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Helmuth Haslacher,1 Hannelore Fallmann,2 Claudia Waldhäusl,3 Edith Hartmann,2 Oswald F Wagner,1 Werner K Waldhäusl2,41Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria; 2Rehabilitation Clinic for Diabetes and Metabolic Diseases, Moorbad Neydhartig, Neydharting, Upper Austria, Austria; 3Department of Radiotherapy, Medical University of Vienna, Vienna, Austria; 4Department of Medicine III, Medical University of Vienna, Vienna, AustriaPurpose: To explore differences in baseline characteristics following three weeks of semi-standardized in-patient care between patients with obesity without and with type 2 diabetes (T2D).Patients and methods: Patients without or with T2D were matched according to age, sex, and BMI. Food intake was restricted to 1,200–1,600 kcal/d to which a 400–600 kcal/d exercise load was added, and data were compared using Student’s t-test, general linear models, and Spearman-rank correlations.Results: At baseline, patients with obesity and T2D displayed, besides elevated blood glucose and HbA1c values, higher serum liver enzymes (p

Objective: Plasma free fatty acid (FFA) concentrations are increased in states of insulin resistance. Therefore, this study evaluated apoptosis and underlying mechanisms induced by selected nutritional FFAs, a defined FFA-mix, and human plasma containing high FFA concentrations in human smooth muscle cells (HSMCs)., Research Design and Methods: HSMCs were incubated (24-72 h) with selected FFAs (100-300 micromol/l), an FFA-mix (palmitic-/stearic-/oleic-/linoleic-/alpha-linolenic acid=2.6/1/3.6/9/1; 300-900 micromol/l), or with high FFA-plasma (600 micromol/l) versus respective control cultures. Apoptosis, caspase activation, and protein expression were determined by DNA-fragmentation assays, flow cytometry, and Western blots, respectively., Results: Exposure (24h) of HSMCs to 300 micromol/l stearic-, oleic-, linoleic-, alpha-linolenic-, and arachidonic acid induced apoptosis, correlating (p<0.01) with the FFAs' chain length (r=0.602) and number of FFA double bonds (r=0.956). After 48 h, 100 micromol/l of all tested FFAs - including palmitic acid - were already sufficient to trigger HSMCs' cell death. FFA-exposure resulted in activation of caspases and apoptosis was completely abolished by co-incubation with caspase inhibitors and negatively correlated (p<0.01) with the base-excision repair protein XRCC1 (r=-0.765) and with c-myc's antagonist mad (r=-0.916), whereas positive correlations (p<0.01) were found for protein expression of the proto-oncogene c-myc (r=0.972) and the transcription factor E2F-1 (r=0.971). Exposure of HSMCs to the defined FFA-mix and to plasma samples from individuals with elevated plasma FFAs supported the results obtained by defined FFA stimulation., Conclusions: Since smooth muscle cells surround the macrophage/foam cell/lipid-laden artheromatous core of atherosclerotic lesions with a protective fibrous cap, their FFA-induced HSMC apoptosis could contribute to progression of atherosclerosis by thinning of the fibrous cap and subsequent plaque destabilization.

Lipoic acid was recently demonstrated to improve endothelial dysfunction or retinopathy not only in rats but also in diabetic patients. We tested the hypothesis that R-(+)-alpha-lipoic acid (LA) directly affects human endothelial cell (EC) function (e.g., apoptosis, proliferation, and protein expression), independent of the cells' vascular origin. Macrovascular EC (macEC), isolated from umbilical (HUVEC) and adult saphenous veins and from aortae, as well as microvascular EC (micEC) from retinae, skin, and uterus, were exposed to LA (1 mumol/l-1 mmol/l) with/without different stimuli (high glucose, TNF-alpha, VEGF, wortmannin, LY-294002). Apoptosis, proliferation, cell cycle distribution, and protein expression were determined by DNA fragmentation assays, [(3)H]thymidine incorporation, FACS, and Western blot analyses, respectively. In macro- and microvascular EC, LA (1 mmol/l) reduced (P < 0.05) basal (macEC, -36 +/- 4%; micEC, -46 +/- 6%) and stimulus-induced (TNF-alpha: macEC, -75 +/- 11%; micEC, -68 +/- 13%) apoptosis. In HUVEC, inhibition of apoptosis by LA (500 mumol/l) was paralleled by reduction of NF-kappaB. LA's antiapoptotic activity was reduced by PI 3-kinase inhibitors (wortmannin, LY-294002), being in line with LA-induced Akt phosphorylation (Ser(437), +159 +/- 43%; Thr(308), +98 +/- 25%; P < 0.01). LA (500 mumol/l) inhibited (P < 0.001) proliferation of macEC (-29 +/- 3%) and micEC (-29 +/- 3%) by arresting the cells at the G(1)/S transition due to an increased ratio of cyclin E/p27(Kip) (4.2-fold), upregulation of p21(WAF-1/Cip1) (+104 +/- 21%), and reduction of cyclin A (-32 +/- 11%), of hyperphosphorylated retinoblastoma protein (macEC: -51 +/- 7%; micEC: -50 +/- 15%), and of E2F-1 (macEC: -48 +/- 3%; micEC: -31 +/- 10%). LA's ability to inhibit apoptosis and proliferation of ECs could beneficially affect endothelial dysfunction, which precedes manifestation of late diabetic vascular complications.

