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Abstract Background Hepatic insulin clearance (HIC) is an important pathophysiology of type 2 diabetes mellitus (T2DM). HIC was reported to decrease in patients with type 2 diabetes and metabolic syndrome. HIC is originally calculated by post-load insulin and C-peptide from the oral glucose tolerance test (OGTT). However, OGTT or meal tolerance tests are a burden for patients, and OGTT is not suitable for overt diabetes due to the risk of hyperglycemia. If we can calculate the HIC from the fasting state, it is preferable. We hypothesized that fasting HIC correlates with postprandial HIC in both participants with T2DM and without diabetes. We investigated whether fasting HIC correlates with postprandial HIC in overt T2DM and nondiabetes subjects (non-DM) evaluated by using glucose clamp and meal load. Methods We performed a meal tolerance test and hyperinsulinemic–euglycemic clamp in 70 subjects, 31 patients with T2DM and 39 non-DM subjects. We calculated the postprandial C-peptide AUC-to-insulin AUC ratio as the postprandial HIC and the fasting C-peptide-to-insulin ratio as the fasting HIC. We also calculated whole-body insulin clearance from the glucose clamp test. Results The fasting HIC significantly correlated with postprandial HIC in T2DM (r_S = 0.82, P

Background: Hepatic insulin clearance (HIC) is an important pathophysiology of type 2 diabetes mellitus (T2DM). HIC was reported to decrease in patients with type 2 diabetes and metabolic syndrome. HIC is originally calculated by post-load insulin and C-peptide from the oral glucose tolerance test (OGTT). However, OGTT or meal tolerance tests are a burden for patients, and OGTT is not suitable for overt diabetes due to the risk of hyperglycemia. If we can calculate the HIC from the fasting state, it is preferable. We hypothesized that fasting HIC correlates with postprandial HIC in both participants with T2DM and without diabetes. We investigated whether fasting HIC correlates with postprandial HIC in overt T2DM and nondiabetes subjects (non-DM) evaluated by using glucose clamp and meal load. Methods: We performed a meal tolerance test and hyperinsulinemic–euglycemic clamp in 70 subjects, 31 patients with T2DM and 39 non-DM subjects. We calculated the postprandial C-peptide AUC-to-insulin AUC ratio as the postprandial HIC and the fasting C-peptide-to-insulin ratio as the fasting HIC. We also calculated whole-body insulin clearance from the glucose clamp test. Results: The fasting HIC significantly correlated with postprandial HIC in T2DM (r_S = 0.82, P < 0.001). Nondiabetes subjects also showed a significant correlation between fasting and postprandial HIC (r_S = 0.71, P < 0.001). Fasting HIC in T2DM was correlated with BMI, HbA1c, gamma-glutamyl transpeptidase, HOMA-IR, HOMA-beta, M/I, and whole-body insulin clearance. Fasting HIC in nondiabetes subjects was correlated with HOMA-IR and HOMA-beta. Conclusions: These results suggest that fasting HIC is strongly correlated with postprandial HIC in both overt T2DM and non-DM patients, as evaluated by the meal test and glucose clamp method. Fasting HIC could be a convenient marker of HIC. [ABSTRACT FROM AUTHOR]

Mammals are protected from changes in environmental temperature by altering energetic processes that modify heat production. Insulin is the dominant stimulus of glucose uptake and metabolism, which are fundamental for thermogenic processes. The purpose of this work was to determine the interaction of ambient temperature induced changes in energy expenditure (EE) on the insulin sensitivity of glucose fluxes. Short-term and adaptive responses to thermoneutral temperature (TN, ∼28 °C) and room (laboratory) temperature (RT, ∼22 °C) were studied in mice. This range of temperature does not cause detectable changes in circulating catecholamines or shivering and postabsorptive glucose homeostasis is maintained. We tested the hypothesis that a decrease in EE that occurs with TN causes insulin resistance and that this reduction in insulin action and EE is reversed upon short term (

Background: Fasting insulin resistance indexes are used extensively nowadays. We intended to analyze a new recently presented fasting index, SPISE (sensitivity formula: 600 × HDL-cholesterol0.185/triglycerides0.2/BMI1.338), in comparison with three previously known fasting indexes, regarding correlation with the insulin clamp index, and for the predictive effects of future long-term risks of coronary heart disease (CHD) or manifest type 2 diabetes. Methods: A total of 1049 71-year-old male subjects from the Swedish ULSAM study, median follow-up 8 years, were included. All subjects performed the euglycemic insulin clamp, and analyses of four fasting insulin resistance indexes: SPISE-IR (= 10/SPISE), QUICKI-IR, Log HOMA-IR, and Revised QUICKI-IR. Results: Spearman correlation coefficients with the insulin clamp were 0.60–0.62 for all indexes. Area under curve at ROC analysis was 0.80 for SPISE-IR, and 0.84 for QUICKI-IR, Log HOMA-IR, and Rev QUICKI-IR. Adjusted hazard ratios per 1 SD index increase for long-term risk CHD were similar in all patients: 1.20–1.24 (p = 0.02–0.03). However, comparing the highest quartile (recommended to define insulin resistance) with the lower quartiles, SPISE-IR was the strongest and the only statistically significant insulin resistance index: HR 1.53 (p = 0.02). Adjusted odds ratios per 1 SD index increase for long-term risk of type 2 diabetes were fairly similar (p

