miRNAs in plasma and urine samples across various stages of diabeticnephropathy 16–20 .
Widely used techniques for identification of differentially expressed miRNAs, such as miRNA microarrays, suffer from a low accuracy due to low specificity of probe hybridization and variable sensitivity to detect less abundant miRNAs. The next-generation sequencing has a high-resolution potential, even though limited data is available in the relevant literature on its use as a discovery- based approach to identify enriched or dysregulated miRNAs in DN. The aim of the present study was to perform a comprehensive sequencing of plasma miRNAs in subjects with T2DM to identify
PS 14 Genetics and epidemiology of diabeticnephropathy
ACE I/D polymorphism predicts end stage renal disease and or mortality in type 1 diabetic patients except for those with already advanced nephropathy: the follow up of the Genesis/Genediab Studies
Cardiac autonomic neuropathy Antidiabetic drugs
A B S T R A C T
In diabetic patients undergoing surgery, we recommend assessing glycaemic control preoperatively by assessing glycated haemoglobin (HbA1c) levels and recent capillary blood sugar (glucose) levels, and to adjust any treatments accordingly before surgery, paying particular attention to speciﬁc complications of diabetes. Gastroparesis creates a risk of stasis and aspiration of gastric content at induction of anaesthesia requiring the use of a rapid sequence induction technique. Cardiac involvement can be divided into several types. Coronary disease is characterised by silent myocardial ischaemia, present in 30–50% of T2D patients. Diabetic cardiomyopathy is a real cause of heart failure. Finally, cardiac autonomic neuropathy (CAN), although rarely symptomatic, should be investigated because it causes an increased risk of cardiovascular events and a risk of sudden death. Several signs are suggestive of CAN, and conﬁrmation calls for close perioperative surveillance. Chronic diabetic kidney disease (diabeticnephropathy) aggravates the risk of perioperative acute renal failure, and we recommend measurement of the glomerular ﬁltration rate preoperatively. The ﬁnal step of the consultation concerns the management of antidiabetic therapy. Preoperative glucose infusion is not necessary if the patient is not receiving insulin. Non-insulin drugs are not administered on the morning of the intervention except for metformin, which is not administered from the evening before. The insulins are injected at the usual dose the evening before. The insulin pump is maintained until the patient arrives in the surgical unit. It should be remembered that insulin deﬁciency in a T1D patient leads to ketoacidosis within a few hours.
Personal history of dyslipidemia was slightly more frequent among relatives of index case subjects than others. This re- port confirms the data from a single- center family-based study showing a clustering of dyslipidemia with nephrop- athy (28). Familial clustering of lipid dis- turbances may partly account for the reported familial clustering of cardiovas- cular disease in type 1 diabetic patients with nephropathy (6,7,28,45). Lipid dis- turbances are well known to be associated with diabeticnephropathy (46,47), and prospective studies of type 1 diabetic pa- tients have shown that an adverse lipid profile is predictive of the worsening of renal disease (18,48) or the least regres- sion from microalbuminuria to nor- moalbuminuria (49). The adverse role of plasma lipids was recently found regard- ing renal and retinal complications in a longitudinal study (50). Several candidate genes involved in lipid metabolism could
5.1. Noninvasive Diagnosis of Primary MN
While the diagnosis of “primary” MN required a kidney biopsy until recently, the availability of assays for PLA2R antibodies has induced a dramatic change of the diagnostic strategy, owing to the very high specificity (99%) of anti-PLA2R antibody for the diagnosis of MN [ 59 , 60 ]. The 95% confidence interval for specificity is 0.96 to 1.0, which is comparable to the diagnostic performance of kidney biopsy. The added value of kidney biopsy was studied by the Mayo Clinic investigators in 97 patients who tested positive for PLA2R antibodies and had no evidence of a secondary cause of MN. Sixty of those 97 patients had an estimated glomerular filtration rate (eGFR) >60 mL/min/1.73 m 2 . In these patients, the kidney biopsy did not provide significant information that altered management; one patient had a superimposed diabeticnephropathy, and a second patient had a superimposed focal segmental glomerulosclerosis lesion. Among the 37 patients with primary MN and eGFR <60 mL/min/1.73 m 2 additional findings included acute interstitial nephritis (n=1), acute tubular necrosis (n = 1), FSGS (n = 2) with cellular crescents in one case, and diabeticnephropathy (n = 1). These findings were corroborated by Wiech et al. [ 61 ] who showed that only twelve (6%) of the 194 PLA2R1-Ab positive patients with a kidney biopsy, had a second relevant diagnosis in addition to MN: five (3%) patients had interstitial nephritis, five (3%) had a diabeticnephropathy was diagnosed and two (1%) had IgA nephropathy. These patients had a median eGFR of 51 mL/min/1.73 m 2 . Given the potential risks and costs of the biopsy procedure, the current recommendations in PLA2R-Ab positive patients [ 10 ] are to consider a kidney biopsy in the patients with unexplained deterioration of eGFR or unusual clinical presentation suggesting a secondary cause (in particular positive antinuclear antibodies), (Figure 4 ).
