The relationship of oxidative stress to thrombotic tendency in type 1 diabetic patients with retinopathy.

Increased free radical activity may contribute to thrombosis via effects on platelet aggregation and the prostanoid balance. To investigate this further we studied 15 Type 1 diabetic patients with retinopathy, matched with uncomplicated Type 1 patients for age, duration of diabetes and HbA1, together with matched healthy non-diabetic control subjects. The oxidative effects of free radicals as total diene conjugates and lipid peroxides were measured, together with redox status extracellularly as plasma albumin-thiols and intracellularly as erythrocyte superoxide dismutase activity. Platelet count, aggregation of platelets in whole blood to collagen, thromboxane B2, and prostacyclin stimulating factor (PGI2SF) were also assessed. Free radicals measured as lipid peroxides were significantly higher (9.6 (8.1-11.6) mumol l-1 (median and interquartile range) in diabetic patients with retinopathy than in control subjects (8.1 (7.4-9.2) mumol l-1; p less than 0.05). There were also significant reductions in redox status both extracellularly as plasma albumin thiols (408 (383-473) vs 490 (456-517) mumol l-1, p less than 0.001) and intracellularly as erythrocyte superoxide dismutase activity (34 (27-41) vs 44 (36-51) g l-1, p less than 0.05) between patients with retinopathy and control subjects. Platelet counts were increased in diabetic patients with retinopathy (p less than 0.05), as was collagen-induced platelet aggregation (p less than 0.01). Prostacyclin stimulating factor was reduced in patients with retinopathy (p less than 0.05) and correlated within the plasma with lipid peroxides (r = -0.53, p less than 0.04) and albumin thiols (r = 0.64, p less than 0.01). The results suggest that diabetic patients, particularly with retinopathy, are under oxidative stress and have an increased thrombotic tendency with increased platelet reactivity and a reduction in prostacyclin stimulating factor.     

Free radical activity in type 2 diabetes

Free radical activity has been implicated in the development of diabetic vascular complications in Type 1 diabetes. The aim of the present study was to investigate the levels of free radical scavengers, particularly erythrocyte superoxide dismutase, plasma and erythrocyte lysate thiol, and caeruloplasmin in 22 Type 2 diabetic patients clinically free of complications, and 15 comparable non-diabetic control subjects. The concentration (median (range] of both superoxide dismutase (23 (10-39) vs 45 (25-75) mumol l-1; p less than 0.001) and plasma thiol (374 (172-523) vs 460 (386-595) mumol l-1; p less than 0.01) were reduced in the diabetic group. There were no significant differences in the concentration of erythrocyte lysate thiol (199 (114-520) vs 188 (114-328) mumol l-1) or plasma caeruloplasmin (18 (9-31) vs 24 (6-50) mumol l-1) between the groups. This reduction in superoxide dismutase and the imbalance in the redox status of the plasma and lysate thiol demonstrated is consistent with an increase in free radical activity in Type 2 diabetes.     

Metabolic control may influence the increased superoxide generation in diabetic serum.

Superoxide anion (O2-) generation in serum from 10 Type 1 diabetic patients and 10 normal subjects was measured ex vivo. The amount of O2- production was significantly increased in diabetic serum 0.41 +/- 0.04 (+/- SD) vs 0.14 +/- 0.04 mumol l-1 min-1, p less than 0.001) and correlated with fasting plasma glucose and glycosylated protein levels in both diabetic (r = 0.72, p less than 0.01, and r = 0.62, p less than 0.05) and normal r = 0.75, p less than 0.01 and r = 0.64, p less than 0.05) subjects. Improved metabolic control in the diabetic patients was associated with a reduction of serum O2- production (0.28 +/- 0.06 mumol l-1 min-1, p less than 0.01), but the correlation between O2- levels and fasting plasma glucose and glycosylated protein concentrations was retained (r = 0.86 and r = 0.72, respectively, both p less than 0.01).     

The effect of ambient temperature on glucose tolerance.