Objective: Increased deposition of the extracellular matrix (ECM) in adipose tissue (AT) during obesity contributes to insulin resistance. The integrin receptors transmit changes in the extracellular environment causing corresponding intracellular adaptations. Integrin-linked kinase (ILK), an adaptor protein, is a central hub for intracellular signaling of integrins. This study determined the role of ILK in adipose function and insulin resistance. Methods: The pathogenic role of ILK in obesity and insulin resistance was studied in human adipose tissue and adipocyte-specific ILK-deficient mice (ILKlox/loxAdCre). ILKlox/loxAdCre mice together with wild-type littermates (ILKlox/lox) were fed a chow diet or 60% high-fat (HF) diet for 16 weeks. In vivo insulin sensitivity was determined by hyperinsulinemic-euglycemic clamps. Results: AT ILK expression was increased by HF diet feeding in mice and increased in visceral fat of morbidly obese humans. The HF-fed ILKlox/loxAdCre mice displayed reduced fat mass and improved glucose tolerance relative to the HF-fed ILKlox/lox mice. During a hyperinsulinemic-euglycemic clamp, the HF-fed ILKlox/loxAdCre mice exhibited partially improved insulin resistance in AT. Lipolysis was suppressed to a greater extent by insulin and glucose uptake in brown AT (BAT) increased. Increased inhibition of lipolysis may have been attributed to increased vascularization in white AT, while increased glucose uptake in BAT was associated with increased Akt phosphorylation and P38/JNK dephosphorylation. Notably, AT insulin sensitivity in lean mice was not affected by ILK deletion. Moreover, reduced fat mass in the HF-fed ILKlox/loxAdCre mice may have been attributed to decreased free fatty acid uptake into adipocytes via the downregulation of CD36 gene expression. Consistent with the results in the mice, knockdown and knockout of ILK in 3T3-L1 cells decreased lipid accumulation and CD36 gene expression during adipogenesis. Conclusions: These data show that adipocyte ILK is important for regulating HF diet-mediated insulin resistance in AT in a manner consistent with AT function.

Purpose: Skeletal muscle is the primary site for insulin-stimulated glucose disposal, and muscle insulin resistance is central to abnormal glucose metabolism in obesity. Whether muscle insulin signaling to the level of Akt/AS160 is intact in insulin-resistant obese humans is controversial.Methods: We defined a linear range of insulin-stimulated systemic and leg glucose uptake in 14 obese and 14 nonobese volunteers using a 2-step insulin clamp (Protocol 1) and then examined the obesity-related defects in muscle insulin action in 16 nonobese and 25 obese male and female volunteers matched for fitness using a 1-step, hyperinsulinemic, euglycemic clamp coupled with muscle biopsies (Protocol 2).Results: Insulin-stimulated glucose disposal (Si) was reduced by > 60% (P < 0.0001) in the obese group in Protocol 2; however, the phosphorylation of Akt and its downstream effector AS160 were not different between nonobese and obese groups. The increase in phosphorylation of Akt2 in response to insulin was positively correlated with Si for both the nonobese (r = 0.53, P = 0.03) and the obese (r = 0.55, P = 0.01) groups. Total muscle GLUT4 protein was 17% less (P < 0.05) in obese subjects.Conclusions: We suggest that reduced muscle glucose uptake in obesity is not due to defects in the insulin signaling pathway at the level of Akt/AS160, which suggests there remain significant gaps in our knowledge of muscle insulin resistance in obesity. Our data imply that models of acute lipotoxicity do not replicate the pathophysiology of obesity. [ABSTRACT FROM AUTHOR]

