Six-year follow-up of a cohort of 203 patients with diabetes after screening for silent myocardial ischaemia.

AIMS: To determine the prognosis of patients with Type 1 or Type 2 diabetes, 6 years after screening for silent myocardial ischaemia (SMI). METHODS: Two hundred and three asymptomatic patients with diabetes underwent systematic SMI screening. From the results of this screening, they were allocated to one of three groups: patients (n = 171) with negative screening; patients (n = 32) with positive screening; and patients (n = 21) with positive screening and coronary stenosis. Six years after the initial assessment, all patients were re-assessed. All events [death, cardiac death, non-fatal major cardiac events (NFMCEs)--acute myocardial infarction, ventricular rhythm disorders, heart failure, unstable angina] were recorded. RESULTS: Fifteen patients were lost to follow-up. Patients (n = 20) with positive SMI screening and coronary stenosis had a higher risk of NFMCEs (35% vs. 7%, P < 0.001), and a higher mortality rate (35% vs. 15%, P < 0.05) compared with patients (n = 157) with negative screening. SMI-positive patients (n = 31) had a higher NFMCE rate compared with negative SMI screening patients, although overall mortality rate was no different. Cancer was the leading cause of death (36.4%). In multivariate analysis, major cardiac events (cardiac death and NFMCE) were related to baseline age, body mass index and coronary stenosis (P < 0.01). CONCLUSIONS: Patients with diabetes and SMI have a very poor prognosis as assessed by cardiac events or death, especially in the presence of coronary stenosis.

C-peptide replacement improves weight gain and renal function in diabetic rats.

AIM: Recent experimental and clinical data suggest that C-peptide replacement during type 1 diabetes exerts beneficial effects on diabetic nephropathy. The aim of this study was to determine if physiological C-peptide administration in replacement dose during 28 days had beneficial effects on metabolic status and renal functions in type-1 diabetic rats. METHODS: Four groups of rats were investigated: a non diabetic group treated with buffer (C group, n=6), three streptozotocin diabetic-induced groups treated with either buffer (D group, n=6), insulin (D-I group, n=6) or rat homologous C-peptide (D-C group, n=6). Weight gain was measured every week. All animals were housed in metabolic cages on day 28 for assessment of metabolic data. Blood and urine samples were collected to allow measurement of plasmatic osmolality, C-peptide concentration, sodium, and glucose losses and proteinuria. Glomerular filtration rate (GFR) was determined by creatinine clearance. RESULTS: All streptozotocin-treated animals were diabetic. Glycaemic control (mg/dl), was markedly improved in D-I (133+/-65) when compared with either D (547+/-49, P<0.05) or D-C (520+/-48, P<0.05) groups. Conversely, weight gain during the study, was improved in D-I and D-C as compared with D animals (135+/-13 and 41+/-18 vs 18+/-21 respectively), despite different glycaemic control. Diabetes-induced glomerular hyperfiltration (ml/min/kg), urinary protein leakage (g/kg/day), and Na urinary losses (mmol/100 g/day) respectively, were significantly (P<0.05) reduced in D-C (3.95+/-0.6; 0.08+/-0.06; 1.5+/-0.9) in comparison with D (4.95+/-0.8; 0.18+/-0.16; 3.7+/-2.1) and D-I (5+/-0.9; 0.19+/-0.11; 2.7+/-0.8) animals. Plasmatic osmolality was significantly increased in D group whereas there were no differences between C group and D-C group. Food and water intakes, urinary volume as well as urinary glucose losses were not significantly different between D-C and D groups. CONCLUSIONS: C-peptide administration in replacement dose to streptozotocin diabetic rats induces weight gain regardless hyperglycaemia or glycosuria. Diabetic animals supplemented with C-peptide exhibit better renal function resulting in reduced urinary sodium waste and protein excretion together with reduction of the diabetes-induced glomerular hyperfiltration.

Factors related to CSII compliance.

