Impaired hepatic glycogen synthesis in glucokinase-deficient (MODY-2) subjects.

All glucokinase gene mutations identified to date have been localized to exons that are common to the pancreatic and hepatic isoforms of the enzyme. While impaired insulin secretion has been observed in glucokinase-deficient subjects the consequences of this mutation on hepatic glucose metabolism remain unknown. To examine this question hepatic glycogen concentration was measured in seven glucokinase-deficient subjects with normal glycosylated hemoglobin and 12 control subjects using 13C nuclear magnetic spectroscopy during a day in which three isocaloric mixed meals were ingested.

Glucose metabolism distal to a critical coronary stenosis in a canine model of low-flow myocardial ischemia.

Myocardial regions perfused through a coronary stenosis may cease contracting, but remain viable. Clinical observations suggest that increased glucose utilization may be an adaptive mechanism in such "hibernating" regions. In this study, we used a combination of 13C-NMR spectroscopy, GC-MS analysis, and tissue biochemical measurements to track glucose through intracellular metabolism in intact dogs infused with [1-13C]glucose during a 3-4-h period of acute ischemic hibernation.

Mechanism of free fatty acid-induced insulin resistance in humans.

To examine the mechanism by which lipids cause insulin resistance in humans, skeletal muscle glycogen and glucose-6-phosphate concentrations were measured every 15 min by simultaneous 13C and 31P nuclear magnetic resonance spectroscopy in nine healthy subjects in the presence of low (0.18 +/- 0.02 mM [mean +/- SEM]; control) or high (1.

The roles of insulin and glucagon in the regulation of hepatic glycogen synthesis and turnover in humans.

To determine the respective roles of insulin and glucagon for hepatic glycogen synthesis and turnover, hyperglycemic clamps were performed with somatostatin [0.1 micrograms/(kg.min)] in healthy young men under conditions of: (I) basal fasting) portal vein insulinemia-hypoglucagonemia, (II) basal portal vein insulinemia-basal glucagonemia, and (III) basal peripheral insulinemia-hypoglucagonemia.

Contribution of net hepatic glycogenolysis to glucose production during the early postprandial period.

Relative contributions of net hepatic glycogenolysis and gluconeogenesis to glucose production during the first 12 h of a fast were studied in 13 healthy volunteers by noninvasively measuring hepatic glycogen content using 13C nuclear magnetic resonance spectroscopy. Rates of net hepatic glycogenolysis were calculated by multiplying the change in liver glycogen content with liver volume determined by magnetic resonance imaging. Rates of gluconeogenesis were calculated as the difference between rates of glucose production determined with an infusion of [6,6-2H]-glucose and net hepatic glycogenolysis.

Direct assessment of liver glycogen storage by 13C nuclear magnetic resonance spectroscopy and regulation of glucose homeostasis after a mixed meal in normal subjects.

Despite extensive recent studies, understanding of the normal postprandial processes underlying immediate storage of substrate and maintenance of glucose homeostasis in humans after a mixed meal has been incomplete. The present study applied 13C nuclear magnetic resonance spectroscopy to measure sequential changes in hepatic glycogen concentration, a novel tracer approach to measure postprandial suppression of hepatic glucose output, and acetaminophen to trace the pathways of hepatic glycogen synthesis to elucidate the homeostatic adaptation to the fed state in healthy human subjects. After the liquid mixed meal, liver glycogen concentration rose from 207 +/- 22 to 316 +/- 19 mmol/liter at an average rate of 0.

Mass and positional isotopomer analysis of glucose metabolism in periportal and pericentral hepatocytes.

To determine whether the source of carbon for the indirect pathway of hepatic glycogen synthesis differs between the periportal and pericentral zones, we studied seven 24-h-fasted conscious rats given a constant 2-h intraduodenal infusion of glucose, 40% labeled with [U-13C]glucose (99% 13C enriched), to raise and maintain plasma glucose concentration at approximately 10 mM. Glycogen, glutamate, aspartate, and alanine were selectively sampled from the periportal and pericentral zones of the liver by the dual-digitonin pulse technique and analyzed by 13C-NMR for positional isotopomer distribution and by gas chromatography-mass spectrometry for mass isotopomer distribution. Plasma glucose mass isotopomer distribution was determined from gas chromatography-mass spectrometry.

