Acute actions of sulfonylurea drugs during long-term treatment of NIDDM.

The acute effect of sulfonylurea drugs during long-term treatment was evaluated in two separate studies. In the first study, the levels of plasma glucose, insulin, and C-peptide were measured after intake of 10 mg glyburide alone or together with a standardized mixed meal in 10 non-insulin-dependent diabetes mellitus (NIDDM) patients treated with 10-20 mg glyburide/day for greater than 2 yr. There was no acute effect of glyburide on the insulin and C-peptide responses to the meal or during continued fasting, indicating the absence of an acute insulinotropic action of glyburide during chronic treatment. The glucose increase after the meal was also unchanged, but a significant glucose reduction was found after glyburide intake during continued fasting, suggesting a sustained acute extrapancreatic (hepatic) effect. In the second study, the diurnal glycemic excursions in 8 NIDDM patients chronically and continuously (24 h/day) exposed to glipizide (2.5-7.5 mg 3 times/day) were similar when the drug was taken 30 min before each of the three main meals and when taken immediately before the meals. It is concluded that there is no acute insulinotropic action of sulfonylurea drugs during chronic continuous exposure, whereas an acute extrapancreatic action may prevail. During such exposure, there seems to be no clinical benefit in taking a sulfonylurea 30 min before rather than at the start of a meal.

Decrease in beta-receptor density explaining the development of pindolol- and prenalterol-tolerance in orthostatic hypotension.

A 58-year-old woman with diabetic autonomic dysfunction was well treated for orthostatic hypotension with pindolol. After eight months however, symptoms recurred in spite of continued treatment with pindolol. Addition of prenalterol, a beta-blocking agent with very high intrinsic sympathomimetic activity, had a marked clinical effect on her orthostatic hypotension. Before treatment with prenalterol, stroke volume and left ventricular enddiastolic volume markedly decreased during tilt, as demonstrated by radionuclide angiography. After short-term prenalterol treatment, the orthostatic decreases of stroke volume and left ventricular end-diastolic volume were less pronounced. During continued prenalterol treatment, both symptoms and haemodynamic changes recurred in the erect position. Beta-receptor density in her lymphocytes decreased from elevated levels before prenalterol to subnormal levels, when the clinical effect had disappeared. The data suggest that the tolerance development to prenalterol may be explained by the decrease in beta-adrenergic receptor density.

Insulin binding and action on fat cells from young healthy females and males.

Insulin binding and action were studied in fat cells from the gluteal region of young healthy subjects. Fat cells from females were larger than those of males, had higher insulin receptor binding and higher rates of noninsulin-stimulated and maximally insulin-stimulated rates of methylglucose transport and glucose metabolism when these data were expressed per cell number. However, when insulin binding and insulin effects were expressed per cell surface, which may be physiologically more relevant, no sex differences were found in insulin binding and glucose transport, whereas noninsulin-stimulated and maximally insulin-stimulated glucose metabolism was still significantly increased in female fat cells. The latter indicates postreceptor differences in glucose metabolism between females and males. The insulin concentrations causing half-maximal responses (a measure of the sensitivity to insulin) of glucose transport, glucose metabolism and lipolysis were similar in fat cells from the two sexes, which is consistent with the comparable values of insulin receptor binding when adjusted to cell surface. Studies of rate-determining steps for the glucose utilization of human fat cells showed that glucose transport was not the rate-limiting step at physiological glucose concentrations. Moreover, at physiological glucose levels, glucose metabolism exhibited a decreased maximal insulin responsiveness and an increased insulin sensitivity when compared with glucose metabolism at low glucose concentrations at which glucose transport is rate limiting for the fat cell glucose utilization.

Circadian profiles of insulin receptors in insulin-dependent diabetics in usual and poor metabolic control.

We have characterized the 24-h changes of insulin receptors on erythrocytes from patients with insulin-dependent diabetes of long duration. These diabetics were studied both in usual and poor metabolic control. Moreover, we have examined daytime changes of insulin receptors on monocytes from newly discovered diabetics. In both erythrocyte and monocyte studies, diabetics were compared to healthy controls. At insulin tracer concentration, insulin receptor binding to erythrocytes from diabetics in usual control and normal volunteers underwent a statistically significant diurnal variation with high binding values in the early morning, low daytime values with a nadir in the late afternoon, and a peak around midnight. Even diabetics in poor metabolic control due to insulin deprivation had preserved a similar 24-h rhythm of erythrocyte insulin receptors. Insulin receptor binding to monocytes at insulin tracer concentration declined significantly during the day both in newly discovered diabetics and in healthy controls. The mechanisms responsible for the acute phase changes of insulin-receptor binding are unknown, but the receptor changes seem related to the fed state. Moreover, analysis of the temporal interrelationship between erythrocyte insulin binding and plasma insulin concentration in diabetics during the 24-h period suggests that in these patients insulin may be one of the factors determining the rapid insulin receptor regulation.

Insulin receptor binding and receptor-mediated insulin degradation in human adipocytes.