Many studies have demonstrated the metabolic efficacy of continuous subcutaneous insulin infusion (CSII) and particularly a reduction of glycaemic fluctuations in type 1 diabetic patients. Despite this benefit, many patients decide to discontinue the use of CSII. To determine the factors related to discontinuation of CSII we analyzed clinical data from a group of 70 patients who had been consecutively started on this treatment from April 2000 to April 2002. Patients were followed for up to 2 years. Eighteen (25.7%) patients decided to terminate CSII during the study after an average of 235 days (range 21-293). The reasons for stopping CSII were decision of the patients (10), end of pregnancy (4), needle site infections (3) and lack of compliance (1). No significant difference was found between patients who had continued and those who had discontinued CSII for age, duration of diabetes, reasons for starting CSII, marital status, prepump concentration of HbA1c and prepump frequency of hypoglycaemia. There tend to be more discontinuations for pregnant women, patients attending hospital visits versus liberal practitioner and patients with lower educational level (below or over baccalaureat) although none of these differences was statistically significant. In conclusion we could not identify any predictive factor of CSII discontinuation.

Insulin detemir used in basal-bolus therapy in people with type 1 diabetes is associated with a lower risk of nocturnal hypoglycaemia and less weight gain over 12 months in comparison to NPH insulin.

AIM: The aim of this study was to compare the long-term safety and efficacy of twice-daily insulin detemir or NPH insulin as the basal component of basal-bolus therapy in people with type 1 diabetes. METHODS: A multicentre, open-label, parallel-group study was conducted over 12 months and completed by 308 people (from an original randomized cohort of 428). Patients were randomized in a 2:1 ratio to receive insulin detemir or NPH insulin before breakfast and dinner, with insulin aspart at mealtimes. RESULTS: Glycaemic control improved in both groups with HbA(1c) decreasing by 0.64 and 0.56% point in the insulin detemir and NPH insulin groups, reaching baseline-adjusted final values of 7.53 +/- 0.10% and 7.59 +/- 0.13%, respectively. No significant difference was apparent between treatments in terms of HbA(1c), fasting plasma glucose or 9-point blood glucose profiles. Fewer hypoglycaemic events (major and minor) occurred in association with insulin detemir compared with NPH insulin, but the overall hypoglycaemic risk did not differ statistically significantly (RR for detemir, 0.78 [0.56-1.08]). However, the risk of nocturnal hypoglycaemia during the maintenance phase (month 2-12) was 32% lower in the detemir group (p = 0.02) and lower in every month. This risk reduction remained statistically significant after correction for HbA(1c). After 12 months, baseline-adjusted mean body weight was significantly lower in the insulin detemir group than in the NPH insulin group (p < 0.001). CONCLUSIONS: In long-term basal-bolus therapy, insulin detemir with insulin aspart as mealtime insulin is well tolerated and reduces the risks of nocturnal hypoglycaemia and weight gain compared to NPH insulin.

C-peptide, Na+,K(+)-ATPase, and diabetes.