Mammals are protected from changes in environmental temperature by altering energetic processes that modify heat production. Insulin is the dominant stimulus of glucose uptake and metabolism, which are fundamental for thermogenic processes. The purpose of this work was to determine the interaction of ambient temperature induced changes in energy expenditure (EE) on the insulin sensitivity of glucose fluxes. Short-term and adaptive responses to thermoneutral temperature (TN, ∼28 °C) and room (laboratory) temperature (RT, ∼22 °C) were studied in mice. This range of temperature does not cause detectable changes in circulating catecholamines or shivering and postabsorptive glucose homeostasis is maintained. We tested the hypothesis that a decrease in EE that occurs with TN causes insulin resistance and that this reduction in insulin action and EE is reversed upon short term (<12h) transition to RT. Insulin-stimulated glucose disposal (Rd) and tissue-specific glucose metabolic index were assessed combining isotopic tracers with hyperinsulinemic-euglycemic clamps. EE and insulin-stimulated Rd are both decreased (∼50%) in TN-adapted vs RT-adapted mice. When RT-adapted mice are switched to TN, EE rapidly decreases and Rd is reduced by ∼50%. TN-adapted mice switched to RT exhibit a rapid increase in EE, but whole-body insulin-stimulated Rd remains at the low rates of TN-adapted mice. In contrast, whole body glycolytic flux rose with EE. This higher EE occurs without increasing glucose uptake from the blood, but rather by diverting glucose from glucose storage to glycolysis. In addition to adaptations in insulin action, 'insulin-independent' glucose uptake in brown fat is exquisitely sensitive to thermoregulation. These results show that insulin action adjusts to non-stressful changes in ambient temperature to contribute to the support of body temperature homeostasis without compromising glucose homeostasis. • EE and insulin-mediated glucose fluxes are reduced in thermoneutral(TN)-adapted vs room 'laboratory' temperature (RT) mice. • Reduced insulin sensitivity manifests in TN mice whether they are TN-adapted or short-term transitioned from RT-adapted to TN. • TN-adapted mice are resistant to RT-induced increase in insulin sensitivity even though metabolic rate is increased. • TN-adapted mice switched to RT meets increased thermogenic needs by partitioning glucose from glycogen storage to glycolysis. • Brown fat glucose uptake sensitively increases with RT and decreases with TN by an insulin-independent mechanism. [ABSTRACT FROM AUTHOR]

Background: Fasting insulin resistance indexes are used extensively nowadays. We intended to analyze a new recently presented fasting index, SPISE (sensitivity formula: 600 × HDL-cholesterol0.185/triglycerides0.2/BMI1.338), in comparison with three previously known fasting indexes, regarding correlation with the insulin clamp index, and for the predictive effects of future long-term risks of coronary heart disease (CHD) or manifest type 2 diabetes. Methods: A total of 1049 71-year-old male subjects from the Swedish ULSAM study, median follow-up 8 years, were included. All subjects performed the euglycemic insulin clamp, and analyses of four fasting insulin resistance indexes: SPISE-IR (= 10/SPISE), QUICKI-IR, Log HOMA-IR, and Revised QUICKI-IR. Results: Spearman correlation coefficients with the insulin clamp were 0.60–0.62 for all indexes. Area under curve at ROC analysis was 0.80 for SPISE-IR, and 0.84 for QUICKI-IR, Log HOMA-IR, and Rev QUICKI-IR. Adjusted hazard ratios per 1 SD index increase for long-term risk CHD were similar in all patients: 1.20–1.24 (p = 0.02–0.03). However, comparing the highest quartile (recommended to define insulin resistance) with the lower quartiles, SPISE-IR was the strongest and the only statistically significant insulin resistance index: HR 1.53 (p = 0.02). Adjusted odds ratios per 1 SD index increase for long-term risk of type 2 diabetes were fairly similar (p < 0.001) in all patients: 1.62 for SPISE-IR, 1.97 for QUICKI-IR and Log HOMA-IR, and 2.04 for Rev QUICKI-IR, and also when comparing the highest versus the lower quartiles: 2.8–3.1 (p < 0.001). Conclusion: SPISE, easily applicable, performed equally well as other fasting insulin indexes previously recommended for clinical use, regarding correlation with the insulin clamp, and as predictor for future long-term risks of CHD or type 2 diabetes. [ABSTRACT FROM AUTHOR]

Context: Little is known about the acute effects of resistance exercise on insulin sensitivity in people with type 1 diabetes. Design: Repeated-measures design with 2 independent variables: group (exercise and nonexercise control) and time (preexercise and 12 and 36 h postexercise). Setting: General Clinical Research Center, Temple University Hospital, Philadelphia, PA. Patients: 14 physically active subjects (11 men and 3 women) with type 1 diabetes. Intervention: The exercise group completed 5 sets of 6 repetitions of strenuous (80% 1-RM) quadriceps and hamstring exercises while the control group performed only activities of daily living. Main Outcome Measures: Insulin sensitivity was assessed with the euglycemic-hyperinsulinemic-clamp technique preexercise and 12 and 36 h postexercise. Results: Insulin-sensitivity values were not significantly different between the exercise and control groups (P = .92) or over time (P = .67). Conclusions: A single bout of strenuous resistance exercise does not alter insulin sensitivity in people with type 1 diabetes. [ABSTRACT FROM AUTHOR]

