Glycogen content and activities of key glycolytic enzymes in muscle biopsies from control subjects and patients with chronic alcoholic skeletal myopathy

The capacity for glycolysis in muscle biopsies obtained from long-term heavy alcohol drinking patients has been compared with tissue from control subjects by assay in vitro of the total activities of glycogen phosphorylase, phosphofructokinase and fructose 1,6-bisphosphatase, key regulatory enzymes in the anaerobic glycolytic pathway. Biopsies from 13 of 22 patients had type II fibre atrophy, and the activities of all three enzymes were reduced in these biopsies, when expressed in terms of DNA content, the most striking reduction being in phosphofructokinase activity. The amount of glycogen in the tissue correlated closely with these enzyme activities and was slightly lower in the most atrophic tissue, when expressed in terms of DNA content. The activities of acid and neutral alpha-glucosidases were similar in biopsies from control subjects and patients with various severities of alcohol myopathy. The reduced activities are consistent with a reduced proportion of type II fibre muscle mass in these patients, and suggest that there may be a reduced capacity for glycolysis with resultant reduced lactate production. Whether the changes in enzyme activities are primary to the selective atrophy remains to be established.     

Metabolic changes in human liver associated with preoperative intravenous nutrition

1. This study examined the influence of pre-operative intravenous nutrition upon carbohydrate stores, glucose metabolism and protein synthesis in the liver of patients undergoing laparotomy. 2. Thirty patients with gastrointestinal cancer and weight loss (greater than 5 kg in 3 months) were randomized to receive a hospital diet only or a hospital diet plus intravenous nutrition (0.18 g of N + 125 kJ day-1 kg-1) for 3 or 7 days before laparotomy. Patients who had not lost weight received the hospital diet only and formed a control group. 3. Wedge biopsies of liver were obtained at laparotomy and analysed for glycogen concentration, the activity of three key enzymes of glucose metabolism, 6-phosphofructokinase (EC 2.7.1.11), fructose bisphosphatase (EC 3.1.3.11) and hexokinase (EC 2.7.1.1), and the capacity for protein synthesis. 4. Compared with controls and the hospital diet group, both phosphofructokinase and fructose bisphosphatase activity were reduced in patients who received intravenous nutrition, suggesting the utilization of glucose for glycogen synthesis with a reduction in the glycolytic flux. Consistent with these changes, patients who received intravenous nutrition had a significantly higher glycogen concentration compared with the control and hospital diet groups. 5. Maximal rates of protein synthesis were achieved after only 3 days of intravenous nutrition. 6. The provision of intravenous nutrition was associated with changes in hepatic metabolism suggestive of repletion of energy stores and a higher capacity for protein synthesis.     

Phosphofructokinase deficiency associated with congenital nonspherocytic hemolytic anemia and mild myopathy: biochemical and morphological studies on the muscle

Enzymatic and electron microscopical studies were performed on the muscle of a proband with phosphofructokinase deficiency. Enzymatic studies showed that muscle phosphofructokinase activity of the proband was decreased to about a half of normal. This enzyme was quite thermolabile and had low affinity for fructose 6-phosphate. Electron microscopical studies showed the accumulation of glycogen granules beneath the sarcolemma and between the myofibrils in spite of a lack of accumulation of the intermediates before the step of phosphofructokinase. The proband's clinical symptoms, i.e., hemolytic anemia and myopathy, were considered to be due to the unstable, mutant, muscle-type phosphofructokinase in the red blood cells and muscle.     

Study of skeletal muscle glycogenolysis and glycolysis in chronic steroid myopathy, non-steroid histochemical type-2 fiber atrophy, and denervation