Standard (75 g) oral glucose tolerance tests were performed at two different ambient temperatures (23 and 33 degrees C) in random order in 16 (eight obese) diabetic and 16 (eight obese) non-diabetic Nigerian subjects. Consistently higher plasma glucose values were found 120 min post-glucose ingestion at 33 degrees C, with mean differences of 0.5 (SE 0.3) and 4.5 (SE 1.5) mmol l-1 in the non-diabetic and diabetic subjects, respectively. This caused reclassification of two of the non-diabetic subjects as Impaired Glucose Tolerance at 33 degrees C, applying the WHO criteria. The difference was consistently greater (p less than 0.01) in the non-obese subjects (non-diabetic 0.8 (0.4), diabetic 6.9 (2.8) mmol l-1) than in the obese (non-diabetic 0.2 (0.4), diabetic 2.1 (0.9) mmol l-1). In the diabetic subjects, a negative correlation (r = -0.50, p less than 0.01) was established between the difference and the body mass index. This variability in responses to ambient temperature between the obese and non-obese subjects could be due to a variable influence of heat on arm blood flow consequent on differences in amounts of subcutaneous fat. The ambient temperature for the conduct of the oral glucose tolerance test is important.     

Vasopressin secretion during insulin-induced hypoglycaemia: exaggerated responses in people with type 1 diabetes

Insulin hypoglycaemia causes a rise in plasma vasopressin concentrations in man and the rat, and vasopressin stimulates glucagon secretion and increases hepatic glucose output in man. Vasopressin has also been suggested to have an important synergistic role with corticotrophin releasing factor in the release of adrenocorticotrophin hormone, and a counter-regulatory role for the hormone has been proposed. As diminished anterior pituitary hormone responses to hypoglycaemia have been reported in diabetes mellitus, we studied the plasma vasopressin responses to insulin-induced hypoglycaemia in 10 patients with established Type 1 diabetes and 10 matched control subjects. Blood glucose fell from 4.5 +/- 0.3 to 1.6 +/- 0.1 mmol l-1 (p less than 0.001) in the diabetic group and from 4.6 +/- 0.2 to 1.5 +/- 0.2 mmol l-1 (p less than 0.001) in control subjects, with delayed blood glucose recovery in the diabetic patients. Plasma vasopressin rose in the diabetic patients from 0.9 +/- 0.2 to 6.9 +/- 2.8 pmol l-1 (p less than 0.001), a significantly greater rise (p less than 0.05) than in the control subjects, 0.8 +/- 0.1 to 2.4 +/- 1.0 pmol l-1 (p less than 0.001). Plasma osmolalities remained unchanged and haemodynamic changes were similar in both groups. There is an exaggerated rise in plasma vasopressin concentrations in diabetic patients in response to insulin-induced hypoglycaemia. The putative mechanisms and potential significance of the exaggerated rise are discussed.     

Regulation of non-esterified fatty acid and glycerol concentration by insulin in normal individuals and patients with type 2 diabetes.

Plasma glycerol and non-esterified fatty acid (NEFA) concentrations were determined in the basal state and in response to physiological hyperinsulinaemia in 30 non-obese individuals, 15 with Type 2 diabetes and 15 with normal glucose tolerance. Patients with Type 2 diabetes had higher basal concentrations of both glycerol (81 +/- 7 (+/- SE) vs 61 +/- 7 mumol l-1, p less than 0.05) and NEFA (842 +/- 40 vs 630 +/- 46 mumol l-1, p less than 0.002). Plasma NEFA and glycerol concentrations fell in both groups when steady-state plasma insulin concentrations were raised to approximately 450 pmol l-1 by an infusion of exogenous insulin, but plasma concentrations of glycerol (28 +/- 3 vs 13 +/- 3 mumol l-1, p less than 0.002) and NEFA (186 +/- 15 vs 109 +/- 14 mumol l-1, p less than 0.001) were still higher in patients with Type 2 diabetes. Percentage decrease in glycerol from basal levels in response to insulin was significantly less in patients with Type 2 diabetes than in control subjects (64 +/- 3 vs 80 +/- 3%, p less than 0.005); percentage decrease in plasma NEFA concentration was similar in the two groups (78 +/- 3 vs 80 +/- 4%). These results suggest that both plasma glycerol and NEFA concentrations are higher than normal in patients with Type 2 diabetes when measured at the same insulin concentration, both under basal conditions and in response to physiological hyperinsulinaemia.(ABSTRACT TRUNCATED AT 250 WORDS)     