Abstract Objective Appropriate glucose levels are essential for survival; thus, the detection and correction of low blood glucose is of paramount importance. Hypoglycemia prompts an integrated response involving reduction in insulin release and secretion of key counter-regulatory hormones glucagon and epinephrine that together promote endogenous glucose production to restore normoglycemia. However, specifically how this response is orchestrated remains to be fully clarified. The low affinity hexokinase glucokinase is found in glucose-sensing cells involved in glucose homeostasis including pancreatic β-cells and in certain brain areas. Here, we aimed to examine the role of glucokinase in triggering counter-regulatory hormonal responses to hypoglycemia, hypothesizing that reduced glucokinase activity would lead to increased and/or earlier triggering of responses. Methods Hyperinsulinemic glucose clamps were performed to examine counter-regulatory responses to controlled hypoglycemic challenges created in humans with monogenic diabetes resulting from heterozygous glucokinase mutations (GCK-MODY). To examine the relative importance of glucokinase in different sensing areas, we then examined responses to clamped hypoglycemia in mice with molecularly defined disruption of whole body and/or brain glucokinase. Results GCK-MODY patients displayed increased and earlier glucagon responses during hypoglycemia compared with a group of glycemia-matched patients with type 2 diabetes. Consistent with this, glucagon responses to hypoglycemia were also increased in I366F mice with mutated glucokinase and in streptozotocin-treated β-cell ablated diabetic I366F mice. Glucagon responses were normal in conditional brain glucokinase-knockout mice, suggesting that glucagon release during hypoglycemia is controlled by glucokinase-mediated glucose sensing outside the brain but not in β-cells. For epinephrine, we found increased responses in GCK-MODY patients, in β-cell ablated diabetic I366F mice and in conditional (nestin lineage) brain glucokinase-knockout mice, supporting a role for brain glucokinase in triggering epinephrine release. Conclusions Our data suggest that glucokinase in brain and other non β-cell peripheral hypoglycemia sensors is important in glucose homeostasis, allowing the body to detect and respond to a falling blood glucose. Graphical abstract Image 1 HIGHLIGHTS • Reduced glucokinase (GCK) function increases hormonal responses to hypoglycemia. • β-cell GCK is responsible for insulin suppression as blood glucose levels fall. • Brain GCK-mediated hypoglycemia-sensing is involved in epinephrine release. • GCK-mediated glucagon secretion involves GCK that is not located in brain or β-cells. [ABSTRACT FROM AUTHOR]

Summary The molecular transducers of benefits from different exercise modalities remain incompletely defined. Here we report that 12 weeks of high-intensity aerobic interval (HIIT), resistance (RT), and combined exercise training enhanced insulin sensitivity and lean mass, but only HIIT and combined training improved aerobic capacity and skeletal muscle mitochondrial respiration. HIIT revealed a more robust increase in gene transcripts than other exercise modalities, particularly in older adults, although little overlap with corresponding individual protein abundance was noted. HIIT reversed many age-related differences in the proteome, particularly of mitochondrial proteins in concert with increased mitochondrial protein synthesis. Both RT and HIIT enhanced proteins involved in translational machinery irrespective of age. Only small changes of methylation of DNA promoter regions were observed. We provide evidence for predominant exercise regulation at the translational level, enhancing translational capacity and proteome abundance to explain phenotypic gains in muscle mitochondrial function and hypertrophy in all ages. [ABSTRACT FROM AUTHOR]

ApoE is a polymorphic protein involved in the metabolism of plasma lipoproteins; the ε4 allele was shown to be associated with coronary and aortic atherosclerosis in age-dependent fashion mediated by unknown mechanisms. This study was undertaken to assess whether the apoE isoforms in humans were associated with normal glucose tolerance and with metabolic and inflammatory risk factors of CVD. ApoE genotype was assessed in 365 individuals. Of those, 309 were studied in the postabsorptive conditions and 142 of them also underwent a 3h-OGTT; 56 additional subjects were studied by means of the insulin clamp in combination with [6,6-2H2] glucose infusion. ApoE genotype frequencies were similar to those previously reported and were not influenced by age and BMI. Fasting plasma glucose, insulin, FFA, the lipid profile, surrogate markers (HOMA-IR, OGTT-derived index) as well as the clamp-derived parameters or insulin sensitivity and insulin secretion were not different by apoE genotypes. Serum adipokines concentrations (leptin, adiponectin, resistin) and markers of inflammation (serum fasting hsCRP and MCP1/CCL2) were also not different by apoE genotypes. In the subgroup of young ε4 carriers which underwent the clamp procedure, a higher fasting endogenous glucose production was detected. ApoE genotype was not associated with insulin resistance or altered insulin secretion, and no abnormalities in the typical circulating endocrine, metabolic, and inflammatory features of the insulin resistance syndrome were detected. [ABSTRACT FROM AUTHOR]

This article addresses two topics. We provide an overview of the National Institutes of Health Mouse Metabolic Phenotyping Center (MMPC) Program. We then discuss some observations we have made during the first eight years of the Vanderbilt MMPC regarding common phenotyping practices. We include specific recommendations to improve phenotyping practices for tests of glucose tolerance and insulin action. We recommend that methods for experiments in vivo be described in manuscripts. We make specific recommendations for data presentation, interpretation, and experimental design for each test. To facilitate and maximize the exchange of scientific information, we suggest that guidelines be developed for methods used to assess glucose tolerance and insulin action in vivo. [ABSTRACT FROM AUTHOR]