OBJECTIVE: Muscle biopsies from chronic steroid (glucocorticoid) myopathy, non-steroid histochemical type-2 fiber atrophy, and muscle denervation patients were studied to determine if their glycogen contents, or enzymes involved in glycogenolysis and glycolysis might be related to their fiber atrophy. DESIGN AND METHODS: Fast frozen muscle biopsies from the above patients and from patients later judged by histochemistry to be normal were assayed enzymatically for glycogen content, for enzymes involved in glycogenolysis, and for 6 of the enzymes involved in glycolysis. RESULTS AND CONCLUSION: All three groups of patients had glycogen content, but only the chronic steroid myopathy muscle had statistically less glycogen content than did normal human muscle. All 3 groups had statistically low mean values compared to normal muscles for glycogen phosphorylase activity. This suggests that the biosynthesis and phosphorolysis of glycogen are not involved in muscle fiber atrophy, and glucocorticoid administration does not activate muscle glycogen biosynthesis. Histochemical type-2 fiber atrophy muscles were low compared to normal muscles in three glycogenolysis enzyme activities plus four glycolysis enzyme activities. Muscles from denervation patients were low compared to normal muscles in three glycogenolysis enzyme activities plus five glycolysis enzyme activities. This suggests that muscle denervation may lower the rate of glycolysis enough to fail to provide sufficient pyruvate for mitochondrial ATP biosynthesis, resulting in insufficient protein biosynthesis in both fiber types.     

Glycogen synthase and phosphofructokinase protein and mRNA levels in skeletal muscle from insulin-resistant patients with non-insulin-dependent diabetes mellitus.

In patients with non-insulin-dependent diabetes mellitus (NIDDM) and matched control subjects we examined the interrelationships between in vivo nonoxidative glucose metabolism and glucose oxidation and the muscle activities, as well as the immunoreactive protein and mRNA levels of the rate-limiting enzymes in glycogen synthesis and glycolysis, glycogen synthase (GS) and phosphofructokinase (PFK), respectively. Analysis of biopsies of quadriceps muscle from 19 NIDDM patients and 19 control subjects showed in the basal state a 30% decrease (P < 0.005) in total GS activity and a 38% decrease (P < 0.001) in GS mRNA/microgram DNA in NIDDM patients, whereas the GS protein level was normal. The enzymatic activity and protein and mRNA levels of PFK were all normal in diabetic patients. In subgroups of NIDDM patients and control subjects an insulin-glucose clamp in combination with indirect calorimetry was performed. The rate of insulin-stimulated nonoxidative glucose metabolism was decreased by 47% (P < 0.005) in NIDDM patients, whereas the glucose oxidation rate was normal. The PFK activity, protein level, and mRNA/microgram DNA remained unchanged. The relative activation of GS by glucose-6-phosphate was 33% lower (P < 0.02), whereas GS mRNA/micrograms DNA was 37% lower (P < 0.05) in the diabetic patients after 4 h of hyperinsulinemia. Total GS immunoreactive mass remained normal. In conclusion, qualitative but not quantitative posttranslational abnormalities of the GS protein in muscle determine the reduced insulin-stimulated nonoxidative glucose metabolism in NIDDM.     

Glucose tolerance in relation to skeletal muscle enzyme activities in cancer patients.

Glucose metabolism and skeletal muscle enzyme activities were studied in nineteen cancer patients and twelve matched controls. The fasting insulin values were normal but the fasting glucose values and the sum of glucose were increased and the sum of insulin was decreased during intravenous glucose tolerance test in the cancer patients. The elimination rate of glucose (k-value) during glucose challenge was, however, not significantly different in cancer patients as compared with that of appropriate controls. The activities of enzymes representative for glycogen turnover, glycolysis, citric acid cycle and respiratory chain were significantly lower in the muscle tissue of cancer patients, while the activity of 3-hydroxyacyl-CoA dehydrogenase, an enzyme in the beta-oxidation of fatty acids, was unchanged and the activity of glucose-6-phosphate dehydrogenase was significantly higher. Rate limiting enzyme activities in muscle tissue, phosphofructokinase and cytochrome c oxidase correlated signficantly with plasma insulin and glucose during glucose challenge. The results point at the possibility of covariating debilitation of pancreatic beta-cells and skeletal muscle enzymes caused by the malignant tumour.     