with endothelin-1 increased total b-catenin and phospho-NF-kB expression in wild-type glomeruli, but this effect was attenuated in Pod-ETRKO glomeruli. After streptozotocin injection to induce diabetes, wild-type mice developed mild diabeticnephropathy with microalbuminuria, mesangial matrix expansion, glomerular basement membrane thickening, and podocyte loss, whereas Pod-ETRKO mice presented less albuminuria and were completely protected from glomerulosclerosis and podocyte loss, even when uninephrectomized. Moreover, glomeruli from normal and diabetic Pod-ETRKO mice expressed substantially less total b-catenin and phospho-NF-kB compared with glomeruli from counterpart wild-type mice. This evidence suggests that endothelin-1 drives development of glomerulosclerosis and podocyte loss through direct activation of endo- thelin receptors and NF-kB and b-catenin pathways in podocytes. Notably, both the expression and function of the ETBR subtype were found to be important. Furthermore, these results indicate that activation of the endothelin-1 pathways selectively in podocytes mediates pathophysiologic crosstalk that inﬂuences mesangial architecture and sclerosis.
For several decades, Western countries have been facing an increasing incidence of allergies and autoimmune diseases. Immune-mediated conditions are thought to result from a complex interplay between genetic predisposition, immune dysregulation, and environmental factors ( 1 ). Membranous nephropathy (MN) is a rare autoimmune disease (incidence 1.3 cases per 100 000 inhabitants affecting more men than women) ( 2 ) with an increasing prevalence ( 3 ), characterized by subepithelial immune deposits containing IgG and complement fractions with alteration of the glomerular basement membrane structure ( 4 , 5 ) related to autoantibodies against podocyte proteins: M-type phospholipase A2 receptor 1 (PLA2R1) or thrombospondin type-1 domain-containing 7A (THSD7A) in 70% and 3% of patients, respectively ( 6 – 9 ). Anti-PLA2R1 and anti-THSD7A antibodies titers correlate with disease activity and predict disease outcome ( 10 , 11 ). Disease evolution is highly variable with spontaneous remission, persistent proteinuria or end-stage kidney diseases (ESKD) ( 12 ). Rituximab is a ﬁrst line anti-CD20 immunosuppressive drug often used to treat MN patients which induces remission in about 35% to 67% of cases ( 13 – 15 ), while MN recurs after remission in about 20% of cases ( 14 ). Nephrotic syndrome has been correlated with an exceptionally high risk of venous thromboembolic events ( 16 , 17 ).
The dual effect of pathogenic antibodies is well illustrated by anti-NEP antibodies. Mothers producing anti-NEP IgG1 transmitted the disease whereas those producing anti-NEP IgG4 only, failed to do so. These findings point to a critical role of the classical pathway of complement activation as attested by C1q deposition, whereas in adult cases of PLA2R-related autoimmune nephropathy anti-PLA2R IgG4 is prevailing and C1q deposits are rare. On the other hand, NEP also has important enzymatic in the kidney where it is present at the podocyte surface, in the brush border of the proximal tubule, and in vessel walls . NEP-specific IgG1 had a strong inhibitory effect, whereas IgG4 had a very weak effect. Inhibition of NEP activity by anti-NEP IgG1 antibodies may alter metabolism of a number of regulatory peptides and thus have an impact on glomerular hemodynamics, endothelial permeability, tubular function, and podocyte lesions [26, 31, 44, 45]. These alterations may be the cause of the unusual, severe ischemic lesions observed in the biopsy and of the disappearance of NEP from the brush border of proximal tubules, as also noted in ischemic and toxic acute kidney injuries [42, 52].
diabetic cardiomyopathy. In contrast, NOS2 and NOS 3 proteins were undetectable in cardio- myocytes [ 6 ].