This study evaluated direct effects of peroxisome proliferatoractivated receptor gamma(PPARgamma) agonists, including thiazolidinediones (TZDs), on vascular cell apoptosis and related protein expression to test the hypothesis that these effects are dependent on i) the respective agent's structure and ii) endothelial cells' vascular origin. Exposure (48 h) of human umbilical vein endothelial cells (HUVECs, n=6) to up to 10 microM troglitazone (TRO), rosiglitazone, pioglitazone, and to up to 50 microM RWJ241947=MCC-555 (RWJ) inhibited (p<0.05) apoptosis by 8-25%, whereas 15-deoxy-Delta(12-14)-prostaglandin J(2) (PGJ(2)) triggered (50 microM: + 400%, p<0.05) endothelial cell death versus control (=100%). Moreover, RWJ (50 microM) completely abrogated TNF-alpha(2000 U/ml) and stearic acid (200 microM) induced apoptosis in HUVECs . Similar results were obtained in human adult (saphenous) vein- and aortic endothelial cells, the latter showing no anti-apoptotic response to TRO. In HUVECs, TZDs' anti-apoptotic effects inversely correlated (r=-0.95, p<0.01) with increased (p<0.05) expression of the apoptosis-inhibitor bcl-2, whereas PGJ(2)-induced apoptosis was associated with upregulation of c-myc (+447%) and E2F-1 (+339%). Additionally, TZDs (by 25-39%) and PGJ(2) (-70%) reduced (p<0.05) expression of heat shock protein 60 (hsp60) showing no correlation with apoptosis (r=0.14, n.s.). Modulation of apoptosis by PPARgammaagonists differs in endothelial cells dependent on their vascular origin and the agonists' structure. Thiazolidinediones' ability to reduce both, endothelial apoptosis and hsp60 expression could well add to beneficial vascular effects attributed to these oral antidiabetic drugs.

The vascular endothelium, building the inner layer of capillaries and blood vessels of all sorts, represents a highly active metabolic and endocrine organ producing a multitude of different molecules, including vasoactive peptide hormones, growth factors, coagulation factors and adhesion molecules. In addition, it expresses many of the respective endocrine, paracrine and cytokine/growth factor receptors. It thereby regulates the delicate balance between vasoconstriction and vasodilation, between coagulation and fibrinolysis, proliferation and apoptosis, as well as between transient adhesion and diapedesis of blood borne leukocytes. This minireview addresses the potential interactions of these important functions in the states of diabetes and of insulin resistance including obesity, hypertension and dyslipidaemia, all of which characterized by endothelial dysfunction.

To establish a qualitative and quantitative model of blood glucose response to stress hormone exposure, healthy subjects (HS) on and off somatostatin (250 micrograms/h) as well as insulin dependent diabetic patients were infused with either epinephrine (E), glucagon (G), cortisol (F), growth hormone (GH) or with a cocktail of these hormones raising plasma stress hormones to values seen in severe diabetic ketoacidosis. The developed input/output model consists of two submodels interconnected in series plus two additional submodels for correction of gains describing both sensitivity of tissue response and utilisation as well as provision of glucose. It was shown and confirmed experimentally that blood glucose response to stress hormones was essentially nonlinear. Furthermore, the mathematical models for healthy subjects and for insulin dependent diabetic patients proved to be of the same structure and differed only in the values of some typical parameters. The model raises the possibility to describe and in part to predict blood glucose response to stress hormone exposure in healthy man and insulin dependent diabetic patients.

1. In a single-blind, placebo controlled study the influence of tenoxicam on responses of glucose, insulin and C-peptide to oral doses of glucose and glibenclamide was examined in 16 healthy male volunteers. 2. The subjects received once daily doses of 2.5 mg glibenclamide for 12 days. From day 5 through 12 eight subjects received concomitantly 20 mg tenoxicam once daily and the remaining eight subjects received placebo. 3. On days 1, 4, 5 and 12 glibenclamide was taken with 75 g glucose and blood glucose, serum insulin and C-peptide were measured over 5 h. Plasma levels of glibenclamide and tenoxicam (where appropriate) were followed over 10 h. 4. Characteristic parameters of blood glucose and insulin and C-peptide responses did not change significantly with time (day) and there was no difference between both treatment groups. 5. Baseline insulin increased from 11.7 mu l-1 on day 1 to 15.6 mu l-1 on day 4 (P = 0.009), likewise baseline C-peptide increased from 478 pmol l-1 to 530 pmol l-1 (P = 0.05), but there was no further change in the subsequent treatment period. 6. The AUC of the glibenclamide plasma concentration-time curve did not show changes with time or differences between treatment groups. The mean (s.d.) oral clearance of tenoxicam was 2.5 (1.5) ml min-1 and appeared slightly higher than in previous studies. 7. It was concluded that tenoxicam did not affect overall glycoregulation in healthy subjects under glibenclamide steady state conditions.

To determine quantitative and qualitative differences in insulin secretion equimolar amounts of glucose and arginine were infused in 9 healthy subjects, in 8 individuals each with obesity without and with impaired glucose tolerance, and in non-obese and obese non-insulin-dependent diabetic patients (NIDDM). Insulin secretion was calculated after individual determination of metabolic clearance rate of C-peptide (MCRcp) both as the area under the C-peptide concentration curve times MCRcp, and by a mono-compartment mathematical model, both yielding identical results. MCRcp fell consistently with increasing C-peptide infusion rate (e.g.: healthy subjects: C-peptide, 10 nmol/h, 4.2 +/- 0.4; 20 nmol/h, 3.3 +/- 0.3; 30 nmol/h, 3.1 +/- 0.2 ml/kg.min; p less than 0.05 to p less than 0.01). Basal insulin secretion was 2.1-fold greater in the obese with impaired glucose tolerance than in healthy subjects, but was unchanged in non-obese NIDDM. Glucose and arginine triggered insulin release was greater than in healthy subjects at almost identical area under the respective substrate concentration curve (AUC/kg body weight) in obese subjects without (2-fold) and with impaired glucose tolerance (4-fold), and in NIDDMs following i.v. arginine (2-fold). The mean ratio of incremental insulin release to i.v. glucose and arginine was smaller in NIDDM (normal weight, 1.3 +/- 0.4; obese, 1.0 +/- 0.2) than in healthy (2.0 +/- 0.3), or obese subjects with impaired glucose tolerance (2.8 +/- 0.7). Stimulated C-peptide/insulin ratio was reduced in all patients vs that in healthy subjects (p less than 0.05). We conclude that (a) MCR of C-peptide is in part a saturable process; (b) insulin clearance may be impaired in obesity and NIDDM; and (c) insulin secretion differs in obese states and NIDDM both quantitatively and qualitatively, and thereby separates the two disorders as different entities. In addition, quantitation of insulin release in obese states may also help (d) to better define primary algorithms for insulin replacement in normal- and overweight insulin-dependent diabetic patients.