Glycogen and lactate synthetic pathways in human skeletal muscle in relation to obesity, weight reduction and physical training

The effects of obesity, weight reduction, and physical condition on the concentrations of glucose-6-phosphate (G-6-P) and glycogen, and the activities of glycogen synthase (GS) and lactate dehydrogenase (LD) were determined in resting vastus or gastrocnemius muscles of 40 healthy subjects. In obese women the activity of GS was 50% (P less than 0.05) lower than in lean women with similar levels of glycogen and G-6-P, whereas no difference was found in the activity of LD. Calorie restriction induced a 4.5% (P less than 0.05) decrease in body weight from 82.5 kg corresponding to a 3.2% (P less than 0.05) decrease in body mass index from 30.9 kg m-2. The total and fractional activities of glycogen synthase were increased by 50% (P less than 0.05), whereas muscle glycogen content was reduced by 40% (P less than 0.05). The G-6-P concentration and the activity of LD remained unchanged. In well-trained young men the concentrations of G-6-P and glycogen were, respectively, 250% (P less than 0.05) and 50% (P less than 0.05) higher than in non-trained. The fractional and total activities of GS were 90% (P less than 0.05) and 50% (P less than 0.05) higher, respectively, and the total activity of LD was only half (P less than 0.05) that of non-trained subjects. In conclusion, physical training enhances the activity of GS, despite a concomitantly increased glycogen content, and thus seems to exert a more efficient stimulus on glycogen synthase than weight reduction. It is indicated that physical training may provide a clinically important contribution to blood glucose reduction in hyperglycaemic conditions.     

Defective insulin response of phosphorylase phosphatase in insulin-resistant humans.

Insulin-stimulated glycogen synthase activity in human muscle is reduced in insulin-resistant subjects. Insulin regulation of human muscle glycogen synthase may require activation of a type-1 protein phosphatase (PP-1). We investigated the change of phosphorylase phosphatase and glycogen synthase activities in muscle biopsies obtained during a 2-h hyperinsulinemic euglycemic clamp in 12 insulin-sensitive (group S) and 8 insulin-resistant (group R) subjects. Fasting phosphorylase phosphatase activity was lower in group R than in group S, and did not increase significantly with insulin infusion in group R until 20 min. In group S, phosphorylase phosphatase was significantly stimulated by 10 min, remaining significantly higher than in group R at all time points. The insulin-mediated changes in phosphatase activities were not decreased by 3 nM okadaic acid but were completely inhibited by 1 microM okadaic acid, thereby verifying that insulin-stimulated phosphorylase phosphatase is accounted for by a PP-1. Subcellular fractionation demonstrated reduced fasting PP-1 activities in both the glycogen and cytosolic fractions of muscle obtained from subjects in group R compared to those in group S. These results suggest that insulin activation of PP-1 could contribute to the stimulation of glycogen synthase by this hormone in human muscle. Lower fasting PP-1 activity in cytosol and glycogen fractions plus lower insulin-stimulated PP-1 activity could explain, in part, reduced insulin-stimulated glycogen synthase in skeletal muscle of insulin-resistant subjects.     

Quercitrin,a bioflavonoid improves glucose homeostasis in streptozotocin‐induced diabetic tissues by altering glycolytic and gluconeogenic enzymes

The present study is an investigation into the role of quercitrin on carbohydrate metabolism in normal and streptozotocin (STZ)‐induced diabetic rats. Administration of STZ leads to a significant increase (P < 0.05) in fasting plasma glucose and a decrease in insulin levels. The content of glycogen is significantly decreased (P < 0.05) in liver and muscle, but increased in the kidney. The activity of hexokinase decreased whereas the activities of glucose 6‐phosphatase and fructose 1,6‐bisphosphatase significantly increased (P < 0.05) in the tissues. Oral administration of quercitrin (30 mg/kg) to diabetic rats for a period of 30 days resulted in significant (P < 0.05) alterations in the parameters studied but not in normal rats. A decrease of plasma glucose and increase in insulin levels were observed along with the restoration of glycogen content and the activities of carbohydrate metabolic enzymes in quercitrin‐treated diabetic rats. The histopathological study of the pancreas revealed the protective role of quercitrin. There was an expansion of the islets and decreased fatty infiltrate of the islets in quercitrin treated diabetic rats. In normal rats treated with quercitrin, we could not observe any significant change in all the parameters studied. Combined, these results show that quercitrin plays a positive role in carbohydrate metabolism and antioxidant status in diabetic rats.     

