Prandial glucose effectiveness and fasting gluconeogenesis in insulin-resistant first-degree relatives of patients with type 2 diabetes.
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Impaired glucose effectiveness (i.e., a diminished ability of glucose per se to facilitate its own metabolism), increased gluconeogenesis, and endogenous glucose release are, together with insulin resistance and beta-cell abnormalities, established features of type 2 diabetes. To explore aspects of the pathophysiology behind type 2 diabetes, we assessed in a group of healthy people prone to develop type 2 diabetes (n = 23), namely first-degree relatives of type 2 diabetic patients (FDR), 1) endogenous glucose release and fasting gluconeogenesis measured using the 2H2O technique and 2) glucose effectiveness. The FDR group was insulin resistant when compared with an age-, sex-, and BMI-matched control group without a family history of type 2 diabetes (n = 14) (M value, clamp: 6.07 +/- 0.48 vs. 8.06 +/- 0.69 mg x kg(-1) lean body weight (lbw) x min(-1); P = 0.02). Fasting rates of gluconeogenesis (1.28 +/- 0.06 vs. 1.41 +/- 0.07 mg x kg(-1) lbw x min(-1); FDR vs. control subjects, P = 0.18) did not differ in the two groups and accounted for 53 +/- 2 and 60 +/- 3% of total endogenous glucose release. Glucose effectiveness was examined using a combined somatostatin and insulin infusion (0.17 vs. 0.14 mU x kg(-1) x min(-1), FDR vs. control subjects), the latter replacing serum insulin at near baseline levels. In addition, a 360-min labeled glucose infusion was given to simulate a prandial glucose profile. After glucose infusion, the integrated plasma glucose response above baseline (1,817 +/- 94 vs. 1,789 +/- 141 mmol/l per 6 h), the ability of glucose to simulate its own uptake (1.50 +/- 0.13 vs. 1.32 +/- 0.16 ml x kg(-1) lbw x min(-1)), and the ability of glucose per se to suppress endogenous glucose release did not differ between the FDR and control group. In conclusion, in contrast to overt type 2 diabetic patients, healthy people at high risk of developing type 2 diabetes are characterized by normal glucose effectiveness at near-basal insulinemia and normal fasting rates of gluconeogenesis. (+info)
Gluconeogenesis in moderately and severely hyperglycemic patients with type 2 diabetes mellitus.
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We tested the generally accepted concept that increased gluconeogenesis (GNG) and endogenous glucose production (EGP) are the main reasons for postabsorptive hyperglycemia in patients with type 2 diabetes mellitus (T2DM). GNG was measured with the (2)H(2)O method by use of both the C5-to-C2 ratio (C5/C2, with gas chromatography-mass spectrometry) and the C5-to-(2)H(2)O ratio (C5/(2)H(2)O, with isotope ratio mass spectrometry), and EGP was measured with 3-[(3)H]glucose in 27 patients with T2DM [13 with fasting plasma glucose (FPG) >10 mM and 14 with FPG <10 mM] and in 7 weight- and age-matched nondiabetic controls. The results showed 1) that GNG could be determined accurately with (2)H(2)O by using either C5/C2 or C5/(2)H(2)O; 2) that whereas after an overnight fast of 16 h, GNG was higher in the entire group of patients with T2DM than in controls (6.4 vs. 5.0 micromol. kg(-1). min(-1) or 60.4 vs. 51.4% of EGP, P < 0.02), GNG was within normal limits (less than the mean +/- 2 SD of controls or <65.3%) in 11/14 (79%) patients with mild to moderate hyperglycemia (FPG <10 mM) and in 5/13 (38%) of patients with severe hyperglycemia (FPG 10-20 mM); 3) that elevated GNG in T2DM was associated with a 43% decrease in prehepatic insulin secretion, i.e., with hepatic insulin deficiency; and 4) that FPG correlated significantly with glucose clearance (insulin resistance) (r = 0.70) and with GNG (r = 0.50) or EGP (r = 0.45). We conclude 1) that peripheral insulin resistance is at least as important as GNG (and EGP) as a cause of postabsorptive hyperglycemia in T2DM and 2) that GNG and EGP in T2DM are increased under conditions of significant hepatic insulin deficiency and thus probably represent a late event in the course of T2DM. (+info)
Phosphoenolpyruvate synthetase from the hyperthermophilic archaeon Pyrococcus furiosus.
