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PubMed1989

Genetic mapping in Xenopus laevis: eight linkage groups established.

Inheritance of alleles at 29 electrophoretically detected protein loci and one pigment locus (albinism) was analyzed in Xenopus laevis by backcrossing multiply heterozygous individuals generated by intersubspecies hybridization. Pairwise linkage tests revealed eight classical linkage groups. These groups have been provisionally numbered from 1 to 8 in an arbitrarily chosen order. Linkage group 1 includes ALB-2 (albumin), ADH-1 (alcohol dehydrogenase), NP (nucleoside phosphorylase), and ap (periodic albinism). Linkage group 2 contains ALB-1 and ADH-2, and probably is homeologous to group 1. Linkage group 3 comprises PEP-B (peptidase B), MPI-1 (mannosephosphate isomerase), SORD (sorbitol dehydrogenase), and mIDH-2 (mitochondrial isocitrate dehydrogenase). Linkage group 4 contains GPI-1 (glucosephosphate isomerase) and EST-4 (esterase 4). Linkage group 5 contains GPI-2 and PEP-D (peptidase D). Linkage group 6 comprises ACP-3 (acid phosphatase), sME (cytosolic malic enzyme), and GLO-2 (glyoxalase). Linkage group 7 consists of sSOD-1 (cytosolic superoxide dismutase), GPD-2 (glycerol-3-phosphate dehydrogenase), mME (mitochondrial malic enzyme), and the sex determining locus. Linkage group 8 includes FH (fumarate hydratase) and TRF (transferrin). Recombination frequencies between linked loci showed differences related to the genomic constitution (parental subspecies) and to the sex of the heterozygous parent. Independent assortment was observed between the duplicate ALB loci. This is true for the duplicate ADH, GLO, and MPI loci as well, supporting the view that these genes have been duplicated as part of a genome duplication that occurred in the evolutionary history of X. laevis. Comparative analysis of genetic maps reveals a possible conservation of several linkages from the Xenopus genome to the human genome.

Albinism↗
PubMed1979

Selenium, vitamin E and the response to swimming stress in the rat.

Experiments were conducted to determine the effects of exercise on rat glutathione peroxidase system enzymes and lipid peroxidation among animals supplemented and unsupplemented with selenium (Se) and vitamin E (E). Liver, muscle and blood were taken before, immediately after and 24 hours after exercising to exhaustion by swimming. No effect of exercise was found on muscle or liver enzymes, although exercise resulted in depressed glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G6PD) activities in erythrocytes immediately after exercise. Dietary Se supplementation did result in increased hepatic muscle and erythrocyte glutathione peroxidase activity, and decreased hepatic GR, G6PD and "malic enzyme" activities. Thiobarbituric acid reactive substances, and indicator of lipid peroxidation, increased in liver and muscle subsequent to exercise. This increase was reduced in liver, but not eliminated, by dietary E supplementation. The increase was not affected by dietary E in muscle, nor by dietary Se in either tissue.

Animals↗
PubMed1990

Dietary crude protein levels and the effect of isoproterenol on in vitro lipogenesis in the chicken.

Experiments were conducted with male broiler breeder chickens to determine the metabolic effects of two levels of dietary crude protein fed during a protein-reversal regimen on subsequent responses to a beta-adrenergic agonist (isoproterenol) in vitro. Day-old chickens were fed diets containing either 12 or 20% crude protein until 14 d of age and then switched to the opposite diet until 28 d. In experiment 1, chickens were killed at 28 d, and in experiment 2, at intervals following the switch at 14 d. In vitro lipogenesis and lipolysis were determined. The hepatic enzymes isocitrate dehydrogenase, malic enzyme and fatty acid synthetase were determined during the 14 to 28 d of age growth period. Both in vitro lipogenesis and lipolysis were greater at 28 d when chickens were fed the 12% rather than the 20% protein diet. Neither 0 to 14 d of age dietary treatment influenced 28-d metabolic parameters. In contrast, the 12% protein diet during the first 14-d period depressed body weight (160 g vs. 320 g; p less than 0.05) and weight gain during the 14 to 28 d of age period regardless of dietary crude protein levels (508 g vs. 626 g; p less than 0.05). Lipogenesis and lipolysis were at their highest points 4 d following the dietary switch. Isoproterenol decreased in vitro lipogenesis 70% in all dietary treatment groups. Stimulated lipolysis was not affected by nutritional status and was twice the unstimulated rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed2006

Protective effects of organic acids on survival of Escherichia coli O157:H7 in acidic environments.

