Biomedical subjects
Publications and source records attributed to S Reiser.
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To provide 4-hydroxybutyryl-CoA for poly(3-hydroxybutyrate-co-4-hydroxybutyrate) formation from glutamate in Escherichia coli, an acetyl-CoA:4-hydroxybutyrate CoA transferase from Clostridium kluyveri, a 4-hydroxybutyrate dehydrogenase from Ralstonia eutropha, a gamma-aminobutyrate:2-ketoglutarate transaminase from Escherichia coli, and glutamate decarboxylases from Arabidopsis thaliana or E. coli were cloned and functionality tested by expression of single genes in E. coli to verify enzymatic activity, and uniquely assembled as operons under the control of the lac promoter. These operons were independently transformed into E. coli CT101 harboring the runaway replication vector pJM9238 for polyhydroxyalkanoate (PHA) production. Plasmid pJM9238 contains the PHA biosynthetic operon of R. eutropha under tac promoter control. Polyhydroxyalkanoate formation was monitored by nuclear magnetic resonance (NMR) spectroscopic analysis of the chloroform extracted and ethanol precipitated polyesters. Functionality of the biosynthetic pathway for copolymer production was demonstrated through feeding experiments using various carbon sources that supplied different precursors within the 4HB-CoA biosynthetic pathway.
Acinetobacter calcoaceticus BD413 accumulates wax esters and triacylglycerol under conditions of mineral nutrient limitation. Nitrosoguanidine-induced mutants of strain BD413 were isolated that failed to accumulate wax esters under nitrogen-limited growth conditions. One of the mutants, Wow15 (without wax), accumulated wax when grown in the presence of cis-11-hexadecenal and hexadecanol but not hexadecane or hexadecanoic acid. This suggested that the mutation may have inactivated a gene encoding either an acyl-acyl carrier protein or acyl-coenzyme A (CoA) reductase. The Wow15 mutant was complemented with a cosmid genomic library prepared from wild-type A. calcoaceticus BD413. The complementary region was localized to a single gene (acr1) encoding a protein of 32,468 Da that is 44% identical over a region of 264 amino acids to a product of unknown function encoded by an open reading frame associated with mycolic acid synthesis in Mycobacterium tuberculosis H37Ra. Extracts of Escherichia coli cells expressing the acr1 gene catalyzed the reduction of acyl-CoA to the corresponding fatty aldehyde, indicating that the gene encodes a novel fatty acyl-CoA reductase.
1. To compare the impact of type of carbohydrate, genotype and phenotype on the synthesis and levels of plasma lipoprotein protein. Sprague-Dawley rats and carbohydrate-sensitive LA/N-corpulent (cp) rats were fasted (2 days) and then fed diets containing 54% carbohydrate as either sucrose, fructose or cooked cornstarch for 2 days. 2. The amount of 3H-protein present in the VLDL + chylomicron fraction of Sprague-Dawley rats 2 hr after injection of 3H-leucine was affected by type of dietary carbohydrate: sucrose greater than fructose greater than starch. 3. Obese and lean LA/N-cp rats fed diets containing sucrose or fructose had lower concentrations of HDL protein and higher levels of 3H-protein in VLDL + chylomicron fraction than those fed starch. 4. Obese LA/N-cp rats had more HDL protein and higher levels of 3H-protein in VLDL + chylomicron fraction than their lean littermates.
Two hundred sixty-six free-living human volunteers, 21-64 y old, were trained by dietitians to record daily their food intake for at least 7 d. Subsequently, they were fed diets of conventional foods adjusted in amounts to maintain their body weight for greater than or equal to 45 d. Comparing their estimated energy intake with the intake determined to maintain weight yielded mean differences of 2365 and 1792 kJ (565 and 428 kcal) in men and women, respectively, representing an underreporting of 18%. Twenty-two individuals (8%) overestimated and 29 (11%) were accurate to within 419 kJ (100 kcal) of their maintenance requirement. The remaining 215 individuals (81%) reported their habitual intake at 2930 +/- 1586 kJ (700 +/- 379 kcal) below that subsequently determined as their maintenance requirement. These findings suggest caution in the interpretation of food-consumption data.
Inadequate dietary copper is known to result in undesirable metabolic changes in rats and humans. Abnormal cardiac function, leading to sudden death, is a common finding when copper deficient rats are fed a 62% fructose diet. To further study the apparent mineral-carbohydrate relationship to cardiac physiology, 3 male and 3 female swine were randomly assigned to four groups (6 pigs per group) which were fed low copper (1.5 ppm) or copper supplemented (40 ppm) diets with 20% of calories from either fructose or glucose for 10 weeks. In agreement with results from other animal studies, copper deficient swine exhibited decreased plasma ceruloplasmin, erythrocyte superoxide dismutase and plasma lysyl oxidase activities and lowered serum copper. The copper deficient fructose group had the lowest aortic lysyl oxidase activity and hematocrit when compared to the other groups. The relative heart weight in the copper deficient fructose group was 93% greater than the other three dietary groups. The livers of copper deficient fructose fed pigs were also significantly larger. Two enzymes related to cardiac and hepatic function, aspartate and alanine aminotransferase were also measured. Copper deficiency significantly lowered alanine aminotransferase but there was no dietary effect on aspartate amino-transferase. The results of this project indicate that the pig is a sensitive model for the study of cardiovascular abnormalities which occur when fructose is consumed with a low copper diet.