Na+,K(+)-ATPase is an ubiquitous membrane enzyme that allows the extrusion of three sodium ions from the cell and two potassium ions from the extracellular fluid. Its activity is decreased in many tissues of streptozotocin-induced diabetic animals. This impairment could be at least partly responsible for the development of diabetic complications. Na+,K(+)-ATPase activity is decreased in the red blood cell membranes of type 1 diabetic individuals, irrespective of the degree of diabetic control. It is less impaired or even normal in those of type 2 diabetic patients. The authors have shown that in the red blood cells of type 2 diabetic patients, Na+,K(+)-ATPase activity was strongly related to blood C-peptide levels in non-insulin-treated patients (in whom C-peptide concentration reflects that of insulin) as well as in insulin-treated patients. Furthermore, a gene-environment relationship has been observed. The alpha-1 isoform of the enzyme predominant in red blood cells and nerve tissue is encoded by the ATP1A1 gene. A polymorphism in the intron 1 of this gene is associated with lower enzyme activity in patients with C-peptide deficiency either with type 1 or type 2 diabetes, but not in normal individuals. There are several lines of evidence for a low C-peptide level being responsible for low Na+,K(+)-ATPase activity in the red blood cells. Short-term C-peptide infusion to type 1 diabetic patients restores normal Na+,K(+)-ATPase activity. Islet transplantation, which restores endogenous C-peptide secretion, enhances Na+,K(+)-ATPase activity proportionally to the rise in C-peptide. This C-peptide effect is not indirect. In fact, incubation of diabetic red blood cells with C-peptide at physiological concentration leads to an increase of Na+,K(+)-ATPase activity. In isolated proximal tubules of rats or in the medullary thick ascending limb of the kidney, C-peptide stimulates in a dose-dependent manner Na+,K(+)-ATPase activity. This impairment in Na+,K(+)-ATPase activity, mainly secondary to the lack of C-peptide, plays probably a role in the development of diabetic complications. Arguments have been developed showing that the diabetes-induced decrease in Na+,K(+)-ATPase activity compromises microvascular blood flow by two mechanisms: by affecting microvascular regulation and by decreasing red blood cell deformability, which leads to an increase in blood viscosity. C-peptide infusion restores red blood cell deformability and microvascular blood flow concomitantly with Na+,K(+)-ATPase activity. The defect in ATPase is strongly related to diabetic neuropathy. Patients with neuropathy have lower ATPase activity than those without. The diabetes-induced impairment in Na+,K(+)-ATPase activity is identical in red blood cells and neural tissue. Red blood cell ATPase activity is related to nerve conduction velocity in the peroneal and the tibial nerve of diabetic patients. C-peptide infusion to diabetic rats increases endoneural ATPase activity in rat. Because the defect in Na+,K(+)-ATPase activity is also probably involved in the development of diabetic nephropathy and cardiomyopathy, physiological C-peptide infusion could be beneficial for the prevention of diabetic complications.

The Medi-RIVAGE study (Mediterranean Diet, Cardiovascular Risks and Gene Polymorphisms): rationale, recruitment, design, dietary intervention and baseline characteristics of participants.

OBJECTIVE: To report the rationale, recruitment, design, dietary intervention and baseline characteristics of participants in the Medi-RIVAGE study (Mediterranean Diet, Cardiovascular Risks and Gene Polymorphisms). DESIGN: A randomised, parallel trial comparing a new nutritional programme with a conventional programme. SETTING: Centre for Detection and Prevention of Arteriosclerosis, Timone University Hospital, Marseille, France, and collaborating teams. SUBJECTS: Two hundred and twelve male and female volunteers with at least one cardiovascular risk factor. INTERVENTION: A Mediterranean-type diet characterised mainly by the quality of fatty acids, amount of fish, vegetable foodstuffs and fibre was proposed and compared with a usually prescribed, low-fat/cholesterol diet. Body mass index, fasting lipids and lipoproteins, apolipoproteins, glucose, insulin and homocysteine were the main outcome measures. Gene polymorphisms of interest were determined. RESULTS: Characteristics of men in the two arms were comparable with regard to sociodemographic variables, and clinical and biological cardiovascular risk factors. There were few differences between the groups of women (cholesterol-related parameters, P<0.05). There was no difference between arms in allelic distribution of the gene polymorphisms studied. Saturated fat and protein intakes were high while carbohydrate and fibre intakes were low, but with no difference between arms. Overall, the nutritional markers were comparable in both arms with few exceptions. Correlations between nutritional intakes and plasma nutrient levels ranged from 0.19 (beta-carotene) to 0.47 (folate). CONCLUSIONS: The comparability of the two arms is notable and warrants a low risk of biases. Current diet departs from the traditional Mediterranean one. The assessment of nutritional intake is validated by correlations obtained between dietary intake and relevant biomarkers. This will be important to estimate participant compliance and to analyse intervention data.

[Genetics of diabetic complications: peripheral neuropathy]. / Génétique des complications du diabète: neuropathie périphérique.