(75/1508)
Phosphoenolpyruvate synthetase (PpsA) was purified from the hyperthermophilic archaeon Pyrococcus furiosus. This enzyme catalyzes the conversion of pyruvate and ATP to phosphoenolpyruvate (PEP), AMP, and phosphate and is thought to function in gluconeogenesis. PpsA has a subunit molecular mass of 92 kDa and contains one calcium and one phosphorus atom per subunit. The active form has a molecular mass of 690+/-20 kDa and is assumed to be octomeric, while approximately 30% of the protein is purified as a large ( approximately 1.6 MDa) complex that is not active. The apparent K(m) values and catalytic efficiencies for the substrates pyruvate and ATP (at 80 degrees C, pH 8.4) were 0.11 mM and 1.43 x 10(4) mM(-1). s(-1) and 0.39 mM and 3.40 x 10(3) mM(-1) x s(-1), respectively. Maximal activity was measured at pH 9.0 (at 80 degrees C) and at 90 degrees C (at pH 8.4). The enzyme also catalyzed the reverse reaction, but the catalytic efficiency with PEP was very low [k(cat)/K(m) = 32 (mM. s(-1)]. In contrast to several other nucleotide-dependent enzymes from P. furiosus, PpsA has an absolute specificity for ATP as the phosphate-donating substrate. This is the first PpsA from a nonmethanogenic archaeon to be biochemically characterized. Its kinetic properties are consistent with a role in gluconeogenesis, although its relatively high cellular concentration ( approximately 5% of the cytoplasmic protein) suggests an additional function possibly related to energy spilling. It is not known whether interconversion between the smaller, active and larger, inactive forms of the enzyme has any functional role. (+info)
Hepatic phosphoenolpyruvate carboxykinase gene expression is not repressed by dietary carbohydrates in rainbow trout (Oncorhynchus mykiss).
(76/1508)
Phosphoenolpyruvate carboxykinase (PEPCK) is a rate-limiting enzyme in hepatic gluconeogenesis and therefore plays a central role in glucose homeostasis. The aim of this study was to analyse the nutritional regulation of PEPCK gene expression in rainbow trout (Oncorhynchus mykiss), which are known to use dietary carbohydrates poorly. A full-length hepatic PEPCK cDNA (2637 base pairs with one open reading frame putatively encoding a 635-residue protein) was cloned and found to be highly homologous to mammalian PEPCKs. The presence of a putative peptide signal specific to a mitochondrial-type PEPCK in the deduced amino acid sequence suggests that this PEPCK gene codes for a mitochondrial form. In gluconeogenic tissues such as liver, kidney and intestine, this PEPCK gene was expressed at high levels and, in the liver we found no regulation of PEPCK gene expression by dietary carbohydrates. These results suggest that the first step of the hepatic gluconeogenic pathway in rainbow trout is functional and highly active irrespective of the dietary carbohydrate supply. (+info)
Influence of increasing carbohydrate intake on glucose kinetics in injured patients.
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The metabolic and hormonal effect of glucose loads, ranging from 125 to 504 g/70 kg/day, were studied in severely injured patients. There was little or no correlation of glucose intake with nitrogen balance, plasma glucose, fatty acid concentrations, or epinephrine excretion. Increased norepinephrine excretion correlated with and may have resulted from increased glucose intake. Serum glucagon concentrations averaged 320 pg/ml and were not depressed by glucose intake. Insulin concentrations rose with glucose intake but were low for the level of plasma glucose. Glucose oxidation and non-oxidative metabolism, including glycogen deposition, correlated well with glucose intake. Gluconeogenesis from alanine was much higher than normal but was completely suppressed at very high intakes. The data imply that cycling of glucose, with glycerol, glycogen, or both, increased with increasing glucose intake. (+info)
Gluconeogenesis from L-cysteine in the perfused rat liver.