Effect of hyperthyroidism on fibre-type composition,fibre area,glycogen content and enzyme activity in human skeletal muscle

Summary. Seven hyperthyroid patients were studied by repeated muscle biopsies (vastus lateralis) before and after a period of medical treatment which averaged 10 months. The biopsies were analysed with regard to fibre-type composition, fibre area, capillary density, glycogen content and enzyme activities representing the glycolytic capacity (hexokinase, 6-phosphofructokinase), oxidative capacity (oxoglutarate dehydrogenase, citrate synthase) and Ca²⁺- and Mg²⁺-stimulated ATPase in muscle. In the pretreatment biopsy (hyperthyroid state), there was a significantly lower proportion of type I fibres (30% vs. 41%), a higher capillary density (23%), lower glycogen content (33%), and higher hexokinase activity (32%) compared with the post-treatment biopsy. No significant changes in the activity of the remaining enzymes were observed. The present study indicates that hyperthyroidism induces a transformation from type I to type II fibres in human skeletal muscle. The increase in hexokinase activity probably reflects a higher glucose utilization by skeletal muscle in order to compensate partially for the reduced glycogen content.     

Specificity of recumbent cycling as a training modality for the functional movements; sit-to-stand and step-up

BACKGROUND: The principle of specificity in muscle training requires the training mode to reflect the desired outcome. The observed similarity of lower limb movements during recumbent cycling to the functional movements sit-to-stand and step-up presents the possibility of using recumbent cycling in a rehabilitation context. This may reduce the need to practice the actual task which in some, less able, patients may be labour intensive and patient fatiguing. To date no studies have compared recumbent cycling to these functional movements. This study therefore aimed to compare the lower limb kinematics and muscle activity between recumbent cycling and both sit-to-stand and step-up movements. METHODS: Electromyographic and kinematic signals from 12 young (mean age 42.1 years) healthy participants were collected during the performance of three activities: (1) cycling at 60 rpm, (2) sit-to-stand and (3) a single step-up. Only the extension phase of each movement was compared. FINDINGS: Although the results demonstrated differences in joint movement and muscle activation, e.g., greater gastrocnemius activity during recumbent cycling (P<0.00), knee range of motion and average root mean square activity for rectus femoris, biceps femoris and the sum of the average activity for five muscles recorded showed no difference (P>0.05) suggesting that there was sufficient agreement to support the use of recumbent cycling as a specific training modality for the sit-to-stand and step-up movements. This finding may have positive implications for the rehabilitation of a wide range of patients in the early stages of rehabilitation.     

Mechanism of impaired insulin-stimulated muscle glucose metabolism in subjects with insulin-dependent diabetes mellitus.

To determine the mechanism of impaired insulin-stimulated muscle glycogen metabolism in patients with poorly controlled insulin-dependent diabetes mellitus (IDDM), we used 13C-NMR spectroscopy to monitor the peak intensity of the C1 resonance of the glucosyl units in muscle glycogen during a 6-h hyperglycemic-hyperinsulinemic clamp using [1-(13)C]glucose-enriched infusate followed by nonenriched glucose. Under similar steady state (t = 3-6 h) plasma glucose (approximately 9.0 mM) and insulin concentrations (approximately 400 pM), nonoxidative glucose metabolism was significantly less in the IDDM subjects compared with age-weight-matched control subjects (37+/-6 vs. 73+/-11 micromol/kg of body wt per minute, P < 0.05), which could be attributed to an approximately 45% reduction in the net rate of muscle glycogen synthesis in the IDDM subjects compared with the control subjects (108+/-16 vs. 195+/-6 micromol/liter of muscle per minute, P < 0.001). Muscle glycogen turnover in the IDDM subjects was significantly less than that of the controls (16+/-4 vs. 33+/-5%, P < 0.05), indicating that a marked reduction in flux through glycogen synthase was responsible for the reduced rate of net glycogen synthesis in the IDDM subjects. 31P-NMR spectroscopy was used to determine the intramuscular concentration of glucose-6-phosphate (G-6-P) under the same hyperglycemic-hyperinsulinemic conditions. Basal G-6-P concentration was similar between the two groups (approximately 0.10 mmol/kg of muscle) but the increment in G-6-P concentration in response to the glucose-insulin infusion was approximately 50% less in the IDDM subjects compared with the control subjects (0.07+/-0.02 vs. 0.13+/-0.02 mmol/kg of muscle, P < 0.05). When nonoxidative glucose metabolic rates in the control subjects were matched to the IDDM subjects, the increment in the G-6-P concentration (0.06+/-0.02 mmol/kg of muscle) was no different than that in the IDDM subjects. Together, these data indicate that defective glucose transport/phosphorylation is the major factor responsible for the lower rate of muscle glycogen synthesis in the poorly controlled insulin-dependent diabetic subjects.     