Study Objective: The aim of the study was to investigate plasma concentrations of atrial natriuretic peptide, aldosterone, and renin during experimentally induced acute central venous congestion., Design: Two experimental calf models were used: (1) right heart failure due to pulmonary artery obstruction; (2) inferior vena cava syndrome produced by inferior vena caval obstruction. Hormonal responses and haemodynamic variables were measured over 6 h., Subjects: Experiments were performed on three female "Schwarzbund" calves, age 3 months, weight 92 +/- 8 kg., Measurements and Main Results: In the pulmonary artery obstructed group there was an increase of plasma aldosterone from 6.5(SEM 1.6) to 22.1(3.2) ng.dl-1 (p less than 0.05), of renin from 0.7(0.1) to 2.5(0.3) Goldblatt units x 10(-4).ml-1 (p less than 0.05), and of atrial natriuretic peptide from 22.1(4.5) to 141.4(27.8) pmol.litre-1 (p less than 0.05). During inferior vena caval obstruction, aldosterone increased from 2.4(0.4) to 20.9(2.0) ng.dl-1 (p less than 0.05), and renin increased from 0.4(0.05) to 2.0(0.20) Goldblatt units x 10(-4).ml-1 (p less than 0.05). In this experiment, atrial natriuretic peptide remained unchanged. Cardiac output decreased in both groups. There was significant fluid and electrolyte retention during both experiments, with urine volume decreasing from 87.7(11.6) to 35.0(1.2) ml-h-1 in experiment (1), and from 185(14) to 95.7(8.6) ml.h-1 in experiment (2)., Conclusions: The study suggests (1) that in an experimental acute state of reduced cardiac output due to pulmonary artery stenosis with constantly increased right heart pressures, raised endogenous atrial natriuretic peptide failed to induce diuresis and natriuresis; (2) that in acute right heart failure, renin and aldosterone secretion could not be suppressed by raised atrial natriuretic peptide concentrations; and (3) atrial natriuretic peptide secretion seemed to be exhausted after 6 h continuous atrial distension.

Hyperthyroidism is known to further impair carbohydrate metabolism in diabetic patients. In the present study we examined in noninsulin-dependent (type 2) diabetic patients the effect of T3-induced hyperthyroidism on glucose utilization and endogenous glucose production by means of the hyperinsulinemic and hyperglycemic clamp technique in combination with [3H]3-glucose kinetic analysis. Administration of T3 for 1 week increased the mean serum T3 concentration from 1.0 +/- 0.1 (SEM) to 4.1 +/- 0.2 ng/ml, and the mean fasting plasma glucose from 8.7 +/- 0.7 to 9.9 +/- 0.9 mmol/liter. Basal hepatic glucose production (HGP) rose from 3.23 +/- 0.23 to 3.98 +/- 0.25 mg/kg X min, whereas basal MCR of glucose (MCRG) increased only slightly from 2.12 +/- 0.24 to 2.30 +/- 0.14 ml/kg X min. When the plasma insulin concentration was acutely raised and maintained at 82 +/- 8 microU/ml (hyperinsulinemic clamp study), HGP decreased to 0.71 +/- 0.29 mg/kg X min and MCRG increased to 3.16 +/- 0.47 ml/kg X min. After T3 administration suppression of HGP by insulin was reduced (1.55 +/- 0.37 mg/kg X min), but at the same time MCRG was only slightly affected (3.64 +/- 0.54 ml/kg X min). In the hyperglycemic clamp study the plasma glucose concentration was maintained 7 mmol/liter above the individual fasting level. MCRG was again slightly higher after T3 administration (1.98 +/- 0.18 vs. 1.66 +/- 0.15 ml/kg X min), but insufficient to completely compensate for the higher residual HGP at the hyperthyroid as compared to the euthyroid state (2.42 +/- 0.24 vs. 1.45 +/- 0.36 mg/kg X min). Thus, deterioration of metabolic control in noninsulin-dependent diabetic patients by hyperthyroidism is due primarily to enhancement of basal HGP and its reduced suppressibility by insulin and glucose.

This study examines the feasibility of deriving the 24-h insulin requirement of insulin-dependent diabetic patients who were devoid of any endogenous insulin release (IDD) from the insulin-production rate (IPR) of healthy man (basal, 17 mU/min; stimulated 1.35 U/12.5 g glucose). To this end, continuous intravenous insulin infusion (CIVII) was initiated at a precalculated rate of 41.2 +/- 4.6 (SD) U/24 h in IDD (N - 12). Blood glucose profiles were compared with those obtained during intermittent subcutaneous (s.c.) insulin therapy (IIT) and those of healthy controls (N = 7). Regular insulin (Hoechst CS) was infused with an adapted Mill Hill Infuser at a basal infusion rate of 1.6 U/h (6:00 a.m. to 8:00 p.m.), and of 0.8 U/h from 8:00 p.m. to 6:00 a.m. Preprandial insulin (3.2-6.4 U) was added for breakfast, lunch, and dinner. Daily individual food intake totaled 7688 +/- 784 kJ (1836 +/- 187 kcal)/24 h including 184 +/- 37 g of glucose. Proper control of blood glucose (BG) (mean BG 105 +/- 10 mg/dl; mean amplitude of glycemic excursions 54 +/- 18 mg/dl; and 1 h postprandial BG levels not exceeding 160 mg/dl) and of plasma concentrations of beta-hydroxybutyrate and lactate was maintained by 41.4 +/- 4.4 U insulin/24 h. Although BG values only approximated the upper normal range as seen in healthy controls, they were well within the range reported by others during CIVII. Therefore, we conclude that in adult IDD completely devoid of endogenous insulin (1) the IPR of normal man can be used during CIVII as an estimate for the patient's minimal insulin requirement per 24 h, and (2) this approach allows for a blood glucose profile close to the upper range of a normal control group. Thus, deriving a patient's daily insulin dose from the insulin production rate of healthy man may add an additional experimental protocol which aids in making general calculations of a necessary insulin dose instead of using trial and error or a closed-loop insulin infusion system.