Outbreaks of disease due to acid-tolerant bacterial pathogens in apple cider and orange juice have raised questions about the safety of acidified foods. Using gluconic acid as a noninhibitory low-pH buffer, we investigated the killing of Escherichia coli O157:H7 strains in the presence or absence of selected organic acids (pH of 3.2), with ionic strength adjusted to 0.60 to 0.68. During a 6-h exposure period in buffered solution (pH 3.2), we found that a population of acid-adapted E. coli O157:H7 strains was reduced by 4 log cycles in the absence of added organic acids. Surprisingly, reduced lethality for E. coli O157:H7 was observed when low concentrations (5 mM) of fully protonated acetic, malic, or l-lactic acid were added. Only a 2- to 3-log reduction in cell counts was observed, instead of the 4-log reduction attributed to pH effects in the buffered solution. Higher concentrations of these acids at the same pH aided in the killing of the E. coli cells, resulting in a 6-log or greater reduction in cell numbers. No protective effect was observed when citric acid was added to the E. coli cells. d-Lactic acid had a greater protective effect than other acids at concentrations of 1 to 20 mM. Less than a 1-log decrease in cell numbers occurred during the 6-h exposure to pH 3.2. To our knowledge, this is the first report of the protective effect of organic acids on the survival of E. coli O15:H7 under low-pH conditions.

Acetates↗
PubMed1977

Specificity of the disaccharide effect in the rat.

Male rats were starved and refed diets containing 40% disaccharides (maltose, trehalose, sucrose, turanose), trisaccharide (melezitose), starch, or the monosaccharide equivalents. Responses of hepatic glucose-6-phosphate dehydrogenase (G6PD), malic enzyme (ME), fatty acid synthetase (FAS), and total liver lipid (TLL) or changes in concentration of portal blood total carbohydrate and fructose were determined following refeeding of the different carbohydrate diets. Maltose, trehalose, sucrose, and turanose refeeding resulted in G6PD and ME responses which were higher than the responses to their component monosaccharides (disaccharide effect). Starch refeeding decreased the responses of G6PD, ME, and FAS when compared to refeeding of glucose. Refeeding diets containing fructose (sucrose, turanose, melezitose, and monosaccharide equivalents containing fructose) increased the responses of G6PD, ME, FAS, and TLL. No correlation between portal blood carbohydrate concentration and hepatic enzyme levels could be demonstrated. It is concluded that readily digestible disaccharides produce an effect which is greater than the effect produced by their monosaccharide equivalents. If these disaccharide configurations are fed as part of a trisaccharide or polysaccharide, the disaccharide effect is no longer discernible.

Animals↗
PubMed1977

Effect of premature weaning to different diets on the subsequent response to a dietary change.

Rats were weaned prematurely (day 18 postnatally) to a high-fat, high-carbohydrate or Purina Chow diet until aged 30 days. They were then fed the Purina Chow diet until aged 43 or 68 days. For a further 2 days they then received a high-fat diet. They were compared with rats weaned on day 30 to the Purina Chow diet. Adipose tissue of rats weaned prematurely to the high-carbohydrate diet or Purina Chow showed higher activities of phosphoenolpyruvate carboxykinase, malic enzyme and fatty acid synthetase than the same tissue from normally weaned rats or rats fed the high-fat diet from 18 to day 30. Prematurely weaned rats also have a higher level of blood cholesterol on day 45 after 2 days on the high-fat diet.

Adipose Tissue↗
PubMed2004

Effects of dihydrotestosterone on adipose tissue measured by serial analysis of gene expression.

Intra-abdominal fat accumulation is related to several diseases, especially diabetes and heart disease. Molecular mechanisms associated with this independent risk factor are not well established. Through the serial analysis of gene expression (SAGE) strategy, we have studied the transcriptomic effects of castration and dihydrotestosterone (DHT) in retroperitoneal adipose tissue of C57BL6 male mice. Approximately 50,000 SAGE tags were isolated in intact and gonadectomized mice, as well as 3 and 24 h after DHT administration. Transcripts involved in energy metabolism, such as glyceraldehyde-3-phosphate dehydrogenase, malic enzyme supernatant, fatty acid synthase, lipoprotein lipase, hormone-sensitive lipase and monoglyceride lipase, were upregulated by DHT. Transcripts involved in adipogenesis, and cell cycle and cell shape organization, such as DDX5, C/EBPalpha, cyclin I, procollagen types I, III, IV, V and VI, SPARC and matrix metalloproteinase 2, were upregulated by DHT. Cell defense, division and signaling, protein expression and many novel transcripts were regulated by castration and DHT. The present results provide global genomic evidence for a stimulation of glycolysis, fatty acids and triacylglycerol production, lipolysis and cell shape reorganization, as well as cell proliferation and differentiation, by DHT. The novel transcripts regulated by DHT may contribute to identify new mechanisms involved in the action of sex hormones and their potential role in obesity.