Muscle citrate content and the regulation of metabolism in fed and fasted human skeletal muscle

Summary. Blood and muscle metabolite levels were measured in seven healthy young adult male subjects in a fed state and again following a 24-hour fast. Skeletal muscle samples were obtained under local anaesthesia from m. vastus lateralis using a needle biopsy technique. The blood glucose concentration fell during fasting; the blood lactate concentration remained unchanged. Plasma free fatty acid concentrations rose, as did 3-hydroxybutyrate and acetoacetate levels. There was no change in the amount of carbohydrate stored as glycogen in the muscle in response to fasting. The muscle content of phos-phagens (adenosine triphosphate and phosphocreatine), glycolytic intermediates (glucose-6-phosphate, fructose-6-phosphate, fructose-1, 6-diphosphate, triose phosphates and lactate) and citrate was also not affected by fasting. There was a significant increase in the muscle content of 3-hydroxybutyrate. These results give no indication as to the mechanism by which a decreased rate of carbohydrate degradation might occur in muscle in the fasted state. It is clear, however, that an intracellular accumulation of citrate and a consequent inhibition of glycolysis at the level of phosphofructokinase does not take place in fasting human skeletal muscle.     

Comparative Study of the Metabolism of U-14C-Fructose, U-14C-Sorbitol and U-14C-Xylitol in the Normal and in the Streptozotocin-Diabetic Rat*

Abstract. Young male rats metabolize xylitol, sorbitol and fructose with a half-life of 165 seconds. Within 10 min. after the intravenous injection of a load of these substrates more than 70% of the carbon-14 in plasma is present as ¹⁴C-glucose. The labelling of plasma glucose from these substrates appears to be insulin-independent. A small percentage of labelled xylitol, sorbitol and fructose is retained by the liver as glycogen and total lipids. In the normal rat, this process appears to be insulin-independent. In the chronic streptozotocin-diabetic rat, insulin acutely enhances glycogen synthesis from all three substrates and reduces their incorporation into total lipids. Once these substrates have been converted to glucose the further storage as muscle glycogen and adipose tissue total lipids is entirely dependent on insulin, both in the normal and in the streptozotocin-diabetic rat. Fructose makes an exception since it is metabolized by adipose tissue independently of insulin. In the light of these results in the rat a re-evaluation of the use of various carbohydrates and polyols in parenteral nutrition of normal and diabetic subjects appears desirable.     

The polymorphonuclear leukocyte in diabetes mellitus

In polymorphonuclear leukocytes from severely diabetic patients the rate of glycolysis is decreased due to decreased activity of phosphofructokinase, and the glycogen content and rate of glycogen synthesis are decreased due to a decreased total activity of glycogen synthase and an impaired activation of this enzyme. Covalent modification of glycogen synthase by phosphorylation creates a continuum of phosphorylated enzyme forms of decreasing activity. Phosphorylation of a single peptide, whether by the synthase kinase or the cyclic AMP dependent protein kinase, is critical for the associated kinetic changes during the initial phosphorylation. Conversely, dephosphorylation of this particular peptide is associated with complete activation. The protein phosphatase activity of the microsomal fraction may be separated into functionally and possibly also structurally different phosphorylase- and synthase-phosphatase activities, where the latter appears to be dependent on free cytoplasmic Ca2+. It is hypothesized that it is synthase-phosphatase activity that is absent in leukocytes from diabetic patients and is restored upon insulin treatment.     

Effect of the antilipolytic nicotinic acid analogue acipimox on whole-body and skeletal muscle glucose metabolism in patients with non-insulin-dependent diabetes mellitus.