UNLABELLED: 125I-insulin binding and receptor-mediated insulin degradation were studied in isolated human fat cells from subcutaneous tissue. A high albumin concentration during cell isolation and incubation protected the fragile human adipocyte from lysis. Binding of tracer was pH dependent with an optimum between 7.4 and 7.6. At 37 degrees C steady state was reached by 45 min and maintained for at least 2 h. The binding of labelled insulin in the presence of 10 mumol/l unlabelled insulin was only 1-4% of the total insulin binding. The half-maximal displacement of tracer iodoinsulin (10 pmol/l) by unlabelled insulin occurred at 0.25 nmol/l. Kinetic studies of the dissociation of labelled iodoinsulin from fat cells showed a slight acceleration in the presence of a high concentration of unlabelled insulin in the washout buffer as compared to a buffer containing no insulin. At steady state binding about 95% of the cell-associated radioactivity was extracted as iodoinsulin as judged by gel filtration. The remaining 5% co-eluted with iodotyrosine. During 60 min about 90% of the cell-associated radioactivity dissociated as iodoinsulin and the rest as iodotyrosine. CONCLUSIONS: 1) A high albumin content of buffers prevents traumatization of the human adipocyte; 2) under these conditions steady state binding of insulin is readily measured at 37 degrees C; 3) the use of a washing procedure makes the non-specific binding negligible; 4) the human adipocyte insulin receptor has a very high affinity; 5) receptor-mediated insulin degradation is minimal.

Impaired cellular insulin binding and insulin sensitivity induced by high-fructose feeding in normal subjects.

We have studied whether the sucrose-induced reduction of insulin sensitivity and cellular insulin binding in normal man is related to the fructose or the glucose moiety. Seven young healthy subjects were fed their usual diets plus 1000 kcal extra glucose per day and eight young healthy subjects were fed their usual diets with addition of 1000 kcal extra fructose per day. The dietary regimens continued for 1 week. Before change of diet there were no statistically significant differences between body weight and fasting plasma concentrations of glucose, insulin, and ketone bodies in the two groups studied. High-glucose feeding caused no significant changes in insulin binding or insulin sensitivity whereas high-fructose feeding was accompanied by a significant reduction both of insulin binding (P less than 0.05) and insulin sensitivity (P less than 0.05). The changes in insulin binding and insulin sensitivity correlated linearly (r = 0.52, P less than 0.01). We conclude that fructose seems to be responsible for the impaired insulin binding and insulin sensitivity induced by sucrose.

Diagnosis of Mycoplasma hominis pyelonephritis by demonstration of antibodies in urine.

To evaluate the diagnostic significance of the demonstration in urine of antibodies to Mycoplasma hominis, 1,000 samples of urine with more than 5 leukocytes per high-power field were serologically investigated by indirect hemagglutination, using glutaraldehyde-fixed erythrocytes coated with M. hominis antigen. The samples were collected from 702 patients. Antibodies were demonstrated in the urine of nine patients, all of whom had signs of acute attack of pyelonephritis. In seven of these patients, characterized by mild or moderate clinical signs and absence of lower urinary tract symptoms, bacterial causes were not observed, whereas M. hominis organisms were isolated from the upper urinary tract in most cases and from the bladder urine in all cases. In two patients, characterized by severe clinical signs and presence of lower urinary tract symptoms, both M. hominis and bacteria were isolated from the upper urinary tract and ballder urine. The demonstration of antibodies to M. hominis in urine is of high diagnostic value as they were only observed in patients in whom M. hominis infection in the upper urinary tract was evident or likely and only in the presence of clinical signs of acute attacks of pyelonephritis.

Increased insulin sensitivity and cellular insulin binding in obese diabetics following treatment with glibenclamide.

The aim of the present study was to examine the effect of glibenclamide on the insulin receptors, the insulin sensitivity and the insulin secretion in obese non-ketotic diabetics. Two groups of 9 obese diabetics were studied before and after 10 days' treatment with a 1200 kcal's diet and a 1200 kcal's diet + 10 mg/day of glibenclamide, respectively. In the group treated with diet alone we found no significant alteration of the insulin secretion pattern (P greater than 0.1). However, the insulin sensitivity increased 37% (P less than 0.01). Furthermore, the insulin binding to monocytes increased (P less than 0.01) due to a 36% rise of the binding affinity. In the group treated with glibenclamide and diet the insulin secretory pattern was unchanged, too (p greater than 0.1). The insulin sensitivity, however, increased 83% (P less than 0.01). Moreover, the insulin binding was raised (P less than 0.01) as a result of a 80% rise of the number of insulin receptors. In 4 patients who were treated with diet (1200 kcal/day) plus glibenclamide and in 5 patients who were treated with diet alone (1200 kcal/day) the insulin binding to monocytes was studied during treatment for 1 year. After 1 year we found a significantly (P less than 0.005) higher cellular insulin binding in the glibenclamide treated patients compared to the patients who got diet alone. We conclude that 1) the augmentation of the insulin sensitivity is of great importance for the normalization of the diabetic state in obese, 2) the increase in insulin binding may be of importance for the increase in insulin sensitivity, 3) glibenclamide appears to enhance the insulin sensitivity through an increase in the number of insulin receptors.

Normalization of the insulin sensitivity and the cellular insulin binding during treatment of obese diabetics for one year.

The relative importance of the insulin resistance, the decreased cellular insulin binding and the relative insulin deficiency in the pathogenesis of diabetes mellitus in obese subjects has been studied. Ten obese diabetics were studied before and during treatment for 1 year with a 1200-1500 kcal's diet. No change was found in the insulin response to iv injection of glucose during treatment (P greater than 0.1), whereas the insulin sensitivity was normalized after 1 year (P less than 0.01). In parallel to the clinical normalization and the improvement of the insulin sensitivity the insulin binding to monocytes was normalized (P less than 0.01). We conclude that both the insulin resistance and the relative insulin deficiency are of decisive importance in the pathogenesis of diabetes mellitus of the obese. The insulin receptor defect seems to be one of the major factors responsible for the insulin resistance.