This preliminary study was designed to determine whether biologic action of biosynthetic human insulin (BHI) in vitro and its antigenicity in man equals that of various insulins of pancreatic origin. Testing the biologic action of BHI, pancreatic human insulin (PHI), and pancreatic pork insulin (PCPI) on glucose uptake by the rat hemidiaphragm and adipose tissue, identical efficacy for the insulins on a unit per unit basis was observed. Furthermore, a comparable impairment of insulin action was seen under the detrimental conditions of a hyperosmolal state. In an insulin-dependent diabetic female patient with cutaneous insulin hypersensitivity, BHI displayed the same antigenicity as PHI and PCPI. From these data we conclude tht BHI, both in vitro and in vivo, exhibits the same biologic properties as pancreatic insulin, and that it is reasonable to continue extensive testing of BHI in man, since identical metabolic effects and immunologic reactions are to be anticipated as with purified pancreatic insulin preparations of human or animal origin.

To determine the role of proinsulin on endogenous insulin release, splanchnic output and arterial concentrations of C-peptide were measured in healthy subjects before and during infusion of human (HPI) and porcine (PPI) proinsulin at increasing rates for 70 min each (HPI, 328 and 656 micrograms X m-2 X h-1; PPI, 54, 134, and 268 micrograms/m-2 X h-1), while euglycemia was maintained by variable glucose infusion. By using this approach splanchnic C-peptide output was reduced by human proinsulin infusion from 143 +/- 16 (mean +/- SE) pmol/min to 111 +/- 18 and 75 +/- 11 pmol/min (P = 0.01). Simultaneously, arterial concentrations of C-peptide decreased from 716 +/- 40 pmol/l by 23 and 32%. Similar inhibition was induced by porcine PPI of splanchnic C-peptide output at an infusion rate of 268 micrograms X m-2 X h-1. Mean metabolic clearance rate was 2.7 and 3.7 ml X kg-1 X min-1 for HPI and PPI, respectively. Splanchnic glucose output was almost completely suppressed by human and porcine proinsulin at maximal infusion rates. This effect preceded both inhibition by proinsulin of splanchnic C-peptide output and stimulation of peripheral glucose utilization. We conclude that human and porcine proinsulin suppress endogenous insulin secretion at pharmacological concentrations. The observed constancy of the metabolic clearance rate of HPI demonstrates that its clearance remains a nonsaturable process up to supraphysiological HPI concentrations, while clearance of PPI appears to be subject to saturation. Furthermore, it appears that splanchnic glucose output responds earlier to proinsulin exposure than suppression of C-peptide release or stimulation of peripheral glucose utilization.

To evaluate the suppressive effect of biosynthetic human insulin (BHI; 2.5 U/m2 . h) on basal and glucose-stimulated insulin secretion in healthy and obese hyperinsulinemic subjects, the plasma C-peptide response was measured during maintenance of euglycemia and hyperglycemia by means of the glucose clamp technique. In five healthy subjects in whom arterial insulin concentration was increased to 94 +/- 8 microU/mL, but euglycemia was maintained at the fasting level. C-peptide concentration fell from 1.3 +/- 1.0 ng/mL by 21 +/- 8% (P less than 0.05). When hyperglycemia of 7 mmol/L above basal was induced by a variable glucose infusion, the C-peptide response was similar in the control (5.0 +/- 0.6 ng/mL) and BHI experiments (4.7 +/- 0.6 ng/mL) and was paralleled by an identical increase in plasma insulin above the prevailing insulin concentration. In seven obese patients plasma C-peptide fell from 3.5 +/- 0.4 to 2.8 +/- 0.5 ng/mL (P less than 0.05) when BHI was infused at the same rate of euglycemia maintained as in the lean subjects. As in healthy subjects, however, the plasma C-peptide response to the hyperglycemic stimulus (8.7 +/- 0.9 ng/mL) was not altered by BHI (7.9 +/- 0.8 ng/mL). Glucose utilization as determined by the glucose infusion rate necessary to maintain the desired glucose level was reduced by half in the obese patients compared with that of normal subjects. From these data we conclude that in healthy as well as obese hyperinsulinemic subjects, insulin at concentrations capable of suppressing its basal secretion fails to suppress its glucose-stimulated secretion.

To determine the effect of the 75 g oral glucose tolerance test on carbohydrate and lipid metabolism, the splanchnic exchange of glucose, lactate, pyruvate, non-esterified fatty acids, beta-hydroxybutyrate and acetoacetate as well as the release of insulin, C-peptide, glucagon and pancreatic polypeptide were evaluated in eight healthy male volunteers in the basal state and for 150 min following glucose ingestion. Oral glucose loading was followed by a rapid rise in splanchnic output of glucose (mean +/- SEM; 154 +/- 12 mmol/150 min), pyruvate (1.2 +/- 1.2 mmol/150 min) and lactate (8.6 +/- 2.0 mmol/150 min), whereas there were reductions in the splanchnic uptake of non-esterified fatty acids (-10.7 +/- 4.4 mmol/150 min) and the splanchnic output of beta-hydroxybutyrate (-4.8 +/- 3.3 mmol/150 min) and acetoacetate (-3.0 +/- 1.2 mmol/150 min). In parallel, splanchnic output of insulin (12.3 +/- 2.7 nmol/150 min), C-peptide (36.1 +/- 5.0 nmol/150 min) and transiently of pancreatic polypeptide rose, whereas that of glucagon fell (-0.58 +/- 0.21 nmol/150 min). Even at 150 min after glucose ingestion, splanchnic output and arterial concentrations of glucose, lactate, insulin and C-peptide were still above their respective basal values while those of non-esterified fatty acids and glucagon were reduced. Taking into account the partial suppression of endogenous glucose production by ingested glucose it is concluded that, in normal postabsortive man, only 49-63% of a 75 g oral glucose load is retained by the splanchnic bed during the first 150 min, the rest being available for non-hepatic tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

