Temporal optimization of exercise to lower fasting glucose levels.

Exercise stimulates glucose uptake and increases insulin sensitivity acutely. Temporally optimizing exercise timing may minimize the nocturnal rise in glucose levels. This study examined the effect of exercise timing on evening and overnight glucose concentrations in individuals who were non-obese with normal fasting glucose levels (Non-Ob; n = 18) and individuals with obesity (OB) with impaired fasting glucose levels (OB+IFG) and without (n = 16 and n = 18, respectively). Subjects were studied on three occasions (no exercise (NOEX)), morning exercise (AMEX; 0700 h) and evening exercise (PMEX; 2000 h). The evening meal was provided (1800 h) and blood samples were taken from 1740 to 0700 h and morning endogenous glucose production (EGP) was measured. Glucose and insulin concentrations increased with the dinner meal with peak concentrations being higher in OB+IFG than in OB and Non-Ob (P = 0.04). In OB+IFG, evening glucose concentrations rose above baseline levels at about 2300 h, with the glucose concentrations staying somewhat lower with AMEX and PMEX until ∼0500 h than with NOEX. In OB+IFG, insulin concentrations decreased following the dinner meal and waned throughout the night, despite the rising glucose concentrations. In the OB and Non-Ob individuals following the dinner meal, no increase in glucose concentrations occurred in the evening period and insulin levels mirrored this. No difference was observed in the morning fasting glucose levels between study days or between groups. Regardless of time of day, exercise delays the evening rise in glucose concentrations in adults with OB+IFG but does not lower morning fasting glucose levels or improve the synchrony between glucose and insulin concentrations. KEY POINTS: Insulin resistance and type 2 diabetes have been linked to disturbances of the core clock, and glucose tolerance demonstrates a diurnal rhythm in healthy humans with better glucose tolerance in the morning than in the afternoon and evening. Skeletal muscle is a primary site for insulin resistance in people with impaired glucose tolerance. In individuals with obesity and impaired fasting glucose levels (OB+IFG), following a dinner meal, glucose concentrations started to rise and continues throughout the night, resulting in elevated glucose levels, while concomitantly, insulin levels are waning. Exercise, regardless of the time of day, suppressed the rise in glucose levels in OB+IFG for many hours during the night but did not lower morning fasting glucose levels. Morning exercise was not quite as effective as evening exercise.

Emerging marine diseases--climate links and anthropogenic factors.

Mass mortalities due to disease outbreaks have recently affected major taxa in the oceans. For closely monitored groups like corals and marine mammals, reports of the frequency of epidemics and the number of new diseases have increased recently. A dramatic global increase in the severity of coral bleaching in 1997-98 is coincident with high El Niño temperatures. Such climate-mediated, physiological stresses may compromise host resistance and increase frequency of opportunistic diseases. Where documented, new diseases typically have emerged through host or range shifts of known pathogens. Both climate and human activities may have also accelerated global transport of species, bringing together pathogens and previously unexposed host populations.

Hurricane Allen's Impact on Jamaican Coral Reefs.

Coral reefs of north Jamaica, normally sheltered, were severely damaged by Hurricane Allen, the strongest Caribbean hurricane of this century. Immediate studies were made at Discovery Bay, where reef populations were already known in some detail. Data are presented to show how damage varied with the position and orientation of the substraturn and with the shape, size, and mechanical properties of exposed organisms. Data collected over succeeding weeks showed striking differences in the ability of organisms to heal and survive.

Degradation of pigeon liver fatty acid synthetase in the absence of exogenous proteinases.

The homogeneity of pigeon liver fatty acid synthetase has been rigorously tested by physicochemical techniques and crossed-rocket immunoelectrophoresis. The enzyme has also been incubated for 1 h at 100 degrees C in 2% sodium dodecyl sulfate and 0.1 M dithiothreitol. The number of protein components on gel electrophoresis and of dansylated amino acids increased as a function of incubation time. Furthermore, the minor proteins observed after gel electrophoresis cross-reacted with antibody raised to the synthetase. Proteolysis was not chemically mediated by the detergent, the reducing agent or the buffer conditions chosen. Several commercially prepared proteins were not degraded by this procedure, and two proteins were recalcitrant to hydrolysis when included in the same incubation mixture as the synthetase. The inclusion of certain microbial proteinase inhibitors decreased the amount of degradation. This demonstrated that hydrolysis of the synthetase is mediated by a specific vertebrate enzyme which retains activity under denaturing conditions at 100 degrees C. Further degradation is also observed after individual treatment of four limited digestion products from the pigeon liver fatty acid synthetase, suggesting the possibility of an inherent proteolytic activity within the complex.