The outcome of advanced chronic nephropathy in type 1 and type 2 diabetic and non-diabetic patients: a prospective study.

To compare end-stage progression of nephropathy in type 1 and type 2 diabetic patients and non-diabetic subjects, we prospectively studied 92 patients with advanced uraemia not yet on dialysis (mean age 57.2 +/- 15.0 years), with a serum creatinine level above 200 mumol/L. The study included monthly serum creatinine (SC) measurements and quarterly outpatient follow-up (mean 10.8 +/- 7.1 months, range 1-21). Sixty subjects (65.2%) were diabetic (28 type 1 and 32 type 2). At inclusion, 95.6% of patients had anti-hypertensive medications. Drug category, dosage and combination were similar for both groups. Blood pressure (< or = 130/85 mmHg) and glucose level targets (fasting < or = 7.5 mmol/L and postprandial < or = 10 mmol/L) were obtained in all patients. Initial SC was not significantly different between diabetic and non-diabetic patients (426.5 +/- 189.4 mumol/L vs. 405.1 +/- 201.9 mumol/L). SC increased significantly faster in diabetic than non-diabetic patients (respectively 3.9 +/- 6.1% and 1.5 +/- 4.6% monthly, p < 0.05), with no difference between type 1 and type 2 diabetes. One-third (33.7%) of all patients started dialysis during follow-up (40% diabetic and 22% non-diabetic). Their weight, body mass index, age, sex ratio, treatment and aetiology were similar. During follow-up, the patients (29.4%) who sustained a major vascular event differed only in age (62.1 years vs. 55.2 years; p < 0.001). In this study, diabetic renal disease worsened significantly faster than other nephropathies, in spite of proper normalisation of blood pressure and glucose level. Therefore, it is essential to diagnose and manage Type 2 diabetes early to avoid encumbering dialysis centres with older patients.     

Decreased nitric oxide end-products and its relationship with high density lipoprotein and oxidative stress in people with type 2 diabetes without complications

Lipid peroxides are thought to be formed by free radicals and may play an important role in the development of atheromatous vascular diseases. The relationship between serum lipids, lipoproteins, lipid peroxides [thiobarbituric acid reactive substances (TBARS)] and erythrocyte antioxidant enzymes [catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD)] was investigated in non-insulin-dependent diabetic patients with and without coronary heart disease (CHD), and a comparison was made for all the above parameters with non-diabetic patients with CHD. Lipid peroxide concentrations were significantly increased in both groups of diabetic patients and also in non-diabetic patients with CHD, compared to those in control subjects. Diabetic patients with CHD had higher levels of TBARS compared to those diabetics without CHD. Hyperlipidaemia and abnormal lipoprotein levels were observed in all three groups of patients. Increased total cholesterol and LDL-cholesterol were observed in diabetics with CHD compared to those without CHD. Among the erythrocyte antioxidant enzymes, CAT activity was increased, GPx activity was decreased and no change was observed in SOD activity in both groups of diabetic patients and non-diabetic patients with CHD compared to those in controls. A clear correlation was observed between the CAT activity and lipid peroxide concentrations in all the diabetic patients. These observations suggest that there are similar abnormalities in lipid metabolism and erythrocyte antioxidant enzymes in diabetic patients and non-diabetic patients with CHD.     