Eleven male and nine female adult subjects were given one of the following five carbohydrate-drink combinations (per kg body wt) on five mornings separated by greater than or equal to 2 wk: 1) 1.0 g glucose, 2) 0.9 g uncooked cornstarch, 3) 1.0 g glucose followed 20 min later by 1.75 g fructose, 4) 0.9 g uncooked cornstarch followed 20 min later by 1.75 g fructose, and 5) water followed 20 min later by 1.75 g fructose. Glucose plus fructose was the most insulinogenic followed by glucose alone, starch plus fructose, starch alone, and water plus fructose. The urinary losses of chromium followed a similar pattern. Subjects with the highest concentrations of circulating insulin displayed decreased ability to mobilize chromium on the basis of urinary chromium excretion. Therefore, urinary chromium losses are related to the insulinogenic properties of carbohydrates.
Ten hyperinsulinemic and 11 nonhyperinsulinemic men consumed for 5 wk each in a cross-over design a diet, similar to one currently consumed in the United States, with 20% of the kilocalories from either fructose or high-amylose cornstarch to determine the effects of the two diets on various blood metabolites considered to be risk factors associated with heart disease. In the hyperinsulinemic men the intake of fructose as compared with cornstarch significantly increased total triglycerides and their lipoprotein distribution; total and very-low-density lipoprotein cholesterol; apoproteins B-100, C-II, C-III; and uric acid. In the nonhyperinsulinemic men total triglycerides, total and low-density lipoprotein cholesterol and uric acid were significantly greater after the consumption of fructose than after cornstarch. The results indicate that in a diet high in saturated fatty acids and cholesterol, fructose increases the levels of risk factors associated with heart disease, especially in hyperinsulinemic men.
A balance study was conducted to assess the effects of consuming low-copper diets, high in fructose or cornstarch. The study involved 19 apparently healthy males, aged 21-57 y. The two experimental diets averaged 0.35 mg Cu/1000 kcal and provided 20% of the calories from fructose or cornstarch. Cu, zinc, calcium, magnesium, and iron balances were determined 1 wk before the study (pretest) when the subjects consumed self-selected diets and after consuming the experimental diets for 6 wk. No major differences in mineral balances were evident between the two groups during the pretest study when the subjects ate self-selected diets. In contrast, when fed the test diets, the group consuming the low-Cu fructose diet had significantly more positive balances for all minerals studied than the group fed the low-Cu cornstarch diet. The results indicate that dietary fructose enhances mineral balance.
Ten hyperinsulinemic and 11 nonhyperinsulinemic men consumed a typical American diet containing 20% of calories either as fructose or as high-amylose cornstarch for 5 wk in a crossover design to determine their effects on indices of glucose tolerance. Blood glucose, insulin, and fructose were determined before and 30, 60, 90, 120 and 180 min after breakfast, lunch, and dinner. Glucose responses were significantly lower 60 and 120 min and the insulin response lower 60 min after the meals containing fructose. Hyperinsulinemic men showed a tendency toward decreased insulin sensitivity after consuming fructose on the basis of an increased insulin-to-glucose ratio and decreased insulin binding to erythrocytes. These results generally show a beneficial effect of fructose on glucose tolerance; however, recommendations for including large amounts of fructose in the diet should also be based on a complete evaluation of the effect on other metabolic risk factors.
The purpose of this study was to determine whether the interaction between fructose and copper status can be affected by the mode of copper administration. Male Sprague-Dawley rats (n = 94) were fed from weaning diets that contained 0.65-0.85 mg copper/kg diet for 42 d. The experimental design of the study was a 2 x 2 x 3 analysis of variance: two types of carbohydrate (fructose, corn starch), two modes of copper delivery (orally or subcutaneously) and three different extrinsic copper levels (0, 1.5 or 3 mg/kg diet or 0, 11.25 or 22.5 micrograms Cu/d by injection). In the rats fed fructose, direct indicators of copper status such as plasma copper and tissue superoxide dismutase activity were increased to a much greater extent when copper was supplied subcutaneously rather than orally as part of the diet. In the starch-fed rats the effect of the mode of copper delivery on the levels of these indices was usually not significantly different. As a result, the feeding of fructose as compared to starch produced lower levels of direct indicators of copper status when copper was supplied in the diet but not subcutaneously. It appears that ingestion of fructose as compared to starch increases the requirement for copper and that the mode of copper administration that circumvents the gastrointestinal tract attenuates the fructose-copper interaction. Thus, the fructose-copper interaction occurs either during intestinal digestion and absorption, hepatic uptake of copper via the portal blood or its hepatic utilization.