Diabetic neuropathy is a common complication of type 1 and 2 diabetes mellitus. Chronic hyperglycaemia and/or insulin deficiency in the peripheral nerve lead to metabolic and vascular disturbances, responsible for the functional alterations and the characteristic histological abnormalities observed in the nerve fibre. Recently, genetic factors have been described, suggestive of a predisposition and/or a protective effect for diabetic neuropathy in certain patients. The search for these genetic factors through the study of polymorphism of gene involved in the various metabolic and vascular pathways, is currently increasing, but with contradictory results. The main studies and data are reviewed in this Article. The identification of candidate-genes should allowed, in the future, to better identify and manage diabetic patients at-risk for peripheral neuropathy.

How and when to use an alternative site in self-monitoring of blood glucose.

Taking care of diabetic patients has considerably been improved for approximately fifty years both in the therapeutic field and in the glycaemia monitoring field. Prospective studies conducted on large cohorts have clearly shown the importance of taking optimal care of such patients in order to prevent the occurrence or aggravation of chronic diabetes-associated complications. However, despite the simplification of self-monitoring of blood glucose through technological developments, drawbacks, some of which are linked to the sampling site, the fingertip, still slow down the patients' compliance. The use of an alternative site seems to be one of the solutions to offer in order to improve their monitoring and hence, their metabolic control. The development of such a monitoring mode has been slowed down after revealing, in some studies, a probably physiological delay in the detection of glycaemia variations, at the level of the alternative sites. Despite such conflicting observations, interest of using such alternative sites is to be defined in self-monitoring. Beside fast glycaemia variations, it proved to be reliable.

Peripheral diabetic neuropathy and polyunsaturated fatty acid supplementations: natural sources or biotechnological needs?

The two essential fatty acids linoleic and alpha-linolenic acids, precursors of the n-6 and n-3 PUFA family, respectively, are known to play a strong regulatory function on cells via their incorporation into membrane phospholipids, and also on microcirculation by the production of eicosanoids. Moreover, diabetes mellitus induces impairment in PUFA metabolism due to an inhibition of desaturases, the enzymes involved in their synthesis. The decrease in PUFA bioavailability will conduct to marked alterations in membranes as well as impairment of the microcirculation. Those metabolic perturbations are involved in part in the degenerative complications of diabetes such as neuropathy. Nutritional supplementations with PUFA have given very interesting results in experimental diabetic neuropathy but also in human diabetic neuropathy. The gamma linolenic and arachidonic acids, members of the n-6 family, prevent the physiological abnormalities associated to neuropathy. The results obtained with the n-3 family PUFA are more discordant, probably because of the simultaneous use of eicosapentaenoic and docosahexaenoic acids. Nevertheless, the use of docosahexaenoic acid-enriched phospholipids produced positive results in the treatment of experimental diabetic neuropathy. These PUFA are available from natural sources but a biotechnological demand exists to provide these PUFA in different structural forms.

Reproducibility of plasma insulin kinetics during intraperitoneal insulin treatment by programmable pumps.

OBJECTIVE: To study the reproducibility of plasma insulin kinetics during intraperitoneal (IP) insulin therapy using an implanted programmable pump in patients with type 1, insulin dependent, diabetes mellitus (IDDM). RESEARCH DESIGN AND METHODS: In a group of ten type 1 IDDM patients beginning chronic IP insulin treatment with an implanted pump, plasma free insulin profiles were determined from 12: 00 am to 12: 30 pm on two separate test days, one month apart. Anti-insulin antibody (AIA) levels were measured on each test day. RESULTS: From test day 1 to test day 2, no difference was observed in morning fasting free insulin levels (m +/- SD): 9.7 +/- 5.4 mU/L versus 9.8 +/- 5.3 mU/L, insulin peak values: 19.1 +/- 17 mU/L versus 20.8 +/- 9.9 mU/L, time to peak: 40 +/- 15 versus 42.8 +/- 16 minutes or post-bolus area under the plasma free insulin curve (AUC): 40.7 +/- 29 mU/L.h versus 45.5 +/- 29 mU/L.h. The intrapatient coefficient of variation was 14.4 +/- 13% for insulin peaks and 16.9 +/- 9.2% for post-bolus AUC. A significant increase in AIA levels (m +/- SD) was observed from 16.5 +/- 18% on test day 1 to 28.1 +/- 28% on test day 2. CONCLUSIONS: The reproducibility of plasma free insulin profiles is highly satisfactory during insulin delivery by the IP route using an implanted device.