Isolation and characterization of an acyl carrier protein from pigeon liver fatty acid synthetase by controlled proteolysis with elastase.

Controlled proteolytic cleavage of 4'-phospho[14C]pantetheine-labeled pigeon liver fatty acid synthetase generates two 4'-phospho[14C]pantetheine-labeled peptides, Ec1 and Ec2. These are separated from each other and the core enzyme by gel permeation chromatography on a Sephadex G-75 column. The two radioactively labeled peptides constitute 50% of the radioactivity initially present in the 4'-phospho[14C]pantetheine-labeled fatty acid synthetase. The remaining label in the core enzyme is released quantitatively by proteolytic cleavage with trypsin. The molecular weights of Ec1 and Ec2 peptides, as determined by size exclusion chromatography and SDS-polyacrylamide gel electrophoresis, are 12000 and 6000, respectively. Both the higher and lower molecular weight peptides are homogeneous with respect to size and charge, as shown by polyacrylamide gel electrophoresis in the presence and absence of SDS. The higher molecular weight peptide, Ec1, is characterized as an acyl carrier protein by the transacylation reaction between the unlabeled Ec1 peptide and radioactively labeled acetyl- and malonyl-CoA. Since Ec2 peptide also contains the prosthetic group present in the Ec1 peptide, the Ec2 peptide appears to result from the proteolytic cleavage of the higher molecular weight peptide, Ec1. Amino acid composition of the acyl carrier protein shows the presence of 1 mol of 4'-phosphopantetheine per mol of protein. 2 mol of acyl carrier protein are present per mol of the fatty acid synthetase. The amino acid analysis is in good agreement with the molecular weight of the Ec1 peptide, as determined by gel filtration and SDS-polyacrylamide gel electrophoresis. N-Terminal amino acid analysis of this peptide shows the presence of an arginine residue.

An improved purification and further characterization of the messenger ribonucleic acid for the fatty acid synthetase from rat liver.

High purity fatty acid synthetase mRNA has been prepared from rat liver. The translational purity of the mRNA preparation was at least 27% as judged by the percentage of the radioactivity incorporated into acid-insoluble material that was precipitated by anti-fatty acid synthetase antibody. The specific activity of the mRNA was 220-times greater than that reported previously from this laboratory [1]. The large increase in the specific activity was achieved by the repeated use of high resolution linear-log sucrose density gradient centrifugation and the removal of 28 S rRNA by Sepharose 4B chromatography, as well as by the optimization of the K+ concentration (160 mM) in the reticulocyte lysate translation system. The mRNA preparation showed a single major band on agarose gel electrophoresis under denaturing conditions, and the translational activity of the fatty acid synthetase mRNA on the gel was found to coincide with this band. The molecular weight of the fatty acid synthetase mRNA is 2.5.10(6) Da. The mRNA directed the synthesis of fatty acid synthetase with a molecular weight indistinguishable from that of the authentic enzyme subunit (Mr = 240 000). The copurification of the translation product and authentic enzyme revealed that the fatty acid synthetase polypeptides synthesized in the reticulocyte lysate system are assembled in vitro into dimers, the native form of the enzyme.

Liberation, purification, and properties of thioesterase component of pigeon liver fatty acid synthetase complex.

Proteolysis of pigeon liver fatty acid synthetase with elastase results in the quantitative cleavage of the thioesterase component from the enzyme complex. This thioesterase component is two or three times more active catalytically in the isolated state than in the native fatty acid synthetase, and its activity is not affected by the presence or absence of reducing thiols. The proteolytically cleaved thioesterase is separated from the core enzyme in one step by size-exclusion chromatography on a Sephadex G-75 column. The peptide obtained by gel permeation is homogeneous with respect to size and charge, as shown by polyacrylamide gel electrophoresis in the presence and absence of SDS. Size-exclusion chromatography on Bio-Gel A 0.5 m and Sephadex G-75 columns, sucrose density gradient ultracentrifugation, and N-terminal amino acid analysis also indicate that the proteolytically cleaved thioesterase is homogeneous. The sedimentation coefficient of the thioesterase is approximately 2.9 S. Proteolytic cleavage with elastase also quantitatively releases the [1,3-14C]- or [1,3-3H]diisopropylphosphofluoridate-labeled thioesterase component from the correspondingly labeled fatty acid synthetase. Binding studies with 14C- or 3H-labelled diisopropylphosphofluoridate and fatty acid synthetase show that 2 mol of the label are bound per mol of the enzyme when complete loss of fatty acid-synthesizing activity occurs. The molecular weight of the thioesterase component is estimated to be 36000 by size-exclusion chromatography, SDS-polyacrylamide gel electrophoresis and amino acid analysis.

