Journal of endocrinology and diabetes最新文献

Aims: To test whether circulating neurotoxic autoantibodies increase in adult type 2 diabetes mellitus with Parkinson's disease (PD) or dementia. To identify the G-protein coupled receptor on neuroblastoma cells mediating neural inhibitory effects in diabetic Parkinson's disease plasma autoantibodies. To determine the mechanism of accelerated neuroblastoma cell death and acute neurite retraction induced by diabetic Parkinson's disease and dementia autoantibodies.

Methods: Protein-A eluates from plasma of twelve older adult male diabetic patients having Parkinson's disease (n=10) or dementia (n=2), and eight age-matched control diabetic patients were tested for ability to cause accelerated N2A neuroblastoma cell death and acute neurite retraction. Specific antagonists of G protein coupled receptors belonging to the G alpha q subfamily of heterotrimetric G-proteins, the phospholipase C/inositol triphosphate/Ca2+ pathway, or the RhoA/Rho kinase pathway were tested for ability to block diabetic Parkinson's disease/dementia autoantibody-induced neurite retraction or N2A accelerated cell loss. Sequential Liposorber LA-15 dextran sulfate cellulose/protein-A affinity chromatography was used to obtain highly-purified fractions of diabetic Parkinson's disease autoantibodies.

Results: Mean accelerated neuroblastoma cell loss induced by diabetic Parkinson's disease or dementia autoantibodies significantly exceeded (P = 0.001) the level of N2A cell loss induced by an identical concentration of the diabetic autoantibodies in control patients without these two co-morbid neurodegenerative disorders. Co-incubation of diabetic Parkinson's disease and dementia autoantibodies with two-hundred nanomolar concentrations of M100907, a highly selective 5-HT2AR antagonist, completely prevented autoantibody-induced accelerated N2A cell loss and neurite retraction. A higher concentration (500 nM-10μM) of alpha-1 adrenergic, angiotensin II type 1, or endothelin A receptor antagonists did not substantially inhibit autoantibody-induced neuroblastoma cell death or prevent neurite retraction. Antagonists of the inositol triphosphate receptor (2-APB, 50μM), the intracellular calcium chelator (BAPTA-AM, 30 μM) and Y27632 (10 μM), a selective RhoA/Rho kinase inhibitor, each completely blocked acute neurite retraction induced by sixty nanomolar concentrations of diabetic Parkinson's disease autoantibodies. Co-incubation with 2-APB (1-2 μM) for 8 hours' prevented autoantibody-induced N2A cell loss. The highly-purified fraction obtained after Liposorber LA/protein-A affinity chromatography in hypertriglyceridemic diabetic dementia and Parkinson's disease plasmas had apparent MWs > 30 kD, and displayed enhanced N2A toxicity requiring substantially higher concentrations of 5-HT2AR antagonists (M100907, ketanserin, spiperone) to effectively neutralize.

Conclusion: These data suggest increased autoant

Introduction: Varying prevalence rates of type 2 diabetes have been observed in different parts of the southern state of Kerala, India which is in an advanced stage of epidemiologic transition. Social patterning is evident in diabetes and therefore it was decided to undertake a study on estimating the prevalence of diabetes and associated social determinants.

Methodology: The adopted local self administration unit of the Medical College which is also the field practice area with a population of 25,096 was taken for the study. All the households in the area were visited and the details regarding self reported diabetes was collected after obtaining informed consent and analysis done by multivariate logistic regression.

Result: The prevalence of self reported diabetes in this coastal area was found to be low at 7.4%. Type 2 diabetes was also found to occur significantly earlier among the respondents belonging to the below poverty line. Age above 40 years (OR 2 95% CI 1.5-2.7, p=.000), marital status (OR 1.9 95% CI 1.1-2.1, p=.006) presence of comorbidities (OR 635 95% CI 389-969, p=.000), more than 8 years of schooling (OR 0.64 95% CI 0.46-0.86, p=.004), living conditions as represented by presence of household source of drinking water(OR 1.4 95% CI 1.01-1.5) were found to be independent predictors. Though there was increasing trend of diabetes among the forward caste line families after backward logistic regression this disappeared leaving behind the proxy of socioeconomic status, household source of drinking water.

Conclusion: Though, the state of Kerala is in an advanced stage of epidemiologic transition, coastal areas are still in the earlier phases of transition with low prevalence of type 2 diabetes mellitus. Higher education and better living conditions are important social determinants of diabetes though further studies are necessary to delineate the impact of economic status and education.

Aims: To identify the G-protein coupled receptor(s) on neuroblastoma and endothelial cells which mediate neural- and endothelial cell-inhibitory effects in plasma autoantibodies from a subset of older type 2 diabetes with neurologic and vascular co-morbidity. To determine the mechanism(s) of neurite retraction induced by diabetic pathologies' auto antibodies.