[History, accuracy and precision of SMBG devices]. / Technologie et fiabilité de l'autosurveillance glycémique: historique et état actuel.

Self-monitoring of blood glucose started only fifty years ago. Until then metabolic control was evaluated by means of qualitative urinary blood measure often of poor reliability. Reagent strips were the first semi quantitative tests to monitor blood glucose, and in the late seventies meters were launched on the market. Initially the use of such devices was intended for medical staff, but thanks to handiness improvement they became more and more adequate to patients and are now a necessary tool for self-blood glucose monitoring. The advanced technologies allow to develop photometric measurements but also more recently electrochemical one. In the nineties, improvements were made mainly in meters' miniaturisation, reduction of reaction time and reading, simplification of blood sampling and capillary blood laying. Although accuracy and precision concern was in the heart of considerations at the beginning of self-blood glucose monitoring, the recommendations of societies of diabetology came up in the late eighties. Now, the French drug agency: AFSSAPS asks for a control of meter before any launching on the market. According to recent publications very few meters meet reliability criteria set up by societies of diabetology in the late nineties. Finally because devices may be handled by numerous persons in hospitals, meters use as possible source of nosocomial infections have been recently questioned and is subject to very strict guidelines published by AFSSAPS.

Differential effect of omega3 PUFA supplementations on Na,K-ATPase and Mg-ATPase activities: possible role of the membrane omega6/omega3 ratio.

Several functional properties of Na,K-ATPase are strongly dependent on membrane fatty acid composition, but the underlying mechanism is still not well defined. We have studied the effects of two types of supplementations enriched in the w3 polyunsaturated fatty acids on the Na,K-ATPase and Mg-ATPase activities in sciatic nerve (SN) and red blood cells (RBC). Eight groups of rats, controls and diabetics, received a standard diet, supplemented or not with 30 or 60 mg/kg/day of docosahexaenoic acid (DHA) or with soybean for eight weeks. Diabetes induced significant decrease of Na,K-ATPase activity in SN (-23%) and RBC (-25%), without affecting Mg-ATPase activity. In RBC, soybean and DHA supplementations caused significant increases in Na,K-ATPase activity (in various range, +13% to +145%) in all groups, and in Mg-ATPase activity in control soybean (+65%), control and diabetic DHA high dose (+39%, +53%) and diabetic DHA low dose (+131%) groups. In SN, the soybean caused a significant decrease in Na,K-ATPase activity (-26%) and still more in the diabetic group (-53%). The DHA diet induced a slight decrease in activity in control groups, whilst during diabetes, at high dose, we noted an aggravation of this decrease (-36%). Mg-ATPase activity was not modified by supplementations except for the low dose of DHA where the activity was slightly decreased in the control group (-16%). The supplementations induced multiple tissue-specific modifications in the membrane fatty acid composition of RBC and of SN homogenates. Several specific correlations have been found between variations in fatty acids amounts and Na,K-ATPase activity in these tissues but only in RBC for Mg-ATPase activity. Indeed, we observed that the variations in Na,K-ATPase activity are positively and significantly correlated with changes in the omega6/omega3 ratio in SN as well as in RBC. These data clearly show, for the first time, that the diet could modulate the Na,K-ATPase activity via the omega6/omega3 ratio in the membranes. A similar correlation was observed with Mg-ATPase activity in RBC, suggesting also a dietary regulation of the enzyme; but for the SN, this activity might be regulated by a different omega6/omega3 ratio or by another pathway.

Genetic and environmental regulation of Na/K adenosine triphosphatase activity in diabetic patients.