Vascular endothelium regulates vascular tone and tissue perfusion in response to various physiological and pathological stimuli. Insulin and meal feeding increase microvascular perfusion and thus oxygen, nutrient, and hormone delivery to human skeletal muscle. Meal feeding also increases cardiac microvascular perfusion in healthy humans. To examine whether insulin at physiological concentrations increases microvascular perfusion in human myocardium, we studied 13 healthy, overnight-fasted, lean, young human volunteers by using myocardial contrast echocardiography (MCE) and insulin-clamp techniques. We measured cardiac microvascular blood volume (MBV), microvascular flow velocity (MFV), and microvascular blood flow (MBF) at baseline, 60 min, and 120 min after initiating insulin infusion at 1 mU∙kg-1∙min-1. MBF is the product of MBV and MFV and represents microvascular perfusion. Insulin increased myocardial MBV by 23% at 60 min (P < 0.01) and by 41% at 120 min (P = 0.001) without changing MFV. As a result, insulin-mediated myocardial MBF increased significantly at both 60 min (P < 0.01) and 120 min (P < 0.0005). Insulin also significantly increased brachial artery diameter, flow velocity, and total blood flow at 60 and 120 min (P < 0.05 for all). The changes in cardiac MBV correlated positively with quantitative insulin sensitivity check index (QUICKI) and negatively with body mass index but not with the steady-state glucose-infusion rates or the changes in brachial artery parameters. We conclude that insulin, at physiologically relevant concentrations, increases microvascular perfusion in human heart muscle by increasing cardiac MBV in healthy, insulin-sensitive adults. This insulin-mediated cardiac microvascular perfusion may play an important role in normal human myocardial oxygen and substrate physiology. [ABSTRACT FROM AUTHOR]

Diet-induced milk fat depression in dairy cows has been known for many years and several theories have been proposed. One that continues to receive support is the glucogenic-insulin theory. Previous studies testing this theory using a hyperinsulinemic--euglycemic clamp have had variable results attributable to variability in the use of body fat reserves as a source of milk fatty acids. Our objective was to test the glucogenic-insulin theory using cows immediately postpartum, a period when the use of body fat for milk fat synthesis is greatest. During wk 2 postpartum, 5 cows were given a 2-d baseline period and then clamped for 4 d. Insulin was increased more than 2-fold during the clamp while the blood glucose concentration was maintained. Milk yield was not altered by administration of the clamp (38.7 vs. 39.0 ± 1.4 kg/d); however, the milk fat percentage and yield were reduced by 27% and plasma nonesterified fatty acids were reduced by 68%. Analysis of the milk fatty acid composition revealed that the decrease in milk fat yield during use of the clamp was almost exclusively due to reductions in preformed fatty acids; this is the exact opposite of what is observed with diet-induced milk fat depression. Therefore, our results do not support the glucogenic-insulin theory of diet-induced milk fat depression. The results further indicated that reductions in milk fat observed previously with hyperinsulinemic-euglycemic clamps or with glucose or propionate infusions were most likely consequences of the ability of insulin to inhibit lipolysis, thereby limiting the mammary availability of preformed fatty acids mobilized from body reserves. [ABSTRACT FROM AUTHOR]

Abstract: This trial was designed to test the effects of insulin, rbST and their interaction on DMI and fat yield and composition in dairy cows. Eight Holstein cows (86±10 DIM) were divided in two groups and used in two replicates of a Latin square design with four animals, four periods and the following four treatments: (1) intravenous infusion of saline, (2) infusion of saline and subcutaneous administration of 40 mg of rbST per day, (3) intravenous infusion of 12 mg of insulin per day coupled with glucose infusion, and (4) rbST administration combined with insulin and glucose infusion. The glucose infusion rate was adjusted to maintain euglycemia. Each experimental period lasted 14 days, i.e., treatments were administered during the first 6 days and no treatment was administered during the following 8-day resting phase. Data from the last 3 days of infusion were analysed using the SAS system for mixed models. For DMI, milk yield, fat percent, lactose percent and lactose yield, there was a significant interaction between insulin and rbST administration. However, fat yield was higher (1.63 vs. 1.39 kg/day), with a higher content of long chain fatty acids (242.2 vs. 176.4 g/milking), during rbST administration, regardless of insulin infusion. Insulin infusion decreased fat yield (1.39 vs. 1.63 kg/day for the control group), mainly through a decrease in long chain fatty acids (179.7 vs. 238.9 g/milking for the control group), regardless of rbST administration. Insulin decreased triglycerides in plasma (0.11±0.01 vs. 0.15±0.01 mmol/L). Plasma non-esterified fatty acids were increased by rbST but this increase was prevented by insulin. Overall, these results clearly indicated that insulin decreased milk fat yield but this decrease, unlike during classical milk fat depression (MFD), is associated only with a decrease in long chain fatty acids in milk. [Copyright &y& Elsevier]