Adipose Tissue↗
PubMed2004

Effects of ketanserin on hypergranulation tissue formation, infection, and healing of equine lower limb wounds.

The objective of this multicentre, randomized, controlled field study was to determine the efficacy of ketanserin gel in preventing exuberant granulation tissue formation (hypergranulation) and infection in equine lower limb wounds. Horses and ponies (n = 481) with naturally occurring wounds were randomized to either topical treatment with ketanserin gel (n = 242) or a positive control (Belgium, Germany: ethacridin lactate solution, n = 120; France, United Kingdom: malic, benzoic, and salicylic acid [MBS] cream, n = 119). Treatment continued until the wound healed (success), formed hypergranulation tissue (failure), or became infected (failure). Treatment was terminated after 6 months in all remaining animals. Ketanserin was successful in 88% of cases. Wounds treated with ketanserin were 2 and 5 times more likely to heal successfully than were those treated with MBS or ethacridin lactate, respectively. Ketanserin gel is thus more effective than these standard treatments in preventing hypergranulation tissue and infection of equine lower limb wounds.

Administration, Topical↗
PubMed1992

Interaction of androgens and prolactin on prostatic enzymes of the pyruvate-malate cycle involved in lipogenesis in castrated mature monkey, Macaca radiata.

Effects of androgens, prolactin (Prl) and bromocriptine (Br) on the specific activities of prostatic (caudal and cranial) enzymes of the pyruvate-malate cycle were studied in castrated mature bonnet monkeys. Castration decreased the activity of NADP+ isocitrate dehydrogenase (ICDH), ATP citrate lyase, malate dehydrogenase (MDH), malic enzyme and fatty acid synthase (FAS). Administration of testosterone propionate (TP)/dihydrotestosterone (DHT) increased the activities of all these enzymes in both lobes. Malate dehydrogenase maintained normal activity. Prl also had a stimulatory effect on the enzymes and was further enhanced when Prl was given in combination with TP/DHT. Unlike Prl, bromocriptine treatment inhibited all the enzymes in both lobes. Thus, prolactin was found to have a direct as well as a synergistic effect with androgens on enzymes of the pyruvate-malate cycle in the prostate of castrated mature monkeys.

ATP Citrate (pro-S)-Lyase↗
PubMed1986

A new povidone-iodine cream for the treatment of burns. Comparison with a standard topical regimen.

A remarkable improvement in the rate of burn healing has been achieved with a mixture of povidone-iodine ointment (Betadine) and malic, benzoic and salicylic acids (MBS) (Aserbine). A study was undertaken to compare the effects of a new povidone-iodine formulation (Betadine cream) with and without MBS with povidone-iodine ointment plus MBS. All preparations were easy to apply and were readily removed, causing only mild discomfort on application in the majority of cases. A significant difference in healing times was observed between povidone-iodine cream and povidone-iodine cream plus MBS. There was also a significant difference in the decrease in the number of positive bacterial cultures between these two treatments. This applied to both superficial and deep burns. No skin sensitivity reactions were reported with any of the preparations. The addition of MBS to povidone-iodine cream did not produce as significant an improvement in results as its addition to povidone-iodine ointment.

Administration, Topical↗
PubMed1982

[Mechanism of the Crabtree effect: influence of pH on the rate of mitochondrial respiration].

A detailed study of dependence of mitochondrial respiration on pH of incubation medium has been carried out. It was shown that during oxidation of malic, succinic and ascorbic acids mitochondrial respiration shows a sharp fall, when the pH of incubation medium is decreased from 7.5 to 6.5. It was assumed that one of the possible causes of the Crabtree effect is a reduction of the pH value of the cytoplasm.

Alcohol Oxidoreductases↗
PubMed1995

Increased urinary excretion of analogs of Krebs cycle metabolites and arabinose in two brothers with autistic features.

A marked increase in analogs of Krebs cycle metabolites was found in the urine of two brothers with autistic features. These metabolites included citramalic, tartaric (3-OH-malic), and 3-oxoglutaric acids and compounds tentatively identified as a citric acid analog and partially identified as a phenylcarboxylic acid by the fragmentation pattern of the trimethylsilyl (TMS) derivatives of the compounds and mass shifts of the same compounds derivatized with perdeuterated N,O-bis(trimethylsilyl)trifluoroacetamide. The molecular mass of the TMS derivative of the tentatively identified citric acid analog was 596 Da, based on a finding of a significant M - 15 ion at m/z 581. The citric acid analog was excreted in quantities as high as 137 mmol/mol creatinine, based on the response factor of citric acid as a surrogate calibrator. A carbohydrate with a retention time and mass spectrum identical to arabinose was also found in high concentrations in the urine of these brothers.