Several points should be taken into consideration in interpreting our results. First, this study was conducted in rat myocardium, which differs from human myocardium. Second, we employed an animal model of type-1 diabetes and we studied only short term effects of diabe- tes and without insulin treatment. This model requires careful extrapolation to human diabe- tes, particularly type 2 diabetes although it is considered to be a well-established and reliable model for diabetes mellitus [ 44 ]. Third, analysis of gene expression using a microarray has its own limitations related to tissue selection, presentation and interpretation of vast amounts of data. Selected results were confirmed by Western blotting but not for all modified genes. Fourth, the dose of atorvastatin used in the study is higher than in human treatment but is the common regimen used in experimental studies [ 28 ]. The effects of other statin drugs have not been evaluated here. Since statins often exhibit class effects, some differences may be expected [ 45 ]. Lastly, some concerns have been raised about an increased risk of diabetes onset after high-dose statin treatment [ 46 ]. However the benefits of statins in overt diabetes have been widely studied and are not subject to controversy. In contrast, the benefit of statin in critically ill patients remained to be determined [ 47 – 48 ].
Factors associated, by univariate analysis, with wound improvement at the 1-month follow-up appointment and following standardized treatment. DFI, diabetic foot infection; SD, standard deviation; y, years. * Statistically significant.
principal component analysis of our culturomics approach, we observed that the presence of F. magna was associated with an unfavorable outcome in relation to ulcers at 1 month and, although not statistically significant, we identified strict anaerobic species in more patients with no improvement. However, the majority of the wounds analyzed in this study were superficial (PEDIS grade 2). Therefore, the prevalence of anaerobes reported here (32%) probably falls well below that of deeper wounds.
 Kline GA, Edwards A. Antepartum and intra-partum insulin management of type 1 and type 2 diabetic women: impact on clinically signiﬁcant neonatal hypoglycemia. Diabetes Res Clin Pract 2007;77:223–30.
 Lepercq J, Abbou H, Agostini C, Toubas F, Francoual C, Velho G, et al. A standardized protocol to achieve normoglycaemia during labour and delivery in women with type 1 diabetes. Diabetes Metab 2008;34:33–7.
 Schmeltz LR, DeSantis AJ, Thiyagarajan V, Schmidt K, O’Shea-Mahler E, Johnson D, et al. Reduction of surgical mortality and morbidity in diabetic patients undergoing cardiac surgery with a combined intravenous and subcutaneous insulin glucose management strategy. Diabetes Care 2007;30:823–8.
2 Hospices civils de Lyon , Lyon,FR
* Correspondence should be adressed to: Catherine Calzada <firstname.lastname@example.org >
Increased oxidative stress is associated with type-2 diabetes and related cardiovascular diseases but oxidative modification of LDL has been partially characterized. Our aim was to compare the lipid and fatty acid composition as well as the redox status of LDL from diabetic patients and healthy subjects. First, to ensure that isolation of LDL by sequential ultracentrifugation did not result in lipid modifications, lipid composition and peroxide content were determined in LDL isolated either by ultracentrifugation or fast-protein liquid chromatography. Both methods resulted in similar concentrations of lipids, fatty acids, hydroxy-octadecadienoic acid (HODE) and malondialdehyde (MDA). Then, LDL were isolated by ultracentrifugation from 8 type-2 diabetic patients and 8 control subjects. Compared to control LDL, diabetic LDL contained decreased cholesteryl esters and increased triglyceride
Thus, BCV could constitute an antiviral alternative in cases with UL97 mutations when FOS is contraindicated or in cases of FOS nephrotoxicity. Foscarnet nephropathy was initially described in the 1980s as a frequent complication in AIDS patients undergoing treatment for CMV infection . In vivo trisodium foscarnet crystals mixed with sodium calcium salt were first identified by infrared microscopy in the glomerular capillary lumens and tubules of AIDS patients . Importantly, isotonic saline infusion of 1.5 to 2.5 liters per day was demonstrated to reduce this renal toxicity by increasing FOS clearance and constituted the best preventive strategy . Nonetheless, renal failure is possible with FOS despite appropriate hydration. Cases of biopsy-proven FOS crystal precipitation in the transplantation field are relatively scarce and are summarized in Table 1. With the exception of one lung recipient, all patients were kidney transplant recipients. FOS nephropathy does not seem to appear during the first days of therapy but rather after several weeks of treatment. Glomerular crystallization seems to be associated with worse acute kidney injury than isolated tubular crystal- lization . At worst, FOS nephropathy led to kidney graft loss. Interestingly, FOS precipitation was also observed in the lungs, heart, pancreas, and gastrointestinal tract in two patients with severe systemic crystal dissemination [10, 11].