Marked impairment of the effect of hyperglycaemia on glucose uptake and glucose production in insulin-dependent diabetes

The effect of hyperglycaemia per se on glucose utilization and glucose production was evaluated in 12 patients with insulin-dependent diabetes and in 9 non-diabetic control subjects. In diabetic patients normoglycaemia was maintained during the night preceding the study by a variable intravenous insulin infusion. During the study endogenous insulin secretion was suppressed by somatostatin (300 micrograms h-1) and replaced by infusion of insulin (0.2 mU kg-1 min-1). Glucose utilization and hepatic glucose production rates were quantified at two plasma glucose concentrations (6.7 and 16.7 mmol l-1) using the two-step sequential hyperglycaemic clamp technique in combination with 3-3H-glucose tracer infusion. Duration of each step was 120 min. In diabetic patients glucose utilization, at a glucose concentration of 6.7 mmol l-1, was not different from normal (mean +/- SE: 2.9 +/- 0.2 vs 3.6 +/- 0.3 mg kg-1 min-1, 0.05 less than p less than 0.10), but the response to marked hyperglycaemia was significantly reduced (5.4 +/- 0.5 vs 9.4 +/- 1.0 mg kg-1 min-1, p less than 0.01). Hepatic glucose production was also normal at 6.7 mmol l-1 (1.4 +/- 0.1 vs 1.4 +/- 0.1 mg kg-1 min-1, NS), but whereas in control subjects glucose production was suppressed during hyperglycaemia of 16.7 mmol l-1 (0.3 +/- 0.4 mg kg-1 min-1, p less than 0.01), a slight increase was observed in diabetic patients (2.0 +/- 0.2 mg kg-1 min-1, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Platelet behaviour and haemostatic variables in type 1 (insulin-dependent) diabetic patients with and without albuminuria.

Abnormalities of haemostasis have been implicated in the development of both large and small vessel disease in diabetes. Platelet behaviour and coagulation factors were studied in 28 non-diabetic control subjects and 81 Type 1 diabetic patients with different degrees of albuminuria. Twenty-four (30%) patients had macro- or micro-albuminuria. These patients had elevated levels of beta-thromboglobulin compared with normo-albuminuric patients and control subjects (macro-albuminuric 113 (range 60-314), micro-albuminuric 88 (50-220), normo-albuminuric 55 (13-273), control 52 (18-210) micrograms l-1, p less than 0.001). Similar results were found for platelet factor 4 (macro-albuminuric 57 (9-350), micro-albuminuric 78 (12-205), normo-albuminuric 10 (2-135), control 9 (3-95) micrograms l-1, p less than 0.001). There were decreased beta-thromboglobulin:platelet factor 4 ratios in the albuminuric patients compared with control subjects and normo-albuminuric patients (p less than 0.001). There is abnormal platelet activity in Type 1 diabetic patients with elevated albumin excretion rates.     

Hypoglycaemia unawareness in type 1 diabetes: a lower plasma glucose is required to stimulate sympatho-adrenal activation.

To investigate the relationship between awareness of symptoms and the autonomic reaction of hypoglycaemia, acute hypoglycaemia was induced with intravenous insulin (2.5 mU kg-1 min-1) in diabetic and non-diabetic subjects, all of whom had normal cardiovascular autonomic function tests. Three groups were studied: (1) nine patients with Type 1 diabetes with loss of awareness of hypoglycaemia; (2) eight patients who had normal awareness of hypoglycemia, matched for duration of diabetes and blood glucose control; (3) eleven non-diabetic volunteers. The onset of the acute autonomic reaction was identified objectively by the sudden and rapid responses of heart rate and sweating. Cognitive function and hypoglycaemia symptom scores were estimated serially. Acute autonomic activation was observed to occur in all subjects in response to hypoglycaemia. In the 'unaware' diabetic patients, onset of the reaction occurred at a significantly lower plasma glucose (1.0 +/- 0.1 mmol l-1) than in the 'aware' diabetic patients (1.6 +/- 0.2 mmol l-1) (p less than 0.05) or in the non-diabetic control group (1.4 +/- 0.1 mmol l-1) (p less than 0.05). Obvious neuroglycopenia was observed only in the 'unaware' diabetic group and developed when plasma glucose had declined to approximately 1.4 +/- 0.1 mmol l-1, and thus preceded the reaction (p less than 0.02 vs the autonomic threshold). The maximal rise in plasma adrenaline was of similar magnitude in all three groups but a lower plasma glucose was required to stimulate this hormonal response in the 'unaware' patients, in whom the plasma adrenaline concentration was lower at the time of the reaction. Thus, the plasma glucose at which activation of the autonomic reaction was observed was lower in the diabetic patients with unawareness of hypoglycaemia.     