(78/1508)
The effects of dietary and hormonal treatments on the rate of gluconeogenesis from L-cysteine have been investigated in the perfused rat liver in situ. In order to demonstrate gluconeogenesis from L-cysteine, rats were fed either a 90% casein diet, or this diet with 2 or 4% cysteine, added in place of casein, and perfused in the fed state; or fed stock diet and starved 48 or 72 hours; or fasted and injected with cortisol. The net rate of gluconeogenesis (in mumoles/min/g liver) from cysteine in rats fed 4%cysteine was 0.24; in 48-hour starved rats it was 0.10; in 72-hour starved rats it was 0.16; and in the cortisol injected rats it was 0.23. When [U-14C]cysteine plus carrier cysteine (10 mM) was added as the substrate for gluconeogenesis in 72-hour starved rats; 3.9% of the label appeared in glucose. The above dietary and hormonal treatments stimulated gluconeogenesis from L-cysteine. (+info)
Hypoglycemia induced by insulin increases hepatic capacity to produce glucose from gluconeogenic amino acids.
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AIM: To investigate the hepatic capacity to produce glucose during hypoglycemia induced by insulin (HII). METHODS: Livers from 24-h fasted rats which received i.p. insulin (HII rats) or saline (control rats) were perfused in situ. The gluconeogenic substrates L-alanine (5 mmol/L), L-glutamine (5 mmol/L), L-lactate (2 mmol/L), and glycerol (2 mmol/L) were employed. The gluconeogenic activity was measured as the difference between rates of glucose released during and before the substrate infusion. In part of the experiments the production of urea was measured. Before the liver perfusion blood was collected for determination of glycemia and insulinemia. RESULTS: HII rats showed: (a) hypoglycemia and hyperinsulinemia; (b) increased hepatic capacity to produce glucose from L-alanine and L-glutamine; (c) increased hepatic ureogenesis from L-alanine and L-glutamine; and (d) increased hepatic glucose production from glycerol. However, hepatic glucose production from L-lactate was not affected by hypoglycemia. CONCLUSION: In spite of hyperinsulinemia the hepatic capacity to produce glucose from L-glutamine and L-alanine increased during HII. These results can be attributed to the higher hepatic catabolism of both amino acids, since the ability of the liver to produce glucose was not affected by hypoglycemia. (+info)
Modulation by cyclic AMP and phorbol myristate acetate of cephaloridine-induced injury in rat renal cortical slices.
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Intracellular signaling pathways of cAMP and protein kinase C (PKC) have been suggested to modulate the generation of free radicals. We investigated the effects of cAMP and phorbol myristate acetate (PMA), a PKC activator, on cephaloridine (CER)-induced renal cell injury, which has been reported to be due to the generation of free radicals. Incubation of rat renal cortical slices with CER resulted in increases in lipid peroxidation and lactate dehydrogenase (LDH) release and in decreases in gluconeogenesis and p-aminohippurate (PAH) accumulation in rat renal cortical slices, suggesting free radical-induced injury in slices exposed to CER. A derivative of cAMP ameliorated not only the increase in lipid peroxidation but also the renal cell damage induced by CER. This amelioration by a cAMP derivative of lipid peroxidation and renal cell damage caused by CER was blocked by KT 5720, a protein kinase A (PKA) inhibitor. Lipid peroxidation and the indices of cell injury were increased by PMA. PMA also enhanced CER-induced lipid peroxidation and cell damage in the slices. This enhancement by PMA of CER-induced injury was blocked by H-7, a PKC inhibitor. These results indicated that intracellular signaling pathways of cAMP and PKC modulate free radical-mediated nephrotoxicity induced by CER. (+info)