Decreased muscle glucose transport/phosphorylation is an early defect in the pathogenesis of non-insulin-dependent diabetes mellitus.

Recent studies have demonstrated that reduced insulin-stimulated muscle glycogen synthesis is the major cause of insulin resistance in patients with non-insulin-dependent diabetes mellitus (NIDDM). This reduced rate has been assigned to a defect in either glucose transport or hexokinase activity. However it is unknown whether this is a primary or acquired defect in the pathogenesis of NIDDM.

Impaired net hepatic glycogen synthesis in insulin-dependent diabetic subjects during mixed meal ingestion. A 13C nuclear magnetic resonance spectroscopy study.

Hepatic glycogen concentration was measured in six subjects with insulin-dependent diabetes mellitus (IDDM) and nine weight-matched control subjects using 13C nuclear magnetic resonance spectroscopy during a day in which three isocaloric mixed meals were ingested. The relative fluxes of the direct and indirect (3 carbon units-->-->glycogen) pathways of hepatic glycogen synthesis were also assessed using [1-13C]glucose in combination with acetaminophen to noninvasively sample the hepatic UDP-glucose pool. Mean fasting hepatic glycogen content was similar in the two groups.

13C-nuclear magnetic resonance spectroscopy studies of hepatic glucose metabolism in normal subjects and subjects with insulin-dependent diabetes mellitus.

To determine the effect of insulin-dependent diabetes mellitus (IDDM) on rates and pathways of hepatic glycogen synthesis, as well as flux through hepatic pyruvate dehydrogenase, we used 13C-nuclear magnetic resonance spectroscopy to monitor the peak intensity of the C1 resonance of the glucosyl units of hepatic glycogen, in combination with acetaminophen to sample the hepatic UDP-glucose pool and phenylacetate to sample the hepatic glutamine pool, during a hyperglycemic-hyperinsulinemic clamp using [1-13C]-glucose. Five subjects with poorly controlled IDDM and six age-weight-matched control subjects were clamped at a mean plasma glucose concentration of approximately 9 mM and mean plasma insulin concentrations approximately 400 pM for 5 h. Rates of hepatic glycogen synthesis were similar in both groups (approximately 0.

Substrate cycling between pyruvate and oxaloacetate in awake normal and 3,3'-5-triiodo-L-thyronine-treated rats.

Substrate cycling between pyruvate and oxaloacetate was assessed in awake 24-h fasted normal and triiodothyronine (T3)-treated rats. After a 20- or 60-min infusion of [3-13C]alanine (99% enriched, 12 mg/min) the 13C enrichments of liver glucose and alanine carbons were analyzed by 13C and 1H nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry. Substrate cycling from phosphoenolpyruvate to pyruvate [via pyruvate kinase (PK)] and from oxaloacetate to pyruvate [via malic enzyme (ME)] relative to the pyruvate carboxylase (PC) flux [i.

Effects of puberty and diabetes on metabolism of insulin-sensitive fuels.

Insulin's ability to stimulate glucose metabolism is reduced during normal puberty; these changes are exaggerated in adolescents with insulin-dependent diabetes mellitus (IDDM). Because the effects of puberty and IDDM on the other actions of insulin have not been established, we studied leucine kinetics (using [1-13C]leucine) and fat metabolism during euglycemic hyperinsulinemia (20 mU.m2.

Liver glycogen turnover in fed and fasted humans.

Whether liver glycogen synthesis and breakdown occur simultaneously during net glycogen synthesis was assessed in fed and fasted healthy humans. The peak intensity of the carbon-1 (C1) resonance of the glycosyl units of glycogen was monitored with 13C nuclear magnetic resonance spectroscopy during [1-13C]glucose infusion followed by unlabeled glucose infusion. The C1 peak intensity increased almost linearly during the [1-13C]glucose infusion, reflecting a near linear rate of glycogen synthesis.