Comparative atherogenic effects of cholesterol and cholesterol oxides.

Previous findings indicating that the oxidation products of cholesterol are associated with atherogenicity have led to a comparative study of the subchronic effects of feeding rabbits purified cholesterol, oxidized cholesterols free of cholesterol and cholesterol esters, or a mixture of cholesterol and oxidized cholesterols. Macroscopically, the cholesterol-fed animals exhibited 6-fold more arterial lesions than the animals fed cholesterol-free oxidized cholesterols. Microscopically, there was no statistically significant difference from the control in the number of histochemically-defined lesions in any of the groups. However, the lesions in the cholesterol-fed group were more severe, as indicated by a statistically significant increase in the magnitude of the lesions. This increased severity was also characterized by greater frequency and intensity of Azure A/Thionin, VonKossa, and Horseradish Peroxidase-Wheat Germ Agglutinin staining. Electron-microscopic studies of normal appearing arterial tissues showed an increased density of viable smooth muscle cells and an increase in vacuolar extracellular debris in the cholesterol-fed group. Oxidized cholesterols in the concentrations and relative compositions administered here are markedly less atherogenic to rabbits than highly purified cholesterol.

The influence of dietary unsaturated cis and trans and saturated fatty acids on tissue lipids of swine.

A feeding trial was conducted to evaluate the effects of dietary trans unsaturated fatty acids (trans fat) and of the interplay of dietary saturated fatty acids (saturated fat), cis unsaturated fatty acids, (cis fat) and trans fat on tissue lipids, particularly those effects suggestive of angiotoxicity. Swine were fed for 10 months a diet containing 17% added fat. Seven blends of varying proportions of the 3 fat components provided sufficient sample points to permit an examination of the interplay. Parameters under study included weight gain, serum cholesterol and triglyceride concentrations, lipoprotein lipid profile, total lipid and cholesterol concentrations of liver, heart and aorta, fatty acid composition of liver and aorta lipids and hepatic fatty acid synthesis and cholesterol synthesis and oxidation. Fat blends containing disproportionately high levels of saturated or cis fat generally elicited responses consistent with results reported by others. The notable exception was the serum cholesterol concentration. Throughout the study, the swine were hypercholesterolemic. Swine fed the high saturated fat blend had serum cholesterol levels equal to those swine fed the high cis fat blend. Serum cholesterol levels in the swine fed the other fat blends were more elevated. Another apparent anomaly was the lower concentration of lipid in the aortas of swine fed the high-saturated fat diet. The impact of the trans fat was modulated by the relative proportions of saturated and cis fat in the diet. The impact of trans fat was of greater magnitude for most parameters when the fat blend was low in saturated fat. The sole parameter suggestive of trans fat-mediated angiotoxicity was the distribution of lipids in lipoprotein fractions. Swine fed diets containing trans fat had lower relative proportions of the alpha-lipoprotein lipids. Although hypercholesterolemic, the high fat diets were not overtly angiotoxic except when fed to swine that carried a specific immunogenetically-defined low density lipoprotein.

De novo fatty acid synthesis and fatty acid elongation catalyzed by subcellular fractions from hog and human aorta.

De novo synthesis and mitochondrial elongation of fatty acids have been demonstrated in subcellular fractions from hog and human aorta. Microsomal fatty acid elongation has been shown in hog aorta. The activity catalyzing the formation of fatty acids from acetyl and malonyl CoA was associated with a high molecular weight complex in the 6 x 10(6) g x min supernatant fraction. The principal product was palmitic acid. Some myristic and stearic acids were also formed. One elongation system was associated with protein which sedimented between 4500 g x min and 150,000 g x min. It used acetyl CoA but not malonyl CoA, and NADH was the preferred reducing agent. Radioactivity from acetyl CoA was incorporated into many fatty acids. In hog aorta a second elongation system was found associated with protein which sedimented at 6 x 10(6) g x min. It used malonyl CoA preferentially as substrate and either NADH or NADPH as reducing agent.

Impacts of water management options on flows in the Condamine River in Southern Queensland.

This paper examines the implications for river flows of a number of water practices and potential management options in the alluvial plains of the Upper Condamine River. It is an intensively cultivated area where irrigation is limited by the availability of water resources. The practice of capturing overland flows was investigated by the development of a model that simulates the performance of clusters of offstream storages up to sub-catchment scale. Management options examined included improvement to on-farm water use efficiency, the suppression of evaporation from open water storages, increasing the depth of those storages, decreasing their number, and improved tailwater return from irrigated land. Impacts of management options were analysed using a catchment scale water allocation model.