The metabolic syndrome and the heart--a considered opinion.

The metabolic syndrome (MS) is a multifactorial, heterogeneous group of risk factors for the development of cardiovascular disease. Here we review the evidence in support of the hypothesis that metabolic dysregulation of the body as a whole leads to contractile dysfunction of the heart due to an imbalance of substrate uptake (increased) and substrate oxidation (decreased). The consequences of this imbalance were already recognized 150 years ago by Virchow when he described "fatty atrophy" of the heart as a "true metamorphosis of the heart muscle cell."

Ingested IFN-alpha preserves residual beta cell function in type 1 diabetes.

Type 1 diabetes mellitus is a chronic disorder that presumably results from an autoimmune destruction of the insulin-producing pancreatic beta cells. The therapeutic potential of interventions aimed at preventing type 1 diabetes can be assessed in newly diagnosed patients. Because there is a historical experience of a low incidence of spontaneous remission in type 1 diabetes mellitus, interventions preserving beta cell function have been used to identify positive therapeutic outcomes. We treated 10 newly diagnosed type 1 diabetes patients with 30,000 IU ingested interferon-alpha (IFN-alpha) within 1 month of diagnosis and examined the difference between baseline and Sustacal-induced (Mead Johnson Nutritionals, Evansville, IN) C-peptide responses, respectively, at 0, 3, 6, 9, and 12 months. Eight of the ten patients showed preserved beta cell function, with at least a 30% increase in stimulated C-peptide levels at 0, 3, 6, 9, and 12 months after initiation of treatment. There was no discernible chemical or clinical toxicity associated with ingested IFN-alpha. Our results support the potential of ingested IFN-alpha to preserve residual beta cell function in recent onset type 1 diabetes mellitus and the testing of IFN-alpha in a placebo-controlled trial.

Evidence that forskolin binds to the glucose transporter of human erythrocytes.

Binding of [4-3H]cytochalasin B and [12-3H]forskolin to human erythrocyte membranes was measured by a centrifugation method. Glucose-displaceable binding of cytochalasin B was saturable, with KD = 0.11 microM, and maximum binding approximately 550 pmol/mg of protein. Forskolin inhibited the glucose-displaceable binding of cytochalasin B in an apparently competitive manner, with K1 = 3 microM. Glucose-displaceable binding of [12-3H]forskolin was also saturable, with KD = 2.6 microM and maximum binding approximately equal to 400 pmol/mg of protein. The following compounds inhibited binding of [12-3H]forskolin and [4-3H]cytochalasin B equivalently, with relative potencies parallel to their reported affinities for the glucose transport system: cytochalasins A and D, dihydrocytochalasin B, L-rhamnose, L-glucose, D-galactose, D-mannose, D-glucose, 2-deoxy-D-glucose, 3-O-methyl-D-glucose, phloretin, and phlorizin. A water-soluble derivative of forskolin, 7-hemisuccinyl-7-desacetylforskolin, displaced equivalent amounts of [4-3H]cytochalasin B or [12-3H]forskolin. Rabbit erythrocyte membranes, which are deficient in glucose transporter, did not bind either [4-3H]cytochalasin B or [12-3H]forskolin in a glucose-displaceable manner. These results indicate that forskolin, in concentrations routinely employed for stimulation of adenylate cyclase, binds to the glucose transporter. Endogenous ligands with similar specificities could be important modulators of cellular metabolism.

Cyclic AMP does not modulate hexose uptake of serum-deprived BHK-21 fibroblasts.

We have studied the uptakes of the glucose analogs, 2-deoxyglucose (2DG), and 3-O-methylglucose (3OMG) in serum-deprived BHK-21 fibroblasts. Incubation for 4 hours with 0.25 mM 1-methyl-3-isobutylxanthine (MIX) increased cellular cyclic AMP 3 to 15 fold, without affecting the initial rates of uptake of these sugars. Incubation with 100 nM insulin, doubled hexose uptake without affecting cyclic AMP. Insulin did not modify the cyclic AMP response to MIX; nor did MIX alter the hexose uptake response to insulin. There was no effect of 0.1 mM L-isoproterenol (ISO) or 0.1 mM prostaglandin E1 (PGE1) on cyclic AMP. PGE1 slightly stimulated uptake of 2DG but not 3OMG; ISO did not affect hexose uptake. We conclude that cyclic AMP does not directly regulate hexose uptake in these cells.

Effects of insulin on glycogen metabolism of serum-deprived BHK-21 fibroblasts.