Obstructive nephropathy is a frequently encountered situation in newborns. In previous studies, the urinary peptidome has been analyzed for the identification of clinically useful biomarkers of obstructive nephropathy. However, the urinary proteome has not been explored yet and should allow additional insight into the pathophysiology of the disease. We have analyzed the urinary proteome of newborns (n=5/group) with obstructive nephropathy using label free quantitative nanoLC-MS/MS allowing the identification and quantification of 970 urinary proteins. We next focused on proteins exclusively regulated in severe obstructive nephropathy and identified Arginase 1 as a potential candidate molecule involved in the development of obstructive nephropathy, located at the crossroad of pro- and anti-fibrotic pathways. The reduced urinary abundance of Arginase 1 in obstructive nephropathy was verified in independent clinical samples using both Western blot and MRM analysis. These data were confirmed in situ in kidneys obtained from a mouse obstructive nephropathy model. In addition, we also observed increased expression of Arginase 2 and increased total arginase activity in obstructed mouse kidneys. mRNA expression analysis of the related arginase pathways indicated that the pro-fibrotic arginase-related pathway is activated during obstructive nephropathy. Taken together we have identified a new actor in the development of obstructive nephropathy in newborns using quantitative urinary proteomics and shown its involvement in an in vivo model of disease. The present study demonstrates the relevance of such a quantitative urinary proteomics approach with clinical samples for a better understanding of the pathophysiology and for the discovery of potential therapeutic targets.
Aristolochia clematitis L. seeds co-harvested with crops. Here we show that AAs are long-lived soil contaminants that enter wheat and maize plants by root uptake, with strong pH-dependence. Soil and crops from Serbian farms in areas endemic for A. clematitis were found to be extensively contaminated with AAs, with contamination strongly correlated with local incidence of BEN. The persistence of AAs as soil contaminants suggests that weed control for A. clematitis plants is needed to reduce the incidence of BEN and aristolochic acid nephropathy, and that systematic surveys of soil and crop AA levels would identify high-risk regions and it is imperative to research soil remediation methods.
thickness of the wall. We indeed observed that some TCs have a considerably thickened wall
that may reach more than 100 microns. Another limitation was that our sample was small
and comprised only type 2 diabetics with a high prevalence of proliferative diabetic
retinopathy, possibly reflecting a poor glycemic control and/or other population-related
Even in control rats, the gain in body weight was much lower in CR animals than in IF ones ( Figure 1 ). This coincided with lower mean values for the plasma insulin concentration, insulinogenic index, and HOMA index in IF control rats than in CR control rats examined at sacrifice after overnight starvation ( Table 4 ). Thus, for these three variables, the values recorded in IF control rats averaged 71 .6 ± 8 .6% (n = 15; P < 0.06) of the mean corresponding values found in CR control rats (100 .0 ± 11 .5%; n = 15). Since such distinctions between IF and CR control rats could not be ascribed to any di ﬀerence in either food intake or the responsiveness to D-glucose of isolated pancreatic islets incubated in vitro, they suggest a more stressful situation in CR control rats than in IF control animals. To a large extent, a comparable situation may prevail in CR as distinct from IF diabetic animals.