To determine the impact of induced hypo- and hypercalcemia on TRH (400 micrograms)-stimulated TSH and PRL release, healthy subjects (n = 11) were infused with 5% glucose in water (n = 11), disodium EDTA (n = 11), or calcium gluconate (n = 7). TRH was given as an iv bolus 60 min (5% glucose and EDTA) and 120 min (calcium) after initiation of the respective infusion. Basal plasma concentrations of TSH remained unchanged during induced hypo- and hypercalcemia, whereas those of PRL fell during the latter (P less than 0.05). The mean sum of increments (0-90 min) in PRL and TSH was considerably greater during hypocalcemia than during hypercalcemia (PRL, P less than 0.002; TSH, P less than 0.005). The increments in the plasma hormone concentration above basal after iv TRH were increased compared to those in normocalcemia (PRL, 98.4 +/- 37.9 ng/ml; TSH, 38.9 +/- 11.8 microU/ml) during hypocalcemia [PRL, 128 +/- 47.8 ng/ml (P less than 0.002); TSH, 46.7 +/- 12.8 microU/ml; (P less than 0.005)], but were impaired during hypercalcemia [PRL, 70.1 +/- 27 ng/ml (P less than 0.002); TSH, 28.9 +/- 8.5 microU/ml (P less than 0.025)]. The mean sum of increments in PRL was related to concentrations of both serum calcium (r = -0.59; P less than 0.01) and PTH (r = 0.51; P less than 0.05). A relation was also seen between the incremental responses of TSH and serum calcium (r = -0.52; P less than 0.05), PTH (r = 0.55; P less than 0.01), and phosphorus (r = -0.55; P less than 0.01). We conclude that in healthy man, TRH-mediated release of both PRL and TSH are inversely related to serum calcium concentrations in such a manner that hormone secretion is enhanced by acute hypocalcemia, but blunted by hypercalcemia.

To examine the effect of experimental hyperthyroidism induced by triiodothyronine (T3) administration on glucose utilization and glucose-stimulated insulin secretion, euglycemic insulin clamp studies (A), hyperglycemic clamp studies (B), and estimation of splanchnic glucose metabolism after an oral glucose load by means of the hepatic venous catheter technique (C) were performed in healthy volunteers whose serum T3 concentration had been increased from 1.1 +/- 0.02 (SE) to 3.5 +/- 0.2 ng/ml. At plasma insulin concentrations of 77 +/- 6 microU/ml and maintenance of euglycemia, insulin-mediated glucose metabolism (6.8 +/- 0.4 mg X kg-1 X min-1) was not significantly altered following T3 exposure (7.2 +/- 0.7) in study A. When plasma glucose was raised and maintained at 7 mmol/l above basal, glucose uptake (10.6 +/- 0.7 mg X kg-1 X min-1) was increased to 14.6 +/- 2.0 mg X kg-1 X min-1 (P less than 0.02) following T3 administration in study B. In study C, basal splanchnic glucose output (104 +/- 10 mg/min) was markedly augmented when preceded by T3 exposure (167 +/- 17 mg/min; P less than 0.005). After a 75-g glucose load, splanchnic glucose output was enhanced by T3 by 30% (43.4 +/- 2.2 vs. 56.9 +/- 6.6 g/150 min; P less than 0.05), whereas arterial plasma concentrations of glucose, insulin, and C-peptide were not significantly different. It is concluded that in healthy humans, exposure to elevated plasma concentrations of T3 increases the amount of glucose to be released from the splanchnic bed. Glucose tolerance, however, remains unaltered, because peripheral glucose uptake is increased at the same time.

Rioprostil is a synthetic prostaglandin E1 analogue currently under study for peptic ulcer healing. Rioprostil, like prostaglandin E, caused an impairment of glucose tolerance in animal studies. We have studied the effects of single therapeutic doses of rioprostil (300 micrograms and 600 micrograms) on glucose homeostasis after oral glucose tolerance tests in 12 healthy volunteers. Plasma glucose, insulin, and C-peptide concentrations were not significantly different after rioprostil and placebo. These results indicate that a clinically significant interference of rioprostil with glucose tolerance is unlikely.

In 5 total artificial heart (TAH)-bridge-to-transplant (BTT) patients production and secretion of atrial natriuretic factor (ANF)1 was studied before, during, and after TAH implantation. Bridging periods lasted between 9 and 28 days. Atrial biopsies were taken during implantation, and after TAH explantation for histologic and histochemical investigations to evaluate differences in morphology and ANF-content of the specific atrial granules. Plasma concentrations of ANF (pANF), aldosterone (PAC) and renin (PRC) were measured daily, as were hemodynamic parameters. In the preoperative state, pANF was always markedly elevated, while during TAH bridging, pANF remained moderately elevated with fluctuations. A positive correlation between pANF levels and right atrial pressure (RAP) was seen in all patients (p less than 0.05). Slight correlation was also observed between pANF and left atrial pressure (LAP), but no correlation was seen between pANF and systemic blood pressure, and no consistent pattern was seen in the plasma concentrations of either renin or aldosterone. After heart transplantation (HTX), which was performed in 3 patients, pANF levels were significantly higher than during TAH, and continued to show a positive correlation with RAP. From our data, we conclude that ANF production sites and secretory mechanisms remain intact during TAH-bridging, although upon implantation of a TAH, the remaining atria are deprived of all coronary blood supply and most autonomic innervation.