Even if the pathogenesis of diabetic neuropathy is incompletely understood, an impaired Na/K adenosine triphosphatase (ATPase) activity has been involved in this pathogenesis. We previously showed that a restriction fragment length polymorphism (RFLP) of the ATP1-A1 gene encoding for the Na/K ATPase's alpha 1 isoform is associated with a low Na/K ATPase activity in the red blood cells (RBCs) of type 1 diabetic patients. We thus suggested that the presence of the variant of the ATP1A1 gene is a predisposing factor for diabetic neuropathy, with a 6.5% relative risk. Furthermore, there is experimental evidence showing that lack of C-peptide impairs Na/K ATPase activity, and that this activity is positively correlated with C-peptide level. The aim of this study was to evaluate the respective influence of genetic (ATP1-A1 polymorphism) and environmental (lack of C-peptide) factors on RBC's Na/K ATPase activity. Healthy and diabetic European and North African subjects were studied. North Africans were studied because there is a high prevalence and severity of neuropathy in this diabetic population, and ethnic differences in RBC's Na/K ATPase activity are described. In Europeans, Na/K ATPase activity was significantly lower in type 1 (285 +/- 8 nmol Pi/mg protein/h) than in type 2 diabetic patients (335 +/- 13 nmol Pi/mg protein/h) or healthy subjects (395 +/- 9 nmol Pi/mg protein/h). Among type 2 diabetic patients, there was a significant correlation between RBC's Na/K ATPase activity and fasting plasma C-peptide level (r = 0.32, P <.05). In North Africans, we confirm the ethnic RBC's Na/K ATPase activity decrease in healthy subjects (296 +/- 26 v 395 +/- 9 nmol Pi/mg protein/h, r < 0.05), as well as in type 1 diabetic patients (246 +/- 20 v 285 +/- 8 nmol Pi/mg protein/h; P <.05). However, there is no relationship between the ATP1A1 gene polymorphism and Na/K ATPase activity. ATP1A1 gene polymorphism could not explain the ethnic difference. We previously showed that Na/K ATPase activity is higher in type 1 diabetic patients without the restriction site on ATP1A1 than in those heterozygous for the restriction site. This fact was not observed in healthy subjects. In type 2 diabetic patients, association between ATP1A1 gene polymorphism and decreased enzyme activity was found only in patients with a low C-peptide level. Therefore, the ATP1-A1 gene polymorphism influences Na/K ATPase activity only in case of complete or partial C-peptide deficiency, as observed in type 1 and some type 2 diabetic patients, without any correlation with hemoglobin A1c (HbA1c). Correlation observed between C-peptide levels and RBC's Na/K ATPase suggests that the deleterious effect of C peptide deficiency on Na/K ATPase activity is worse in the presence of the restriction site. This may explain the high relative risk of developing the neuropathy observed in type 1 diabetic patients bearing the variant allele.

Gamma-linolenic acid restores renal medullary thick ascending limb Na(+),K(+)-ATPase activity in diabetic rats.

In diabetes, the activity of Delta-6 desaturase, which converts linoleic acid (LA) into gamma-linolenic acid (GLA), the first step of arachidonic acid (AA) synthesis, is decreased, leading to alterations in membrane phospholipid composition. On the other hand, 12 wk after the onset of diabetes, Na(+),K(+)-ATPase activity is reduced in many organs, including the kidney. The medullary thick ascending limb (MTAL) reduced Na(+),K(+)-ATPase activity, whereas the sodium load secondary to glomerular hyperfiltration was increased. The aim of our study was to examine whether the changes in membrane fatty acid composition resulting from the inhibition of Delta-6 desaturase may be involved in the decreased Na(+),K(+)-ATPase activity observed in the outer MTAL after 12 wk of diabetes. GLA is a fatty acid that by-passes the Delta-6 desaturase step. We measured the membrane fatty acid composition and the Na(+),K(+)-ATPase activity in the renal outer medulla of control and streptozotocin (STZ)-induced diabetic rats 12 wk after the induction of diabetes. Measurements were performed after supplementation of control rats with sunflower oil (SO) or GLA for 12 wk, and supplementation of 12 wk diabetic rats with SO for 12 wk or with GLA for 6 or 12 wk. Supplementation with GLA not only prevented the decrease in Na(+),K(+)-ATPase activity observed after 12 wk of diabetes but also time dependently stimulated Na(+),K(+)-ATPase activity in the outer medulla. The changes in Na(+),K(+)-ATPase activity were related to parallel changes in the amount of Na(+),K(+)-ATPase alpha(1) subunit protein. In addition, in diabetic rats only, Na(+),K(+)-ATPase activity was positively correlated with the amount of AA present in cell membranes (r = 0.92, P < 0.05). Our results indicate that nutritional GLA supplementation increases Na(+),K(+)-ATPase activity and expression in diabetic rats. In addition, the positive correlation between AA content and Na(+),K(+)-ATPase activity suggests that in diabetic rats, alterations in membrane fatty acid composition contribute to the decreased Na(+),K(+)-ATPase activity in outer medulla.