Contrasting fibrinolytic responses in type 1 (insulin-dependent) and type 2 (non-insulin-dependent) diabetes.

To study fibrinolysis in relation to microvascular diabetic complications, 20 control subjects were compared with 50 Type 1 (insulin-dependent) diabetic patients of similar age, 20 with no complications, 17 with laser-treated retinopathy, and 13 with neuropathy and retinopathy. None were smokers, hypertensive or had macrovascular disease. Pre- and post-venous occlusion blood samples for tests of fibrinolysis were taken. Median (interquartile range) basal tissue plasminogen activator (t-PA) activity was lower in control subjects (100 (less than 100-100) IU l-1) than diabetic patients (uncomplicated 145 (100-280) IU l-1, p = 0.015; retinopathy 180 (100-228) IU l-1, p = 0.037; neuropathy 210 (125-310) IU l-1, p = 0.004, respectively). Basal t-PA inhibition (PAl-1 activity) was higher in control subjects (5.9 (4.5-9.5) kIU l-1) than diabetic patients (uncomplicated 4.0 (3.3-5.0) kIU l-1, p = 0.001; retinopathy 4.5 (3.1-6.3) kIU l-1, p = 0.058; neuropathy 4.0 (3.0-5.4) kIU l-1, p = 0.015, respectively). Post-venous occlusion t-PA antigen was higher in control subjects (10.2 (7.3-15.1) micrograms l-1) than neuropathic patients (5.5 (4.9-7.3) micrograms l-1, p = 0.004). Other tests showed a consistent, but non-significant, trend towards increased basal fibrinolysis in the Type 1 diabetic patients. The results indicate that Type 1 diabetic patients have enhanced basal fibrinolysis. The diminished response to venous occlusion in neuropathic patients is consistent with an endothelial cell defect.(ABSTRACT TRUNCATED AT 250 WORDS)     

Carbohydrate and lipid metabolism of skeletal muscle in type 2 diabetic patients

Peripheral hyperinsulinaemia is the cause of metabolic changes that might contribute to the high incidence of macrovascular disease in patients with diabetes mellitus. In order to test this hypothesis muscle biopsies from 12 Type 2 diabetic patients and 14 age and sex matched non-diabetic patients, undergoing minor surgery, were obtained. The diabetic patients had significantly elevated fasting serum insulin (0.29 +/- 0.05 vs 0.06 +/- 0.03 nmol-1) and glucose (8.3 +/- 1.5 vs 4.6 +/- 0.5 mmol-1) and HbA1 levels (8.4 +/- 0.4 vs 5.0 +/- 0.2 per cent). The fasting and 2-h postprandial C-peptide levels were 0.99 +/- 0.25 vs 0.39 +/- 0.12 and 3.12 +/- 0.75 vs 1.09 +/- 0.34 nmol/l, respectively. The diabetic patients showed a marked elevation of triglyceride in the striated muscle biopsies compared to the non-diabetic controls (290 +/- 52 vs 48 +/- 6 mumol/g wet weight, p less than 0.001). Moreover, the activities of glucose-6-phosphate dehydrogenase (0.25 +/- 0.03 vs 0.13 +/- 0.01 U/g wet weight) and malic enzyme (0.15 +/- 0.01 vs 0.05 +/- 0.01 U/g wet weight), necessary for lipid synthesis, were significantly increased (both p less than 0.001) in the diabetic patients while the glycolytic enzymes, hexokinase (0.65 +/- 0.09 vs 1.82 +/- 0.11 U/g wet weight), pyruvate kinase (7.3 +/- 0.9 vs 13.2 +/- 0.9 U/g wet weight), phosphofructokinase (1.3 +/- 0.2 vs 2.6 +/- 0.2 U/g wet weight), and alpha-glycerophosphate dehydrogenase (7.3 +/- 0.5 vs 12.5 +/- 0.7 U/g wet weight) were decreased (all p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