To evaluate the role of pulsatile insulin administration, hepatic glucose production (HGP) and utilization were studied in type I diabetic patients in the fasting state and during a euglycemic insulin (1 mU X kg-1 X min-1 i.v.) clamp with continuous and pulsatile insulin administration. In the latter study, insulin was infused at twice the continuous rate with 3-min-on/7-min-off intervals, thereby reducing total insulin delivery by 40%. The restraining effect of pulsatile insulin on basal HGP (1.91 +/- 0.35 mg X kg-1 X min-1) was equipotent to continuous insulin exposure (1.80 +/- 0.17 mg X kg-1 X min-1). During the insulin-clamp studies, HGP was equally suppressed by pulsed (0.62 +/- 0.12 mg X kg-1 X min-1) as by continuous insulin infusion (0.63 +/- 0.12 mg X kg-1 X min-1). Insulin-stimulated glucose utilization was not significantly altered in either study (2.55 +/- 0.27 vs. 2.92 +/- 0.23 mg X kg-1 X min-1). When in further studies the total insulin dose given during the pulsatile study was infused continuously (0.6 mU X kg-1 X min-1), HGP in the basal state and residual HGP during the insulin-clamp study were 25-30% higher than in the pulsatile experiments, whereas glucose utilization was not significantly different. In conclusion, by reducing total hormone delivery by up to 40%, but given in a pulsatile fashion, insulin is equally potent in controlling HGP as continuous insulin administration. This greater efficacy of pulsatile exposure in suppressing HGP is accompanied by an equipotent effect on glucose utilization.(ABSTRACT TRUNCATED AT 250 WORDS)

To study the interdependence of utilization of branched chain amino acids and glucose and of hyperinsulinaemia in patients with liver cirrhosis the plasma disappearance of glucose and amino acids was estimated in seven patients with cirrhosis and portocaval shunt and in seven healthy controls following infusion of glucose and essential amino acids during suppression of endogenous hormone release by somatostatin. Exogenous insulin was infused by means of an automated glucose controlled insulin infusion system. The data demonstrate that (i) insulin requirement almost doubled in patients as compared to controls to obtain similar blood glucose responses to i.v. glucose, and (ii) the plasma disappearance rates of the infused amino acids were reduced in the patients as compared to controls despite hyperinsulinaemia sufficient to achieve normal glucose assimilation. Thus, in cirrhotic patients insulin resistance may be overcome by excess insulin as far as glucose homoestasis is concerned, whereas amino-acid metabolism still remains impaired.

This is a case report of a 43 years old phaenotypic female (karyotype 46 XY) with congenital adrenal hyperplasia and male pseudohermaphroditism presenting with severe hypertension and hypokalaemic alkalosis. Evaluation of the steroid biosynthesis of the adrenals as well as of the intraabdominal testes demonstrated a severe degree of 17-alpha-hydroxylase deficiency in this patient. This defect was associated by a complete lack of deoxycortisol and cortisol as well as of testicular testosterone synthesis, and by an overproduction of DOC and corticosterone. Sruvival of the patient in spite of severe cortisol deficiency was due to the glucocorticoid activity of corticosterone. This compound and DOC account also -- due to their mineralocorticoid properties -- for the hypertensive state of this male pseudohermaphrodite.

Elevated plasma concentrations of growth hormone impair glucose tolerance and insulin sensitivity of peripheral tissues. To study the effect of short-term exposure to growth hormone concentrations elevated into the upper physiologic range (7-10 ng/ml) on splanchnic carbohydrate metabolism, both splanchnic glucose output (SGO) and substrate exchange after ingestion of a 75-g glucose load were determined by means of the liver vein catheter technique in six healthy volunteers after growth hormone administration. Growth hormone was infused at a rate of 2 micrograms/kg X h starting 120 min before and continuing for 150 min following the glucose load. Control studies without growth hormone administration were performed in seven subjects. SGO was 104 +/- 10 (SEM) mg/min in the postabsorptive state and increased to 43.4 +/- 2.2 g during the 150-min period following glucose ingestion. Growth hormone infusion did not alter basal SGO (130 +/- 14 mg/min), nor the splanchnic exchange of lactate, pyruvate, and free fatty acids, whereas basal production of beta-OH-butyrate was increased twofold; following glucose ingestion a higher proportion of the given glucose load escaped the splanchnic bed after growth hormone exposure (66.9 +/- 6.8 g/150 min; P less than 0.005). The insulin production rate (basal 14 +/- 2 mU/min; following oral glucose 7.0 +/- 0.8 U/150 min) as calculated from C-peptide release from the splanchnic area was unaltered by growth hormone exposure in the basal state (14 +/- 3 mU/min), but augmented after glucose ingestion (14.8 +/- 1.5 U/150 min).(ABSTRACT TRUNCATED AT 250 WORDS)

To examine the fate of insulin across the liver bed, biosynthetic human insulin was infused in increasing amounts in six healthy men. Trapping of insulin by the liver was determined by means of the hepatic venous catheter technique. To minimize any possible error in the estimation of insulin removal as a result of endogenous insulin, pancreatic insulin secretion was suppressed by intravenous administration of somatostatin (500 micrograms/h). Infusion rates of human insulin were 30, 60, and 150 pmol/m2 X min (corresponding to 0.25, 0.5, and 1.25 U/m2 X h) for 70 minutes each. Steady-state insulin levels were within the physiologic range, ie, 69 +/- 2, 135 +/- 3, and 342 +/- 10 pmol/L, respectively. Euglycemia was maintained throughout the study by a variable glucose infusion. The output of C-peptide from the splanchnic bed was reduced by somatostatin by about 90%, indicating that endogenous insulin production only minimally contributed to total insulin levels achieved during infusion of exogenous human insulin. Fractional extraction of insulin by the liver (63 +/- 6%, 71 +/- 4%, and 74 +/- 5%) and hepatic insulin clearance (201 +/- 19, 235 +/- 23, and 245 +/- 29 mL/m2 X min) did not differ significantly during the three insulin infusion studies. The hepatic uptake of insulin rose with increasing insulin infusion rates, constituting 40% to 60% of total-body insulin removal. No change in the total metabolic clearance rate of insulin was observed between the groups (451 +/- 8 mL/m2 X min). To study the extraction rate of C-peptide by the liver, porcine C-peptide was also infused at the same increasing rates.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of buformin (100 mg b.i.d. for 5 days) on carbohydrate metabolism, both splanchnic glucose output (SGO) and net substrate exchange were studied in 6 healthy male volunteers in the basal state and following glucose ingestion (100 g). Control studies without buformin were also performed in 5 men. Splanchnic glucose and substrate exchange was determined by means of the hepatic venous catheter technique. SGO was 154 +/- 18 (SEM) mg/min in the postabsorptive state and increased 33.3 +/- 2.8 g above the basal level during the 150 min period following glucose ingestion. Buformin administration did not alter basal SGO (157 +/- 26 mg/min), nor the splanchnic exchange of pyruvate, alanine, glycerol, OH-butyrate and acetoacetate. Splanchnic lactate balance was altered by buformin and net lactate output occurred. Following glucose ingestion the rise in splanchnic lactate output was increased, whereas no change in SGO (32.9 +/- 3.5 g/150 min) and splanchnic exchange of the other substrates was observed. The increase in arterial blood glucose concentration following oral glucose loading was reduced by buformin pretreatment (p less than 0.005). The insulin production rate (basal, 16 +/- 2 mU/min; following oral glucose, 13 +/- 2 U/150 min) as calculated from C-peptide release from the splanchnic area was unchanged by buformin. Except for a marked rise in splanchnic lactate production, buformin did not alter splanchnic carbohydrate metabolism after orally ingested glucose in healthy man. The diminished increase in arterial blood glucose concentration associated with unaltered insulin production suggests that buformin facilitates glucose utilization by peripheral tissues.