1. Glucose phosphorylation is the first irreversible step of the muscle glucose uptake pathway and is catalysed by a hexokinase isozyme.2. While glucose transport is the primary barrier to muscle glucose uptake during basal conditions, glucose phosphorylation becomes an important barrier to muscle glucose uptake during stimulated conditions such as hyperinsulinaemia or exercise.3. High fat feeding markedly impairs insulin- and exercise-stimulated muscle glucose uptake. As hexokinase II overexpression corrects this dietary-induced deficit during exercise, glucose phosphorylation is a site of impairment following high fat feeding.4. Exercise is an important tool for diagnosing deficits in glucose phosphorylation. [ABSTRACT FROM AUTHOR]

Positron emission tomography (PET) is a unique tool for studying regional skeletal muscle glucose metabolsm and blood flow in vivo. The application of PET in the assessment of skeletal muscle glucose metabolism depends on recent improvements in instrumentation, data analysis, and production of 18F-fluorodeoxyglucose (FDG) and 15O water. The data presented support the validity of the 15O water model to measure blood flow and the FDG model for the determination of glucose uptake and glucose kinetic constants (influx, efflux, and phosphorylation) in skeletal muscle. However, quantification of absolute glucose transport and backflux rates should be applied with caution because those calculations are based on unproven assumptions such as validity of the lumped constant for these individual processes and constancy of the free and accessible intracellular glucose pool. It is evident that quantification of glucose fluxes using the triple tracer technology generates conflicting data that violate assumptions inherent in triple tracer or PET modeling. Further FDG-PET studies will have to solve those problems to provide more insight into the regulatory processes of glucose transport and phosphorylation of different insulin-resistant disease states. Promising new areas of PET research will include not only detailed study of glucose kinetics but also the measurement of muscle protein synthesis in vivo, which is of interest in a variety of conditions. [ABSTRACT FROM AUTHOR]

Aims/hypothesis: We aimed to examine whether impaired glucose tolerance is associated with reduced suppression of glucagon concentrations. Methods: Eighty-four non-diabetic women of Caucasian origin and 61 years of age, of whom 48 had normal glucose tolerance (NGT) and 36 had IGT, underwent a 75 g OGTT and a hyperinsulinaemic, euglycaemic clamp with measurement of glucagon, insulin and glucose concentrations. Results: At 2 h after 75 g oral glucose, glucagon concentrations were reduced by 7.1 ± 1.1 ng/l in NGT vs 8.0 ± 1.4 ng/l in IGT, (NS). However, the 2 h reductions in glucagon per mmol/l increase in 2 h glucose or per pmol/l increase in 2 h insulin were both impaired in IGT (p = 0.002 and p = 0.043, respectively) because the 2 h increases in glucose and insulin were higher in IGT than in NGT. Furthermore, suppression of glucagon concentrations during a euglycaemic clamp at hyperinsulinaemic concentrations (NGT: 607 ± 19 pmol/l, IGT: 561 ± 21 pmol/l) was lower in IGT (13.6 ± 1.6 ng/l) than in NGT (23.1 ± 1.2 ng/l; p < 0.001). The suppression of glucagon concentrations during the hyperinsulinaemic, euglycaemic clamp correlated with insulin sensitivity (r = 0.24, p = 0.027) and with the 2 h glucose value during the OGTT (r = –0.52, p < 0.001). Conclusion/interpretation: Impaired glucose tolerance is associated with reduced insulin-induced suppression of glucagon secretion, which could be caused by A-cell insulin resistance. Inappropriately high glucagon secretion could therefore contribute to the metabolic perturbations in IGT. [Diabetologia (2001) 44: 1998–2003] [ABSTRACT FROM AUTHOR]

Increased deposition of the extracellular matrix (ECM) in adipose tissue (AT) during obesity contributes to insulin resistance. The integrin receptors transmit changes in the extracellular environment causing corresponding intracellular adaptations. Integrin-linked kinase (ILK), an adaptor protein, is a central hub for intracellular signaling of integrins. This study determined the role of ILK in adipose function and insulin resistance. The pathogenic role of ILK in obesity and insulin resistance was studied in human adipose tissue and adipocyte-specific ILK-deficient mice (ILKlox/lox AdCre). ILKlox/lox AdCre mice together with wild-type littermates (ILKlox/lox) were fed a chow diet or 60% high-fat (HF) diet for 16 weeks. In vivo insulin sensitivity was determined by hyperinsulinemic-euglycemic clamps. AT ILK expression was increased by HF diet feeding in mice and increased in visceral fat of morbidly obese humans. The HF-fed ILKlox/lox AdCre mice displayed reduced fat mass and improved glucose tolerance relative to the HF-fed ILKlox/lox mice. During a hyperinsulinemic-euglycemic clamp, the HF-fed ILKlox/lox AdCre mice exhibited partially improved insulin resistance in AT. Lipolysis was suppressed to a greater extent by insulin and glucose uptake in brown AT (BAT) increased. Increased inhibition of lipolysis may have been attributed to increased vascularization in white AT, while increased glucose uptake in BAT was associated with increased Akt phosphorylation and P38/JNK dephosphorylation. Notably, AT insulin sensitivity in lean mice was not affected by ILK deletion. Moreover, reduced fat mass in the HF-fed ILKlox/lox AdCre mice may have been attributed to decreased free fatty acid uptake into adipocytes via the downregulation of CD36 gene expression. Consistent with the results in the mice, knockdown and knockout of ILK in 3T3-L1 cells decreased lipid accumulation and CD36 gene expression during adipogenesis. These data show that adipocyte ILK is important for regulating HF diet-mediated insulin resistance in AT in a manner consistent with AT function. • ILK protein increased in visceral adipose tissue of obese humans and mice. • Mice lacking adipocyte ILK had less fat and improved glucose tolerance in obesity. • Adipocyte ILK deletion improved anti-lipolytic action of insulin in obese mice. • Adipocyte ILK deletion stimulated brown adipose tissue glucose uptake in obese mice. [ABSTRACT FROM AUTHOR]