A Comparison of Different Methods of Assessing Free Radical Activity in Type 2 Diabetes and Peripheral Vascular Disease

Increased free radical activity in diabetes mellitus may contribute to the higher prevalence and mortality from macrovascular disease in diabetic patients. To investigate this, levels of plasma antioxidants (superoxide dismutase, caeruloplasmin, plasma, and lysate thiol), diene conjugates, lipid peroxides, and chemiluminescence were measured in diabetic and non-diabetic patients with peripheral vascular disease compared with healthy control subjects. Caeruloplasmin, diene conjugate ratio, and lipid peroxides were significantly increased in patients with vascular disease but there was no difference between diabetic and non-diabetic patients. Conjugated diene ratio correlated with caeruloplasmin (r = 0.40, p < 0.02) and inversely with superoxide dismutase level (r = ?0.36, p < 0.05) but there was no significant correlation between other antioxidants and diene conjugates, lipid peroxides or chemiluminescence. The relationship between different indirect measurements of free radical activity is variable but there appears to be no additive effect of diabetes on the increased free radical activity associated with vascular disease.     

Neuropsychological performance differs between type 1 diabetic and normal men during insulin-induced hypoglycaemia.

Cerebral function was measured with a neuropsychological test battery before, during, and after insulin-induced hypoglycaemia (blood glucose approximately 2.0 mmol l-1) in 10 male Type 1 diabetic patients (age 20-43 years, duration of diabetes 14 (2-30) years) and in 12 normal men. There were no group differences in neuropsychological results at normal glucose levels. Significant effects of hypoglycaemia were found in reaction-time measures (p less than 0.001) and in other tests requiring speed and attention (p less than 0.001), in verbal fluency (p less than 0.05), and short-term memory (p less than 0.001). Significant group effects and interactions (p less than 0.05) revealed that the diabetic patients were generally more affected by hypoglycaemia than the normal subjects. This might have been partly due to the larger absolute decrease in blood glucose level in the diabetic patients, although the rate of glucose decrease was not related to performance in either group. Thus, the diabetic brain might be more vulnerable to hypoglycaemia, perhaps through the persistent impact of repeated hypoglycaemic episodes, although no neuropsychological deficit is demonstrable at normal blood glucose levels.     

Clinical macrovascular disease in Caucasoid diabetic subjects: logistic regression analysis of risk variables

Summary A cross-sectional study of 1084 Caucasoid diabetic subjects in rural Western Australia revealed a high rate of clinical macrovascular disease (46%), including coronary heart disease (13%), stroke (8%), and peripheral vascular disease (38%). Age was the major time-related variable for total macrovascular disease and for peripheral vascular disease, with identical prevalence rates in Type 1 (insulin-dependent) and Type 2 (non-insulin-dependent) diabetes when age was taken into account. In 179 Type 1 diabetic subjects, logistic regression analysis showed no associated risk factors other than age. In 905 Type 2 diabetic subjects the independent risk factors for total macrovascular disease, identified by a forward stepwise selection procedure, were age as the major contributor, with plasma creatinine levels and plasma glucose levels (all p<0.001), high-density lipoprotein cholesterol levels, serum total cholesterol levels, and the (supine-erect) systolic blood pressure difference (all p<0.05). There were no direct associations with percentage desirable weight, cigarette smoking or male sex. Type 2 diabetic subjects demonstrated a very strong negative association between high-density lipoprotein cholesterol levels and coronary heart disease, and significant associations were found also between plasma glucose levels and coronary heart disease (p<0.01), and glycosylated haemoglobin levels and peripheral vascular disease (p<0.001).     