To determine the impact of biosynthetic human insulin (BHI) on endogenous insulin release, splanchnic output and arterial concentrations of C-peptide were measured in eight healthy men after intravenous administration of 0, 0.5, 1.25, U BHI . m-2 . h-1 for 70 min each. Euglycemia was maintained by a variable glucose infusion. Arterial levels of serum insulin were 48 +/- 6 pmol/liter before and 135 +/- 12, 265 +/- 18, and 593 +/- 47 pmol/liter after BHI infusion. Splanchnic C-peptide output was reduced by BHI infusion from 88 +/- 10 pmol/min before to 50 +/- 9, 28 +/- 10, and 18 +/- 16 pmol/min (P less than 0.0025). Simultaneously, arterial concentrations of C-peptide fell from 539 +/- 54 pmol/liter by 29 and 43% when 1.25 and 2.5 U . m-2 . h-1 of BHI were administered. Hepatic insulin uptake was directly related with BHI infusion rate (r = 0.88) and rose during BHI administration from a basal value of 58 +/- 7 to an uptake of 265 +/- 31 pmol/min when 2.5 U . m-2 . h-1 were infused (P less than 0.0005). Basal hepatic insulin clearance was 4.75 +/- 0.60 ml . kg-1 . min-1 and remained unchanged after BHI infusion as did hepatic fractional extraction of insulin, which was 61 +/- 4% in the basal state. Metabolic clearance rate of immunoreactive insulin (MCRi) was dose-dependently reduced by BHI infusion, whereas the relative share of hepatic insulin clearance in total MCRi rose simultaneously (P less than 0.01). We conclude that feedback inhibition of endogenous insulin release may play an important role in vivo. Furthermore, it appears that nonhepatic insulin degradation is a saturable phenomenon as total MCRi fell in the presence of its unchanged hepatic clearance rate after the infusion of large amounts of BHI.

To determine the impact of insulin-binding antibodies on total (TIRI) and free insulin (FIRI) as well as on insulin sensitivity, 10 insulin-dependent diabetic patients (IDDM) with poststimulatory C-peptide less than 100 pmol/L and an insulin binding capacity (IBC) between less than 1 and 294 micrograms/L serum were studied during and after a 1-h nonprimed, constant-rate insulin infusion (study 1: 0.057 U/kg body wt, study 2: 0.286 U/kg body wt). Euglycemia was maintained by variable glucose infusion. Control studies were performed in 5 healthy subjects. Basal TIRI (mU/L) was lowest in healthy subjects (16 +/- 1 [SE]) and elevated in diabetic patients (IBC less than 25 micrograms/L: 72 +/- 11, IBC greater than 25 micrograms/L: 1772 +/- 842), whereas serum concentrations of FIRI were considerably smaller but still two- to threefold greater (P less than 0.01) in the patients than in healthy subjects (13 +/- 1). After intravenous (i.v.) insulin administration, almost identical increments in serum TIRI were seen in healthy subjects and in diabetic patients with low IBC (less than 25 micrograms/L), whereas those with high IBC (greater than 25 micrograms/L) had a heterogeneous response.(ABSTRACT TRUNCATED AT 250 WORDS)

To compare cortisol and epinephrine action on oral glucose tolerance, healthy humans were infused with either cortisol (0.1 mg X kg-1 X h-1), epinephrine (5.4 micrograms X kg-1 X h-1), or saline before and after a 75-g glucose load, thereby elevating the respective plasma hormone concentrations into the pathophysiologic range. In the basal state, epinephrine increased arterial concentrations of glucose, beta-hydroxybutyrate, and free fatty acids (FFA) as well as splanchnic output of glucose and beta-hydroxybutyrate and splanchnic FFA more than cortisol. Postprandially, C-peptide release and hyperinsulinemia were blunted by epinephrine initially and increased less thereafter than during cortisol infusion. The rise in arterial glucose after glucose ingestion as calculated by the area under the curve was more marked (P less than .01) after epinephrine [( 1.90 +/- 0.08 M) 150 min] and cortisol [( 1.41 +/- 0.05 M) 150 min] than in the control study [( 1.07 +/- 0.04 M) 150 min]. In parallel, the stress hormones induced an almost identical 24 and 31% rise in mean splanchnic glucose output versus control values (normal, 44.8 +/- 2.5; cortisol, 55.3 +/- 3.3; epinephrine, 58.9 +/- 6.9 g/150 min). The associated rise in arterial concentrations and splanchnic output of insulin above control values was considerably greater during cortisol but unchanged during epinephrine exposure. Epinephrine but not cortisol induced a rise versus the control study in splanchnic uptake of lactate and FFA, as well as in pyruvate output, whereas plasma beta-hydroxybutyrate and acetoacetate remained unchanged. The postprandial splanchnic glucose output-to-splanchnic C-peptide output ratio did not differ from normal during epinephrine but was reduced (P less than .01) during cortisol administration.(ABSTRACT TRUNCATED AT 250 WORDS)