Evidence of time-dependent changes in renal medullary Na,K-ATPase activity and expression in diabetic rats.

The medullary thick ascending limb (MTAL) of the kidney displays structural changes during long term diabetes. After twelve weeks of diabetes, there is controversy over the changes in Na,K-ATPase activity. To observe the long-term changes, we studied MTAL Na,K-ATPase activity and protein expression in diabetic animals 6 (6W) and 12 weeks (12W) after induction of diabetes with streptozotocin. Three groups were studied, one control group, one group 6W after, and one group 12W after induction of diabetes. Membrane fractions from the inner strip of the outer medulla representing MTAL were isolated. Na,K-ATPase activity and western blottings of alpha1- and beta1-subunits were carried out. 6W diabetes resulted in an increase, and 12W in a decrease in the MTAL Na,K-ATPase activity versus the control group (respectively 63.3 +/- 21.2; 7.5 +/- 2.4 and 31.6 +/- 11.4; micromol Pi/mg prot/hr +/- SEM). The Na,K-ATPase subunit expression was increased at 6W, and decreased after 12W, resulting in amounts below control values for both alpha1- and beta1-subunits. Our results confirm a diabetes-induced biphasic time-dependent alteration MTAL Na,K-ATPase activity, supported by similar changes in alpha1 and beta1 Na,K-ATPase subunits-expression.

Na,K-atpase alterations in diabetic rats: relationship with lipid metabolism and nerve physiological parameters.

Type 1 diabetes induces several metabolic and biochemical disturbances which result in the alteration ofNa,K-ATPase, an enzyme implicated in the physiopathology of neuropathy Several fatty acid supplementations lessen this alteration. The aims of this study were to determine the possible relationships between Na,K-ATPase activity in nerves and red blood cells (RBCs) and, on one hand, the fatty acid alterations induced by diabetes in these tissues and plasma and on the other, on nerve physiological parameters. Two groups of rats, control and diabetic (n = 15), were sacrified 8 weeks after induction of diabetes with streptozotocin. Nerve conduction velocity (NCV), nerve blood flow (NBF), Na,K-ATPase activity and membrane fatty acid composition of sciatic nerves, red blood cells (RBCs) and plasma were measured. NCV, NBF and Na,K-ATPase activity in RBCs and in sciatic nerves were significantly decreased in diabetic rats. We revealed a positive correlation between Na,K-ATPase activity in sciatic nerves and both NBF and NCV and between Na,K-ATPase activity in RBCs and NBF and the same activity in sciatic nerve. Diabetes induced major changes in plasma fatty acids and RBC membranes and less important changes in sciatic nerve membranes. Na,K-ATPase activity correlated negatively with C20: 4 (n-6) and C22: 4 (n-6) levels in nerves and with C18: 2 (n-6) levels in RBCs. During diabetes, changes in the membrane fatty acid composition suggest the existence of a tissue-specific regulation, and the decrease in Na,K-ATPase activity correlates with the alteration in the level of specific fatty acids in RBCs and sciatic nerves. Modifications in the lipidic environment of Na,K-ATPase would be involved in the alteration of its activity. Na,K-ATPase activity seems to be implicated in the decrease of both NCV and NBF during diabetes.

Accumulation of triglyceride-rich lipoprotein in subjects with abdominal obesity: the biguanides and the prevention of the risk of obesity (BIGPRO) 1 study.