The effect of body weight on insulin activity

Insulin resistance is found patients with diabetes mellitus type 2 as well as in obese subject without diabetes. The objective of our investigation was to compare the action of insulin in morbidly obese subject with and without diabetes and in diabetic subject with different degrees of obesity. A total of 36 diabetic were examined, divided according to the BMI into morbidly obese (DMTO: BMI > 40 kg/m-2.n = 6) those with medium severe obesity (DMSO: BMI 31-40 kg.m-2.n = 16), with slight overweight DMLO. BMI 26-91 kg.m-2.n = 9) and non-obese diabetics (DMBO). BMI 21-26 kg.m-2.n = 5). The group of morbidly obese non-diabetic subject (NDTO, BMI > 40 kg.m-2.n = 5) and non-obese healthy subject (C, BMI < 26 kg.m-2, n = 12) served as control. All examined subject were of similar age the diabetic subject had similar values of indicator of diabetic control (HbA1c was 7.1 +/- 0.5%). The examination was made using the method of an isoglycaemic hyperinsulinaemic clamp on a Biostator at an insulin infusion rate of 1mU.kg-1.min-1 for a period of 20 minutes. The results of the index of tissue sensitivity to insulin revealed a markedly deteriorated action of insulin in morbidly obese diabetes and non-diabetics in relation to control group of healthy slim controls (M/I, DMTO: 12.4 +/- 7.3 and NDTO: 9.2 +/- 4.1, p < 0.001, mumol.kg-1.min-1 na mU.l-1 x 100), in midly and medium obese diabetics the insulin resistance was of difference grades (M/I, DMLO: 34.2 +/- 9.3, p < 0.05, and DMSO: 25.9 +/- 18.5 p < 0.001 mumol.kg-1.min-1 na mU.l-1 x 100. Non-obese diabetic and non-diabetic subject had a normal insulin action (M/I, DMBO: 58.3 +/- 29.4 and C: 48.9 +/- 5.0 mumol.kg-1.min-1 per mU.l-1 x 100. The metabolic glucose clearance differed however between diabetic and non-diabetic subject (MCRG, DMTO: 2.0 +/- 0.4, p < 0.001, DMSO: 3.8 +/- 2.4, p < 0.001, DMSO: 5.4 +/- 1.7, p < 0.05 v.s. C: 8.6 +/- 1.1 and NDTO: 3.8 +/- 1.5, p < 0.001 ml.kg-1.min-1). The statistical significance is related to the control group of slim healthy subject. From this ensues that no significant difference was found between slim diabetic and non-diabetic subjects in the majority of parameters expressing the action of insulin with the exception of the metabolic glucose clearance. At the same time the authors found in the whole group of 53 examined subject a statistically significant correlation between the BMI and the index of tissue sensitivity for insulin (M/I) (r = -0.55, p < 0.001). On examination of characteristics of insulin receptors on erythrocytes the authors found a reduced number in diabetic subject as compared with the two control groups (p < 0.05). It may thus be concluded from this investigation that the BMI has a decisive role in the action insulin.     

Leucocyte mobilization and release of neutrophil elastase following acute insulin-induced hypoglycaemia in normal humans