Increased nonesterified fatty acid (NEFA) levels may be important in causing insulin resistance in skeletal muscles in patients with non-insulin-dependent diabetes mellitus (NIDDM). The acute effect of the antilipolytic nicotinic acid analogue Acipimox (2 X 250 mg) on basal and insulin-stimulated (3 h, 40 mU/m2 per min) glucose metabolism was therefore studied in 12 patients with NIDDM. Whole-body glucose metabolism was assessed using [3-3H]glucose and indirect calorimetry. Biopsies were taken from the vastus lateralis muscle during basal and insulin-stimulated steady-state periods. Acipimox reduced NEFA in the basal state and during insulin stimulation. Lipid oxidation was inhibited by Acipimox in all patients in the basal state (20 +/- 2 vs. 33 +/- 3 mg/m2 per min, P less than 0.01) and during insulin infusion (8 +/- 2 vs. 17 +/- 2 mg/m2 per min, P less than 0.01). Acipimox increased the insulin-stimulated glucose disposal rate (369 +/- 49 vs. 262 +/- 31 mg/m2 per min, P less than 0.01), whereas the glucose disposal rate was unaffected by Acipimox in the basal state. Acipimox increased glucose oxidation in the basal state (76 +/- 4 vs. 50 +/- 4 mg/m2 per min, P less than 0.01). During insulin infusion Acipimox increased both glucose oxidation (121 +/- 7 vs. 95 +/- 4 mg/m2 per min, P less than 0.01) and nonoxidative glucose disposal (248 +/- 47 vs. 167 +/- 29 mg/m2 per min, P less than 0.01). Acipimox enhanced basal and insulin-stimulated muscle fractional glycogen synthase activities (32 +/- 2 vs. 25 +/- 3%, P less than 0.05, and 50 +/- 5 vs. 41 +/- 4%, P less than 0.05). Activities of muscle pyruvate dehydrogenase and phosphofructokinase were unaffected by Acipimox. In conclusion, Acipimox acutely improved insulin action in patients with NIDDM by increasing both glucose oxidation and nonoxidative glucose disposal. This supports the hypothesis that elevated NEFA concentrations may be important for the insulin resistance in NIDDM. The mechanism responsible for the increased insulin-stimulated nonoxidative glucose disposal may be a stimulatory effect of Acipimox on glycogen synthase activity in skeletal muscles.     

Acceptability,safety, and feasibility of in-bed cycling with critically ill patients

BackgroundIn-bed cycling is a promising intervention that may assist critically ill patients to maintain muscle mass and improve their trajectory of recovery. The acceptability of in-bed cycling from the different perspectives of patients, clinicians, and families are unknown. In addition, the safety and feasibility of in-bed cycling in an Australian tertiary intensive care unit (ICU) is relatively unknown.ObjectivesThe objective of this study was to examine the acceptability, safety, and feasibility of in-bed cycling in an Australian tertiary, adult, mixed medical, surgical, trauma ICU.MethodsAn observational process evaluation was embedded in one arm of a two-arm parallel phase II randomised controlled trial that was conducted in an Australian tertiary ICU. The process evaluation was of the acceptability, safety, and feasibility of passive and active in-bed cycling for participants allocated to the trial intervention group. In-bed cycling acceptability questionnaires were designed through a three-step Delphi process. Questionnaire responses from patients, family members, and clinicians who participated in or observed the intervention during the Critical Care Cycling Study (CYCLIST) were evaluated to determine the acceptability of in-bed cycling. The congruence of responses between respondents was also compared. Safety and feasibility of the in-bed cycling intervention were assessed against predetermined criteria.ResultsAcceptability questionnaire responses demonstrated that in-bed cycling was an acceptable intervention from the perspectives of patients, family members, and clinicians. Questionnaire responses were congruent across the respondent groups. Safety was demonstrated with two minor transient adverse events occurring during 276 in-bed cycling sessions (adverse event rate: 0.7%). In-bed cycling sessions were feasible with 276 of 304 (90%) planned sessions conducted.ConclusionsAcceptability questionnaire responses found that in-bed cycling was regarded as an acceptable intervention to patients, family members, and clinicians. The implementation of in-bed cycling was safe and feasible to complete with critically ill patients during the early stages of their critical illness in an Australian tertiary ICU setting.     