The present study represents a new insight into the Biguanides and the Prevention of the Risk of Obesity (BIGPRO) 1 study population at inclusion. This population, selected basically on the basis of a high waist-to-hip ratio (>/=0.95 for men and >/=0.80 for women), is supposed to represent a group of patients with insulin resistance. The present study was undergone to establish whether apolipoprotein C-III (apoC-III) and apolipoprotein E (apoE) associated with apo B (apoC-III LpB and apoE LpB, respectively), considered to be markers of remnant accumulation, play a role in the hypertriglyceridemia associated with insulin resistance and whether they are related to other biological abnormalities frequently observed in this syndrome. In this population, the concentration of the markers of remnant accumulation increases with triglyceride levels. Therefore, correlation studies were realized to assess the relative effect of insulin and the markers of remnant accumulation on triglyceride plasma level. As a first attempt, a simple correlation analysis revealed that insulin is positively related to the markers of remnant accumulation only in hypertriglyceridemic patients (triglycerides >/=1.7 mmol/L). To assess the independent contribution of these markers, insulin, and other parameters related to the plasma triglyceride concentration, a stepwise multiple regression analysis was run. Results revealed that insulin and the markers of remnant accumulation (specifically, apoE LpB) are independent contributors to the plasma triglyceride concentration. Markers of the endothelial damage, plasminogen activator inhibitor-1, tissue plasminogen activator, and von Willebrand factor, which are often increased in the case of insulin resistance, were tested for their correlation with the markers of remnant accumulation. Plasminogen activator inhibitor-1 is positively correlated with these markers only in hypertriglyceridemic male subjects. It is concluded that increased insulin levels found in insulin resistance syndrome are associated with an increased production of triglyceride-rich lipoproteins enriched in apoC-III and apoE. The accumulation of these remnants and/or their abnormal composition in apoC-III and apoE could be an explanation for the development of hypertriglyceridemia in this syndrome.

The effects ex vivo and in vitro of insulin and C-peptide on Na/K adenosine triphosphatase activity in red blood cell membranes of type 1 diabetic patients.

The decrease in Na/K adenosine triphosphatase (ATPase) activity observed in several tissues of type 1 diabetic patients is thought to play a role in the development of long-term complications. Infusion of insulin may restore this enzyme activity in red blood cells (RBCs), and recent arguments have been developed for a similar role of C-peptide. The aims of this study were to determine whether insulin acts directly on the RBC enzyme and to evaluate the effect of C-peptide on Na/K ATPase activity. Thirty-nine C-peptide-negative type 1 diabetic patients were studied (blood glucose, 11.2 +/- 1.49 mmol/L; hemoglobin A1c [HbA1c], 8.9% +/- 0.1%, mean +/- SEM). Blood samples were obtained in the morning, before breakfast and insulin injection. Intact and living RBCs were resuspended in their own plasma and incubated with or without insulin (50 microU/mL) or C-peptide (6 nmol/L). Ex vivo by microcalorimetry, the heat produced after 1 hour by the enzyme-induced hydrolysis of adenosine triphosphate (ATP), was measured in a thermostated microcalorimeter at 37 degrees C. The results showed that Na/K ATPase activity was significantly increased by insulin (12.4 +/- 0.5 v 15.4 +/- 0.9 mW/L RBCs, P < .05, n = 23) but not by C-peptide (11.9 +/- 0.7 v 12.9 +/- 0.9 mW/L RBCs, NS, P = .26, n = 12). In another experiment, RBC suspensions were incubated at 37 degrees C in a water bath with or without insulin (50 microU/mL) or C-peptide (6 nmol/L) for 10 minutes. RBC membranes were isolated and Na/K ATPase activity was assessed by measuring inorganic phosphate release at saturating concentrations of all substrates. The results showed that insulin and C-peptide significantly increased RBC Na/K ATPase activity (342 +/- 25, P < .005 and 363 +/- 30, P < .005, respectively v255 +/- 22 nmol Pi x mg protein(-1) x h(-1), n = 14). We conclude that insulin and C-peptide act directly on RBC Na/K ATPase, thus restoring this activity in type 1 diabetic patients. The stimulatory effect of C-peptide observed in vitro on RBC Na/K ATPase activity confirms that C-peptide plays a physiological role.