys. However, neither cell therapy protected the animals against cisplatin-induced death. We therefore question the short-term efficacy of bone marrow derived cells used to repair established injuries of the tubular epithelium

Ex vivo analysis of renal proximal tubular cells

Background: Experimental models are inevitably a compromise between accurately reproducing a pathological situation and schematically simplifying it, which is intended to provide both relevance and conclusiveness. In-vivo models are very relevant, but multiple cell-types undergoing various changes may hinder the observation of individual molecular events. Results: Here, we describe a method for analyzing and isolating specific cell types from the kidney and studying the phenotype they have acquired in vivo. Using flow cytometry, immunofluorescence, and RT-PCR, we show that our method is suitable for studying and isolating proximal tubular cells with an anti Prominin-1 antibody. Kidneys are subjected to mechanical dissociation followed by flow-cytometry analysis. Hundreds of thousands of proximal tubular cells are then isolated by magnetic separation followed by direct analysis or primary cell culture. Using our method, we detect phenotypic changes in the proximal tubular cells after renal ischemia reperfusion, and we isolate the proximal tubular cells, with a purity over 80%. Conclusions: This method is efficient, quick, simple, and cheap, and should be useful for studying cell-type specific parameters after in vivo experimental studies. It is also a simple method to obtain a specific primary cell culture from any animal strain

Rapid Occurrence of Chronic Kidney Disease in Patients Experiencing Reversible Acute Kidney Injury after Cardiac Surgery

International audienceBackground: There is recent evidence to show that patients suffering from acute kidney injury are at increased risk of devel-oping chronic kidney disease despite the fact that surviving tubular epithelial cells have the capacity to fully regenerate renal tubules and restore renal function within days or weeks. The aim of the study was to investigate the impact of acute kidney injury on de novo chronic kidney disease.Methods: The authors conducted a retrospective population-based cohort study of patients initially free from chronic kidney disease who were scheduled for elective cardiac surgery with cardiopulmonary bypass and who developed an episode of acute kidney injury from which they recovered. The study was conducted at two French university hospitals between 2005 and 2015. These individuals were matched with patients without acute kidney injury according to a propensity score for develop-ing acute kidney injury.Results: Among the 4,791 patients meeting the authors’ inclusion criteria, 1,375 (29%) developed acute kidney injury and 685 fully recovered. Propensity score matching was used to balance the distribution of covariates between acute kidney injury and non- acute kidney injury control patients. Matching was possible for 597 cases. During follow-up, 34 (5.7%) had reached a diagnosis of chronic kidney disease as opposed to 17 (2.8%) in the control population (hazard ratio, 2.3; bootstrapping 95% CI, 1.9 to 2.6).Conclusions: The authors’ data consolidate the recent paradigm shift, reporting acute kidney injury asa strong risk factor for the rapid development of chronic kidney disease

Specific calpain inhibition protects kidney against inflammaging

International audienceCalpains are ubiquitous pro-inflammatory proteases, whose activity is controlled by calpastatin, their specific inhibitor. Transgenic mice over-expressing rabbit calpastatin (CalpTG) are protected against vascular remodelling and angiotensin II-dependent inflammation. We hypothesized that specific calpain inhibition would protect against aging-related lesions in arteries and kidneys. We analysed tissues from 2-months and 2-years-old CalpTG and wild-type mice and performed high throughput RNA-Sequencing of kidney tissue in aged mice. In addition, we analysed inflammatory response in the kidney of aged CalpTG and wild-type mice, and in both in vivo (monosodium urate peritonitis) and in vitro models of inflammation. At two years, CalpTG mice had preserved kidney tissue, less vascular remodelling and less markers of senescence than wild-type mice. Nevertheless, CalpTG mice lifespan was not extended, due to the development of lethal spleen tumors. Inflammatory pathways were less expressed in aged CalpTG mice, especially cytokines related to NF-κB and NLRP3 inflammasome activation. CalpTG mice had reduced macrophage infiltration with aging and CalpTG mice produced less IL-1α and IL-1β in vivo in response to inflammasome activators. In vitro, macrophages from CalpTG mice produced less IL-1α in response to particulate activators of inflammasome. Calpains inhibition protects against inflammaging, limiting kidney and vascular lesions related to aging