We have studied the effects of insulin on glycogen metabolism of cultured BHK-21 fibroblasts. Addition of insulin to cells cultured in 0.5% serum stimulated incorporation of 5mM 14C-glucose into glycogen and increased cellular glycogen content, without inducing proliferation. With serum-deprived cells incubated for 2 hr, maximal stimulation of incorporation of glucose into glycogen was 3.5-fold and the half-maximal dose of insulin was 5nM. After culture for 24 hr with 100nM insulin, incorporation was increased 13-fold and glycogen content was increased by 44%. Incorporation of glucose into glycogen was reduced by agents which elevate cellular adenosine 3',5-cyclic monophosphate (cyclic AMP): 10 microM prostaglandin E1 (PGE1), 0.25 mM 1-methyl-3-isobutylxanthine (MIX), 10 microM L-epinephrine (L-EPI), and 10 microM L-isoproterenol (L-ISO). Culture with 100nM insulin for 2 hr stimulated incorporation three-fold in the presence of any of these compounds, but insulin did not affect cellular cyclic AMP. At all times from 1 hr to 24 hr after addition of insulin to serum-deprived cultures, the 1-min uptake of 10 microM 3H-2-deoxyglucose (3H-2DG) was increased. There were small (30 to 40%) increases in glycogen synthase I activity at 15 and 30 min but not 4 hr after addition of insulin. After 24 hr with insulin, total glycogen synthase and phosphorylase activities were increased approximately two-fold, without changes in their activation states. We conclude that insulin promotes glycogenesis in serum-deprived BHK-21 cells. This response is mediated principally by increased entry of glucose into cells, and is not mediated by a change in cellular cyclic AMP concentration.

[12-Homoarginine]glucagon: synthesis and observations on conformation, biological activity, and copper-mediated peptide cleavage.

Specific modification of the single lysine residue (Lys-12) in glucagon with O-methylisourea has been effected by blocking the reactivity of the amino terminal histidine with copper, providing a method for obtaining [12-homoarginine]glucagon. It was found that as a side reaction, under the conditions of the modification reaction, Cu(II) catalyzed cleavage of the polypeptide chain between Asp-9 and Tyr-10, and between Lys-12 and Tyr-13. This observation may be of value for development of a sequence-specific peptide cleavage procedure. The dilute solution conformations of glucagon and [12-homoarginine]-glucagon were compared by circular dichroism, fluorescence, phosphorescence, energy transfer, and optical detection of magnetic resonance. The results indicate that conversion of Lys-12 to homoarginine does not alter the helix content the side chain conformation in the vicinity of the tyrosine and tryptophan residues, or the relative distances and orientations between these residues. However, the modification reduces the hormone potency towards activation of lipolysis in isolated rat epididymal fat cells by a factor of seven. We attribute the loss of potency to an interference with a specific interaction between the lysine residue and the fat cell hormone receptor, and not to a change in the solution conformation of the hormone.

Hexokinase isozymes of normal human subcutaneous adipose tissue.

The isozymes of hexokinase in surgical specimens of human subcutaneous adipose tissue were separated by elution from DEAE-cellulose with linear KCl gradients at ph 7.4. Two peaks of activity were found: Peak 1 eluted at 0.05M KCl, and Peak 2 at 0.19M KCl. Michaelis constants (Km) for glucose were: Peak 1, 6.5 x 10-5M; Peak 2, 1.5 x 10-4M. Peak 2 was more susceptible than Peak 1 to inactivation by trypsin, 0.1 mg/ml, and was protected by 0.1M glucose. Both peaks were protected from heat inactivation (45 degrees) by 0.1M glucose. Peak 2 comprised 66 +/- 5 percent of the total hexokinase activity. No activity with the characteristics of hexokinase III was detected in human fat. In all these characteristics, the isozymes of human adipose tissue closely resemble hexokinases I and II from rats.

Comparison of the lipolytic effects of insulin and proinsulin on isolated fat cells.

Proinsulin, like insulin, has a biphasic effect on the lipolysis of isolated fat cells. Low concentrations of the peptides inhibit the response to submaximal doses of epinephrine, while higher concentrations of the peptides enhance the effect of supramaximal doses of epinephrine. Half-maximal doses are: (a) inhibition of lipolysis: insulin, 2.3 times 10- minus 11M; proinsulin, 6.9 times 10- minus 10M; (b) enhancement of lipolysis: insulin, 3.1 times 10- minus 9M; proinsulin, 8.1 times 10- minus 8M. The ratios of the potencies of the peptides are the same for both effects. These results suggest that both the inhibitory and the lipolytic effects of insulin are not caused by contaminants, and are mediated by similar receptors.