To evaluate the role of abnormal glucagon physiology in the glucose intolerance of aging, we have examined: (1) basal glucagon concentration, (2) suppression of plasma glucagon by hyperglycaemia and hyperinsulinaemia, (3) plasma glucagon response to intravenous alanine, (4) glucagon kinetics, and (5) hepatic sensitivity to glucagon (3 ng·kg·min for 3 h) using 3-H-glucose in young (24±1 years), middleaged (41±2 years), and older (62±2 years) subjects. Fasting plasma glucagon levels in 58 young, 21 middle-aged, and 32 older subjects were 89±10, 91±12, and 96±21 pg/ml, respectively ( p=NS). Following elevation of plasma glucose to 2.2-6.9 mmol/l above basal for 2 h (hyperglycaemic clamp technique), young, middle-aged, and older subjects showed similar depressions in plasma glucagon levels during all three studies. Elevation of the plasma insulin concentration to approximately 100 mU/l while maintaining euglycaemia (insulin clamp technique) also resulted in a similar decline in plasma glucagon levels in both young and older subjects (by 25±3 and 27±3 pg/ml respectively). Following intravenous alanine infusion, the increase in plasma glucagon was not significantly different in young compared with older subjects (65±16 versus 38±7 pg/ml). Plasma insulin (7±1 versus 8±1 mU/l) and glucose (0.33±0.11 versus 0.50±0.11 mmol/l) responses were comparable in young and older subjects. During continuous glucagon infusion, the steady-state plasma glucagon concentration (267±32 versus 252±12 pg/ml) and the metabolic clearance rate of glucagon (16±1 versus 15±1 ml· kg·min) were similar in young and older subjects. However, the increase in plasma glucose concentration was significantly greater in the older than young subjects (Δ=1.17±0.11 versus 0.67±0.06 mmol/l, p<0.01). Basal glucose production in older subjects (0.13 ±0.01 mmol·kg·min) was slightly less than in the young (0.15±0.01 mmol·kg· min). However, the rise in glucose production following glucagon was significantly greater in older than young subjects (31±5 versus 18±4%, p<0.05). In conclusion, (1) basal glucagon levels are not consistently elevated in older subjects, (2) glucagon suppression by both hyperglycaemia and hyperinsulinaemia is similar in young and older subjects, (3) α cell sensitivity to alanine is not affected by age, (4) the metabolic clearance rate of glucagon is similar in young and older subjects, and (5) hepatic sensitivity to a physiological increment in plasma glucagon is increased in older subjects. [ABSTRACT FROM AUTHOR]

In the arginine-stimulated perfused rat pancreas, elevated concentrations of potassium ion inhibited glucagon secretion while stimulating the secretion of insulin. Decreased potassium ion produced the reverse effect. The observed inverse correlation between changes in insulin and glucagon secretion ( r = -0.64; p<0.001) was suggestive of local interactions between islet hormones, and prompted us to determine whether potassium-induced changes in glucagon secretion were dependent upon concurrent changes in insulin release. We found that when insulin secretion was greatly suppressed, either through acute induction of diabetes with streptozotocin or by utilization of a glucose-free perfusate, no qualitative differences in glucagon responsiveness to altered potassium ion were evident, although the amplitude of these glucagon changes was enhanced. Similarly, when exogenous insulin (20,000 mU/l) was added to the perfusate in order to render potassium-induced changes in endogenous insulin secretion insignificant, glucagon responsiveness to altered potassium ion was maintained. Exogenous insulin alone had no effect on arginine-stimulated glucagon secretion. We conclude that any indirect effects of potassium ion on arginine-stimulated glucagon secretion are not mediated by insulin, but could be related to changes in somatostatin secretion. [ABSTRACT FROM AUTHOR]