To determine the pharmacokinetics of insulin administered by intravenous (IV) and subcutaneous (SC) pump treatment as well as by the conventional subcutaneous route, six insulin preparations of either porcine or human amino acid sequence were investigated intraindividually following IV or SC insulin infusion at two different rates (Study I) and three preparations were investigated after SC bolus injection (Study II) in healthy men. Insulin release was suppressed in Study I by IV administration of somatostatin (500 micrograms/hr) to avoid interference by endogenous insulin with the measurement of exogenous insulin. Hypoglycemia was prevented by IV administration of glucose. The data obtained demonstrated (1) greater serum concentrations of immunoreactive insulin (IRI) during continuous IV insulin infusion (141 +/- 10 (SEM) pmole/liter) than during SC insulin infusion (54 +/- 3 pmole/liter; P less than 0.0005) (0.8 U/hr); (2) return of serum IRI to baseline values following a 17-minute square wave insulin infusion (12.8 U/hr; time: 0 to 17 minutes) within 40 minutes after IV insulin infusion but not before 180 minutes after the end of SC insulin infusion; (3) peak serum IRI at 60 to 90 minutes after conventional SC insulin injection returning to baseline values at 300 minutes; and (4) identity of the pharmacokinetics of pumped human and porcine insulin within a given group as well as of the accompanying metabolic dynamics of blood glucose and nonesterified fatty acids, but heterogeneity of serum insulin after its SC bolus injection. We conclude that (1) the pharmacokinetic behavior of regular insulin depends primarily on its route of administration; (2) continuous IV infusion of an insulin dose causes significantly higher serum insulin levels than the SC administration of the identical insulin dose, and (3) hyperinsulinemia caused by a square wave insulin infusion (12.8 U/hr; time: 0 to 17 minutes) requires more than four times longer to return to baseline levels following SC administration than after IV administration of the insulin. These differences in the pharmacokinetic behavior of insulin cause a reduced bioavailability of SC administered insulin and have to be taken into account when instituting insulin treatment by various routes.

Pharmacokinetic methods are a powerful tool for the investigation of the insulin system in health and disease; the underlying formalisms are simple and straight-foreward. Following intravenous injection the plasma concentration of insulin declines with at least two exponentials, the rapidly disappearing component (half-time 2.4 min) represents elimination from the intravascular space, whereas the most slowly disappearing component (half-time 50-130 min) reflects elimination from the interstitial fluid and the tissues that utilize insulin. Total clearance of insulin, which is the result of metabolic degradation, ranges between 700 and 800 ml/min. The sites of degradation are primarily the liver (hepatic insulin clearance: 320-400 ml/min) and the kidneys (renal insulin clearance: 190-270 ml/min), hence insulin disposal depends on the function of these organs. The apparent volume of distribution for insulin is approximately equal to the extracellular space. Insulin absorption from the subcutaneous tissue is slow (half-times of 0.5-2.7, 6.6-13.8, and 15-48 hours for fast-, intermediate-, and long-acting insulins, respectively). The bioavailability of insulin after subcutaneous administration is variable. Insulin kinetics appear not to be altered in diabetes mellitus, except in cases with insulin antibodies or in insulin-resistant patients, in which insulin removal may be retarded. Optimization of functional insulin replacement requires knowledge of the pharmacokinetic properties of the insulin preparation used.

The effect of acetyl-salicylic acid (ASA, 3 g per day for 3 days) on glucose utilization and insulin secretion was studied in healthy volunteers and Type 2 diabetic patients using the hyperglycaemic and euglycaemic insulin clamp technique. When in healthy subjects arterial plasma glucose was acutely raised and maintained at +7 mmol/l above fasting level, the plasma insulin response was enhanced by ASA (70 +/- 7 vs. 52 +/- 7 mU/l), whereas the plasma C-peptide response was identical. Despite higher insulin concentrations, glucose utilization was not significantly altered (control, 61 +/- 7; ASA, 65 +/- 6 mumol X kg-1 X min-1) indicating impairment of tissue sensitivity to insulin by ASA. Inhibition of prostaglandin synthesis was not likely to be involved in the effect of ASA, since insulin response and glucose utilization were unchanged following treatment with indomethacin. In the euglycaemic insulin (1 mU X kg-1 X min-1) clamp studies, glucose utilization was unaltered by ASA despite higher insulin concentrations achieved during constant insulin infusion (103 +/- 4 vs. 89 +/- 4 mU/l). In Type 2 diabetic patients, fasting hyperglycaemia (10.6 +/- 1.1 mmol/l) and hepatic glucose production (15 +/- 2 mumol X kg-1 X min-1) fell upon ASA treatment (8.6 +/- 0.7 mmol/l; 13 +/- 1 mumol X kg-1 X min-1). During the hyperglycaemic clamp study, the plasma response of insulin, but not of C-peptide, was enhanced by ASA, whereas tissue sensitivity to insulin was reduced by 30 percent.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma concentrations of proteins secreted by the liver (prealbumin, haptoglobin, transferrin, ceruloplasmin, alpha 1-antitrypsin, antithrombin III, and T4-binding globulin) and proteins mainly derived from endothelium [fibronectin, angiotensin-converting enzyme (ACE), and factor VIII-related antigen (F VIII R:Ag)] were measured in 27 hyperthyroid and 30 normal women. Significantly increased plasma concentrations (P less than 0.01) of endothelium-associated proteins, including fibronectin, ACE, and F VIII R:Ag, were found in hyperthyroid patients, while levels of proteins of primarily hepatic origin were normal. To determine whether the increase in endothelium-associated proteins in hyperthyroidism was directly related to elevated thyroid hormone levels, seven normal women were given T3 (25 micrograms, three times daily) for 2 weeks. These women had a consistent rise (P less than 0.05) in plasma concentrations of fibronectin, ACE, F VIII R:Ag, and tissue plasminogen activator. The rise in endothelium-associated proteins persisted for 10 days after cessation of T3. Plasma concentrations of hepatically synthesized proteins did not change. We conclude that thyroid hormone action either promotes endothelial protein synthesis or impairs its clearance.