Insulin-induced hypoglycaemia in humans is associated with the rapid mobilization of leucocytes in peripheral blood. The aim of the present study was to determine whether neutrophil activation, manifested in plasma by neutrophil elastase concentration, occurs in response to insulin-induced hypoglycaemia. Acute hypoglycaemia (mean blood glucose 1.3 +/- 0.2 mmol l-1; mean +/- SD) was induced with intravenous insulin in 15 normal human subjects, and provoked an increase in the neutrophil count from 3.4 (range 1.9-6.5) to 10.7 (9.4-16.3) X 10(9) l-1 (p less than 0.001), and in the total leucocyte counts from 5.7 (4.1-8.1) to 12.8 (11.3-18.6) X 10(9) l-1 (p less than 0.001), with associated elevations in plasma neutrophil elastase concentration from 21 (12-34) to 29 (14-70) micrograms l-1 (p less than 0.05), and in total neutrophil elastase concentration from 5.90 (3.13-8.20) to 25.20 (23.00-52.00) mg l-1 (p less than 0.001). As neutrophil elastase is implicated in the development of vascular disease, this rise in response to hypoglycaemia may be of pathological importance in insulin-treated diabetic patients.     

Effects of intensive dietary treatment on insulin-stimulated skeletal muscle glycogen synthase activation and insulin secretion in newly presenting type 2 diabetic patients

Ten newly presenting, untreated, Europid Type 2 diabetic patients were studied before and after 8 weeks treatment with intensive diet alone. Nine normal control subjects were also studied. The degree of activation of skeletal muscle glycogen synthase (GS) was used as an intracellular marker of insulin action, prior to and during a 240-min insulin infusion (100 mU kg-1 h-1). Fasting blood glucose decreased from 12.1 +/- 0.9 (+/- SE) to 9.2 +/- 0.8 mmol l-1 (p less than 0.01), but there was no change in fasting insulin concentrations, 9.9 +/- 2.3 vs 9.3 +/- 2.1 mU l-1. Fractional GS activity did not increase in the Type 2 diabetic patients during the insulin infusion either at presentation (change -1.5 +/- 1.9%) or after treatment (change +0.9 +/- 1.8%), and was markedly decreased compared with the control subjects (change +14.5 +/- 2.8%, both p less than 0.001). Glucose requirement during the clamp was decreased in the Type 2 diabetic patients at presentation (2.2 +/- 0.7 vs 7.3 +/- 0.6 mg kg-1 min-1, p less than 0.001), and despite improvement following dietary treatment to 3.3 +/- 0.6 mg kg-1 min-1 (p less than 0.01) remained lower than in the control subjects (p less than 0.001). Fasting plasma non-esterified fatty acid (NEFA) concentrations were elevated at presentation (p less than 0.05), and failed to suppress normally during the insulin infusion. After treatment fasting NEFA concentrations decreased (p less than 0.05) and suppressed normally (p less than 0.05). Insulin secretion was assessed following an intravenous bolus of glucose (0.5 g kg-1) at euglycaemia before and after treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Self-collection by diabetic patients of capillary blood for free insulin monitoring; reduction by diamide of haemolysis-induced insulin loss.

The need to precipitate bound insulin immediately after withdrawal of blood and the tendency to haemolysis, which reduces immunoassayable insulin, have prevented development of methods of self-collection of capillary blood for later free insulin measurement. We therefore investigated the use of the thiol-oxidizing agent, diamide, to prevent insulin loss with haemolysis and developed a self-collection procedure with capillary tubes pre-filled with diamide and polyethyleneglycol (PEG, for separation of free and bound insulin). Diamide (final concentration 5 mmol l-1) reduced serum insulin loss from 48 +/- 4 (+/- SE) to 11 +/- 4% (p less than 0.001) in maximally-haemolysed samples. The effect of diamide was concentration-dependent up to 5 mmol l-1. Diamide had no effect on the standard curve for radioimmunoassay of insulin. Levels of serum free insulin in self-collected capillary blood were significantly correlated with venous serum free insulin in 22 non-diabetic subjects (r = 0.92, p less than 0.001), 52 Type 1 diabetic patients (r = 0.86, p less than 0.001), and 18 Type 2 diabetic patients (r = 0.97, p less than 0.001). Mean capillary free insulin concentration was higher than in venous serum (22% in normal subjects, 64% in Type 1, and 23% in Type 2 diabetic patients). Storage at room temperature of capillary blood containing PEG/diamide for 72 h did not alter immunoassayable insulin concentrations.