We studied the effects of prior high-intensity cycle exercise (85% VO2 max) on total energy expenditure (EE) and the thermic effect of insulin (TEl) in normal controls and in obese insulin-resistant and noninsulin-dependent diabetic (NIDDM) subjects. Normal controls, but not obese or NIDDM subjects, showed a significant increase (3-7%) in total EE 12-16 h after exercise (p <0.05). Prior exercise increased lipid and decreased glucose oxidation in all groups. Daring low-dose insulin infusion (40 mU • M2 • mm), prior exercise potentiated TEl in control and NIDDM subjects (p <0.05), whereas obese subjects showed no response. During high-dose insulin infusion (40 mU • M² • mm), TEl was similar in NIDDM and control subjects but was significantly less in the obese group. In this study, we found a positive correlation between TEl and insulin-stimulated rates of glucose disposal (r = 0.91, p <0.001). The predicted cost of glucose storage accounted for 42% of TEI. [ABSTRACT FROM AUTHOR]

We propose a new quantitative method for L-lactate assay in whole blood, based on the measurement of pH variation caused by specific and irreversible oxidation of L-lactate to pyruvate in the presence of an electron acceptor (hexacyanoferrate) and of the enzyme cytochrome b2 (EC 1.1.2.3.). No sample pretreatment is needed; the method is simple and fast (1.5 min/analysis) and requires 10 µl whole blood per assay. Linearity is confirmed up to 20 mmol/l L-lactate. Within-day and between-day variability was (as C.V.) 3.6% and 8.1% for blood lactate 1.3 and 1.0 mmol/l, respectively. The results by the present method correlate well with those from two reference methods (test method vs a lactate sensor based method: r=0.996; test method vs a spectrophotometric method: r=0.987). An application of the present method to the continuous monitoring of L-lactate in patients after combined kidney and pancreas transplantation, under conditions of euglycemic hyperinsulinemia and hyperglycemic hyperinsulinemia is reported. We conclude that the method is simple and reproducible and can be employed to measure whole blood lactate concentration continuously both in clinical protocols and in basic research. [ABSTRACT FROM AUTHOR]

In a previous study, we observed an impairment of the theophylline-induced suppressive system in recent onset IDDM patients, and demonstrated also a correlation with metabolic derangement. The aim of this study was to better investigate the relationship between theophylline sensitivity (ThS) and blood glucose/plasma insulin levels in recent onset IDDM patients subjected to preprogrammed variations by an insulin/glucose clamp with artificial pancreas. Eight patients were studied within 8 weeks from the onset of IDDM. ThS was evaluated as the ability of theophylline to inhibit blastogenic response of peripheral blood lymphocytes (PBL) to Concanavalin A (ConA), after 120 min preincubation of the cells. All patients were connected to an artificial pancreas. Through i.v. continuous insulin infusion (0.02 U/kg/h) and/or i.v. continuous glucose and saline infusion, the following experimental conditions, lasting at least 1h, were obtained: T1: relative euglycemia and normal insulinemia; T2: relative euglycemia and hyperinsulinemia; T3: hyperglycemia and normal insulinemia; T4: hyperglycemia and hyperinsulinemia. ThS was maintained in 6/8 patients at T1 and in 8/8 patients at T4. ThS was lost in 4/8 patients at T2 and T3. These data suggest that the loss of ThS induced by hyperglycemia can be corrected by hyperinsulinemia, and that it is maintained when euglycemia is accompained by hypoinsulinemia. It is lost when these two parameters lose their interrelationship. [ABSTRACT FROM AUTHOR]

Introduction: Hepatic insulin clearance (HIC) is an important pathophysiology of type 2 diabetes. HIC was reported to decrease in patients with type 2 diabetes and metabolic syndrome. However, hyperglycemia was suggested to enhance HIC, and it is not known whether poorly controlled diabetes increases HIC in patients with type 2 diabetes. We investigated whether HIC was increased in patients with poorly controlled diabetes, and whether HIC was associated with insulin resistance and incretins., Research Design and Methods: We performed a meal tolerance test and the hyperinsulinemic-euglycemic clamp in 21 patients with type 2 diabetes. We calculated the postprandial C-peptide area under the curve (AUC)-to-insulin AUC ratio as the HIC; measured fasting and postprandial glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) and glucagon levels and analyzed serum adiponectin and zinc transporter-8 (ZnT8) gene polymorphism., Results: The HIC significantly correlated with glycated hemoglobin (HbA1c) ( r&#95;S =0.58, p<0.01). In patients with high HIC above the median of 6.5, the mean HbA1c was significantly higher compared with low HIC below the median. Homeostatic model assessment (HOMA)-beta ( r&#95;S =-0.77, p<0.01) and HOMA-IR ( r&#95;S =-0.66, p<0.005) were correlated with HIC. The M/I value in the clamp study was correlated with HIC. GLP-1-AUC and GIP-AUC were not correlated with HIC. Glucagon-AUC was negatively correlated with HIC, but there were no significant differences between the high and low HIC groups. Adiponectin was positively correlated with HIC. The ZnT8 gene polymorphism did not affect HIC., Conclusions: These results suggest that HIC was increased in patients with high HbA1c type 2 diabetes, low insulin secretion, low insulin resistance and high adiponectin conditions., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)