The effects of feeding diets containing fructose, glucose or starch with or without added magnesium (Mg) on tissue levels of calcium (Ca) in weanling rats were investigated. Rats were fed their respective diet for 4 wk and then fasted overnight. After decapitation, blood was immediately collected, and liver, kidney and heart were removed to determine their Mg and Ca content. Relative kidney and liver weights were greatest in the rats fed the fructose diet. Hypomagnesemia was observed in all animals fed a Mg-deficient diet regardless of carbohydrate source (1.71 vs. 2.27 mg Mg/dl plasma for Mg-deficient vs. Mg-adequate groups, respectively). Among Mg-deficient groups, starch-fed rats had the highest level of plasma Mg, while fructose-fed rats had the highest level of kidney Mg. Although plasma and liver Ca levels were not significantly different among the groups, Ca content of the kidney and heart was greatest in animals fed a Mg-deficient diet with fructose. This was especially evident in the Ca content of the kidney, which was 8- to 9-fold greater than that of all other experimental groups. The results of the present study demonstrate the presence of a synergistic interaction between fructose and Mg deficiency that increases Ca levels in certain soft tissues, especially the kidney.
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Copper deficiency has been shown to result in severe cardiovascular lesions in several species of animals. The principal carbohydrate in the copper-deficient diet most often used with rats is sucrose, which is known to have adverse effects on carbohydrate and lipid metabolism and thus may contribute to cardiovascular disorders. These observations prompted experiments in which starch and fructose were substituted for sucrose in a copper-deficient diet, to see if the effects of the copper deficiency might be modified. In the hearts from rats fed copper-deficient diets with fructose or sucrose, there was marked, mostly ventricular hypertrophy, and mild to severe myocardial inflammation, degeneration, and fibrosis. Aneurysm of the left ventricle and pericarditis also were common. Hearts from the starch, copper-deficient groups were much less hypertrophic, and very few were affected by myocardial inflammation, degeneration, or fibrosis. Defects of elastin or other structures were not observed in the aortas or pulmonary or coronary arteries of any specimens.
This study describes the progressive changes in exocrine pancreatic enzyme specific activities from rats provided with a diet containing either 62% starch (S) or fructose (F) with 6.0 ppm copper (+Cu) or 0.6 ppm (-Cu) from 21 to 61 days of age. Copper-deficient rats had significantly reduced pancreatic weight and DNA, RNA, protein and copper content and superoxide dismutase (SOD) activity after consuming diet for 39 days. These reductions were far more severe in F-Cu than in S-Cu rats. Copper deficiency significantly reduced pancreatic amylase and lipase specific activities, but the effect was more severe in the F-Cu dietary group. Chymotrypsinogen and trypsinogen specific activities were reduced only in the F-Cu dietary group and the CHO x Cu interaction was significant. High dietary fructose and low dietary copper in combination interact to produce severe reductions in pancreatic amylase, lipase, chymotrypsinogen, and trypsinogen specific activities which could not be explained by either copper deficiency or by an increase in dietary fructose alone.
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Twenty male subjects, 11 normal and 9 carbohydrate-sensitive, participated in a study in which the effect of feeding diets low in copper (1.03 mg/d) on the number and affinity of insulin receptors was determined. Since carbohydrate-sensitive subjects are hyperinsulinaemic, it was anticipated that they would demonstrate a down-regulation of insulin receptors. The subjects were fed a low copper diet for 11 weeks and then replenished with copper (3 mg/d) for 3 weeks. Regardless of diets fed, carbohydrate-sensitive subjects showed increased insulin binding. The increase was due to the number of receptors without any change in their affinity. This unusual and unexpected observation in carbohydrate-sensitive subjects suggests an altered response of the insulin receptor, namely the failure of plasma insulin to down-regulate the number of receptors. It is possible that such an alteration could lead to a shift of the biological response curve of insulin to the left in some target tissues thereby causing hyperlipaemia by diverting glucose into triglyceride. The exact mechanism of hyperlipaemia in carbohydrate-sensitive subjects remains to be clarified.
For 13 weeks, 20 men (aged 23 to 56), 19 premenopausal (aged 21 to 48), and 14 postmenopausal women (aged 49 to 65) consumed a 7-day rotation menu conforming to dietary recommendations of several research and health organizations. The diets were designed to make minimal changes in the standard American diet by the use of well-accepted normal foods. The average daily composition of the diet was 50% carbohydrate (complex 35%, simple 15%), 35% fat (P:S 0.7), and 15% protein; it contained 100 mg cholesterol, 1 gm sodium, and 14.5 gm neutral detergent fiber per 1,000 kcal. The acceptability of the menus was examined through questionnaires administered after the experimental diet period. Subjects rated the menus and individual foods and made recommendations for improvement of the menus. Most subjects (33 of 41 responding) rated the menu better than or almost as good as their usual diet. Few (9) were ever hungry during the 13-week period. During the study, a number of biochemical parameters were measured; the measurements indicated beneficial results. Blood pressure, cholesterol levels, and glucose responses were all improved after consumption of the menu. The results indicate that with minimal changes in the normal U.S. diet, acceptable menus that have beneficial effects on risk factors for heart disease and diabetes can be developed.