Methods: Protein-A eluates from plasma of 11 diabetic patients having nephropathy, moderate-severe obesity and/or complications in which increased inflammation plays a role (depression, Parkinson's disease, atrial fibrillation, obstructive sleep apnea) were tested for neurite retraction and decreased survival in N2A neuroblastoma cells, and decreased survival in pulmonary artery endothelial cells. Specific antagonists of G protein coupled receptors belonging to the G alpha q subfamily of hetero trimetric G proteins or the phospholipase C/inositol triphosphate/Ca2+ pathway were tested for modulatory effects on diabetic pathologies' autoantibody-induced N2A neurite retraction, or cell survival.

Results: Co-incubation with specific antagonists of the 5-hydroxytryptamine- 2A receptor significantly prevented acute N2A neurite retraction induced by 50-100 nM concentrations of diabetic pathologies' autoantibodies. Protection against neurite retraction (M100907> spiperone> ketanserin) closely paralleled the antagonists' potency order at the 5-HT2-AR. Neuroblastoma or endothelial cell death (after 24 hours incubation) with 50-100 nM autoantibodies was completely or nearly completely (91%) prevented by co-incubation with 200 nM M100907, a highly selective 5-HT2-AR antagonist. Alpha-1 adrenergic, angiotensin II, metabotropic glutamate 5, or endothelin A (100 nM-10µM) receptor antagonists did not substantially inhibit autoantibody-induced cell death. The intracellular calcium chelator (BAPTA-AM, 50 µM) and inhibitors of the inositol triphosphate (IP3) receptor (2-APB, 50µM), and phospholipase C-gamma (U73144, 1µM) each significantly protected against autoantibody-induced acute N2A neurite retraction.

Conclusion: These data suggest that neural- and endothelial- inhibitory effects in autoantibodies from older adult diabetes with nephropathy and obesity/inflammation-associated complications are mediated by agonist autoantibodies directed against the 5-hydroxytryptamine 2 receptor positively coupled to the phospholipase C/inositol triphosphate/ cytosolic Ca2+ release pathway.

11 aim: Obstructive sleep apnea increases in diabetes and morbid obesity. We tested a hypothesis that circulating autoantibodies in adult type 2 diabetes which increase in association with morbid obesity are capable of causing long-lasting neuronal depolarization and altered calcium release in mouse atrial cardiomyocytes.

12 methods: Protein-A eluates from plasma of 14 diabetic obstructive sleep apnea patients and 17 age-matched diabetic patients without sleep apnea were tested for effects on depolarization and neurite out growth in N2a mouse neuroblastoma cells. The mechanism of autoantibody-mediated neurite outgrowth inhibition was investigated in co-incubation experiments of diabetic obstructive sleep apnea autoantibodies with specific antagonists of G-protein coupled receptors or the RhoA/Rho kinase signaling pathway. Following long-term storage of the protein-A eluates (to allow spontaneous proteolysis and IgG subunit dissociation), plasma autoantibodies from diabetic obstructive sleep apnea, cancer or control patients were compared for enhancement of inhibitory effects on endothelial cell survival. Size exclusion chromatography performed (in the presence or absence of a specific membrane type 1-matrix metalloproteinase inhibitor) was used to characterize the IgG autoantibody subunit(s) or fragments associated with peak neurotoxicity in diabetic obstructive sleep apnea.

13 results: Diabetic obstructive sleep apnea (n = 14) autoantibodies caused a significant increase (P = 0.01) in membrane depolarization in N2a mouse neuroblastoma cells compared to control diabetic patients (n = 15) not suffering with obstructive sleep apnea. Process extension in N2A mouse neuroblastoma cells was significantly inhibited (P = 0.01) by diabetic obstructive sleep apnea (n = 9) autoantibodies compared to effects from identical 10 μg/mL concentrations of control diabetic autoantibodies in patients without obstructive sleep apnea. Ten micromolar concentrations of SCH-202676, a G-protein coupled receptor antagonist (n = 5) or ten micromolar concentration of Y27632, a selective Rho kinase inhibitor (n = 6), each significantly prevented (P < 0.001) neurite outgrowth inhibition by diabetic obstructive sleep apnea autoantibodies. Autoantibodies in representative patients with obstructive sleep apnea and either atrial fibrillation or left ventricular hypertrophy evoked acute large increases in intracellular Ca²⁺ in HL-1 mouse atrial cardiomyocytes. The magnitude of intracellular Ca²⁺ release was dose-dependently significantly correlated to the electrocardiographic Cornell voltage-duration product. Gel filtration of diabetic obstructive sleep apnea autoantibodies revealed peak neurotoxicity associated with MWs corresponding to IgG light chain dimer(s), monomers or half-light chains as well as a novel ∼ 5.5 kD putative light chain fragment.

14 conclusions: <