Effect of hepatic nerves on disposition of an intraduodenal glucose load.

We examined the disposition of a continuous 4-h intraduodenal glucose infusion (8 mg.kg-1 x min-1, labeled with [1-13C]glucose and [3-3H]glucose) in nine conscious hepatic-denervated dogs. Cumulative net hepatic uptakes (in grams of glucose equivalents) were 13.

1% Cyclopentolate hydrochloride: another look at the time course of cycloplegia using an objective measure of the accommodative response.

The time course of cycloplegia was measured by monitoring residual accommodation after the application of 1 drop (29.3 microliters) of 1% cyclopentolate hydrochloride. Three different measures of residual accommodation were made, one objective assessment with an optometer, and two subjective assessments similar to those used by previous investigators.

Comparative effects of monomethylsuccinate and glucose on insulin secretion from perifused rat islets.

In the absence of any other exogenously added fuel, monomethylsuccinate, the methyl ester of succinic acid, at 10-20 mM stimulates insulin release in a biphasic pattern. In quantitative terms, first-phase release evoked by 20 mM MMSucc was comparable to that observed with 20 mM glucose but second-phase release was only 20% of the glucose-induced response. Secretion to both MMSucc and glucose was virtually abolished by the calcium channel antagonist nitrendipine (0.

Enhancement of the gluconeogenic flux of hepatic glycogen repletion by a phenacyl imidazolium compound in vivo.

The effect of a phenacyl imidazolium compound (LY177507 or Proglycosyn, Eli Lilly) on the direct (glucose-->glucose-6-phosphate-->glycogen) and indirect (three-carbon compounds-->glucose-6-phosphate-->glycogen) pathways of liver glycogen synthesis was studied in conscious rats. [1-13C]Glucose (99% enriched) was infused intraduodenally into chronically catheterized Proglycosyn-treated (n = 7) and saline-treated (n = 7) rats for 120 min. Net hepatic glycogen synthetic rates were increased twofold in drug-treated rats compared with saline-treated controls.

Sources of carbon for hepatic glycogen synthesis in the conscious dog.

To identify the source(s) of carbon for the indirect pathway of hepatic glycogen synthesis, we studied nine 42-h fasted conscious dogs given a continuous intraduodenal infusion of glucose, labeled with [1-13C]glucose and [3-3H]glucose, at 8 mg.kg-1.min-1 for 240 min.

Quantitative analysis of the pathways of glycogen repletion in periportal and perivenous hepatocytes in vivo.

In order to examine the pathways of hepatic glycogen repletion in the periportal and perivenous zones of the liver, [1-13C]glucose (99% enriched) was infused intraduodenally into conscious, 24-h fasted rats for 3 h. The liver was then quickly perfused in situ, and the cytoplasmic contents of the periportal and perivenous hepatocytes were selectively sampled by modification of the dual-digitonin-pulse technique (Quistorff, B., and Grunnet, N.

Quantitative comparison of pathways of hepatic glycogen repletion in fed and fasted humans.

The effect of fasting vs. refeeding on hepatic glycogen repletion by the direct pathway, i.e.

Effects of a hydroxamate siderophore on iron absorption by sunflower and sorghum.

When Fe was supplied at 100 micromolar in nutrient solution of pH 7.5, 10 and 1 micromolar levels of the siderophore desferrioxamine B (DFOB), a microbial iron transport compound, significantly (alpha = 0.05) enhanced growth and reduced chlorosis of an Fe-inefficient variety of sorghum (Sorghum bicolor L.

The properties of immune complex-forming systems. A new theoretical approach.

A new mathematical model for antigen-antibody interactions has been developed. The new model is based on the assumption that the formation of complexes between a bivalent antibody and a multivalent antigen is determined thermodynamically by the concentrations and valences of antigen as well as antibody, together with one association constant which is common to all mutual interactions. Formulae have been derived for calculation of the distributions of compositionally different antigen-antibody complexes either from knowledge of equilibrium concentrations of free antigen and antibody, or from knowledge of total amounts of antigen and antibody in the system.