Defective insulin response of cyclic adenosine monophosphate-dependent protein kinase in insulin-resistant humans.

Insulin-stimulated glycogen synthase activity in human muscle correlates with insulin-mediated glucose disposal and is reduced in insulin-resistant subjects. Inhibition of the cyclic AMP-dependent protein kinase (A-kinase) is considered as a possible mechanism of insulin action for glycogen synthase activation. In this study, we investigated the time course of insulin action on human muscle A-kinase activity during a 2-h insulin infusion in 13 insulin-sensitive (group S) and 7 insulin-resistant subjects (group R). Muscle biopsies were obtained from quadriceps femoris muscle at times 0, 10, 20, 40, and 120 min. Insulin infusion resulted in significant inhibition of A-kinase activity at 20 and/or 40 min using 0.2, 0.6, and 1.0 microM cyclic AMP in group S. A-kinase activities both before and after insulin administration were lower in group S than in group R using 0.6 microM cyclic AMP. The decrease in apparent affinity for cyclic AMP during insulin infusion was larger for group S compared with group R. Glycogen synthase activity increased significantly after insulin infusion in both groups and was higher in group S compared with group R. The data suggest that a defective response of A-kinase to insulin in insulin-resistant subjects could contribute to their reduced insulin stimulation of skeletal muscle glycogen synthase.     

Studies on Urinary Phenolic Compounds in Man: II. Phenolic-Acids and -Amines During a Load of α-Methyl-Dopa and Disulfiram in Periodic Catatonia

A prolonged glucose load was administered to four patients with hypokalaemic periodic paralysis and four healthy control subjects. Muscle ATP and CP concentrations as well as lactate dehydrogenase, hexokinase and phosphorylase activities were similar in the two groups, but succinate dehydrogenase was approximately 50% higher in the control muscles. Muscle fibre composition was almost identical in the two groups, whereas patients had a higher degree of capillarization. Complete muscle weakness was produced in all patients, accompanied by hypokalaemia. Glucose loading resulted in elevated insulin levels and a minor rise in blood glucose level was seen in the patients compared to the control subjects. Glucose loading decreased hexokinase activity in controls, but increased this in the patients. At similar times, muscle and blood lactate levels and blood pyruvate values were generally higher in the patients over the course of the experiment. Initial glycogen concentrations were higher in patients, but glucose loading did not result in greatly increased glycogen values. These data suggest that patients with hypokalaemic periodic paralysis have an enhanced metabolism of carbohydrates and that insulin seems to be an important factor leading to the onset of muscle weakness.     

Effects of insulin infusion on human skeletal muscle pyruvate dehydrogenase, phosphofructokinase, and glycogen synthase. Evidence for their role in oxidative and nonoxidative glucose metabolism.

To determine whether activation by insulin of glycogen synthase (GS), phosphofructokinase (PFK), or pyruvate dehydrogenase (PDH) in skeletal muscle regulates intracellular glucose metabolism, subjects were studied basally and during euglycemic insulin infusions of 12, 30, and 240 mU/m2 X min. Glucose disposal, oxidative and nonoxidative glucose metabolism were determined. GS, PFK, and PDH were assayed in skeletal muscle under each condition. Glucose disposal rates were 2.37 +/- 0.11, 3.15 +/- 0.19, 6.71 +/- 0.44, and 11.7 +/- 1.73 mg/kg X min; glucose oxidation rates were 1.96 +/- 0.18, 2.81 +/- 0.28, 4.43 +/- 0.32, and 5.22 +/- 0.52. Nonoxidative glucose metabolism was 0.39 +/- 0.13, 0.34 +/- 0.26, 2.28 +/- 0.40, and 6.52 +/- 1.21 mg/kg X min. Both the proportion of active GS and the proportion of active PDH were increased by hyperinsulinemia. PFK activity was unaffected. Activation of GS was correlated with nonoxidative glucose metabolism, while activation of PDH was correlated with glucose oxidation. Sensitivity to insulin of GS was similar to that of nonoxidative glucose metabolism, while the sensitivity to insulin of PDH was similar to that of glucose oxidation. Therefore, the activation of these enzymes in muscle may regulate nonoxidative and oxidative glucose metabolism.