Arabinose↗
PubMed1994

Molecular analysis of the malic enzyme gene (mae2) of Schizosaccharomyces pombe.

Sequence analysis of a 4.6-kb HindIII fragment containing the malic enzyme gene (mae2) of Schizosaccharomyces pombe, revealed the presence of an open reading frame of 1695 nucleotides, coding for a 565 amino acid polypeptide. The mae2 gene is expressed constitutively and encodes a single mRNA transcript of 2.0 kb. The mae2 gene was mapped on chromosome III by chromoblotting. The coding region and inferred amino acid sequence showed significant homology with 12 malic enzyme genes and proteins from widely different origins. Eight highly homologous regions were found in these malic enzymes, suggesting that they contain functionally conserved amino acid sequences that are indispensable for activity of malic enzymes. Two of these regions have previously been reported to be NAD- and NADP-binding sites.

Amino Acid Sequence↗
PubMed1990

Potential toxicity of presumably insoluble aluminum salts in presence of common dietary acids.

It has recently been shown that aluminum absorption may occur following the administration of oral aluminium-containing phosphate-binders and antacids. Computer simulations based on relevant aluminium complex equilibria have been used in the present work to investigate the potential influence of the simultaneous ingestion of common dietary acids on this phenomenon. It results from these studies that aluminium absorption may be favoured to various extents in the presence of citric, malic, oxalic, succinic and tartric acids.

Administration, Oral↗
PubMed1996

Diet fat alters expression of genes for enzymes of lipogenesis in lean and obese mice.

The objective of this experiment was to determine the effect of polyunsaturated fatty acids on gene expression for fatty acid synthase, acetyl CoA-carboxylase, malic enzyme, pyruvate kinase, and phosphoenolpyruvate carboxykinase in obese mice. Eight-week-old female lean and obese mice were fed semi-purified diets containing 20% (w/w) fat of either high or low polyunsaturated to saturated (P/S) fatty acid ratio for four weeks. Total RNA was isolated from liver and was hybridized to cDNA probes for the above enzymes. Consumption of a high P/S diet decreased mRNA levels for all the lipogenic enzymes studied in both lean and obese mice. Compared to lean mice, obese mice exhibited a higher mRNA level for fatty acid synthase, acetyl CoA-carboxylase, malic enzyme, and pyruvate kinase in animals fed either a high or low P/S diet. Enzyme-specific activities followed the same profile as the mRNA levels in both lean and obese mice fed a high or low P/S diet. The decrease in liver fatty acid synthase mRNA level was more pronounced in lean mice compared to obese mice, suggesting that the obese mice may be more resistant to polyunsaturated fatty acid feedback control of gene expression.

Acetyl-CoA Carboxylase↗
PubMed1999

Effect of dietary fat on whole body fatty acid synthesis in weanling rats.

The effect of dietary fat on body composition, whole body lipogenesis, and enzyme activity was measured in rats over the first 16 weeks post-weaning. Rats were fed either a low fat (5% w/w fat) or high fat (20% w/w fat) diet for the first 4 weeks. After this time all rats were fed the low fat diet. The results showed no significant effect of diet on the rate of fat synthesis over the first 8 weeks of the experiment. However, the activities of the enzymes of fatty acid synthesis [glucose 6-phosphate dehydrogenase, malic enzyme, adenosine triphosphate-citrate lyase, acetyl-coenzyme A carboxylase (ACCX), fatty acid synthetase] were dependent on the age and dietary status of the animals. The exact pattern depended on the specific enzyme and the tissue source. No significant differences in pyruvate dehydrogenase (PDH) activity were observed. Mathematical analysis of the enzyme activities suggested that ACCX and PDH were the most likely sites of fat synthesis regulation. In addition, an examination of body composition and overall weight retention showed that the "weight increasing" effect of a high fat diet could be completely reversed by subsequent feeding of a low fat diet. However, the reversal required an additional 12 weeks. Interestingly, at this time the rats switched from a high fat to a low fat diet had a lower body weight and lower body fat content than rats fed a low fat diet throughout the course of the experiment.

Journal Article↗
PubMed2000

Application of Convective Interaction Media (CIM) disk monolithic columns for fast separation and monitoring of organic acids.

The separation of organic acids on the anion-exchange monolithic support, commercially available as Convective Interaction Media (CIM), is presented in this study. It is demonstrated that citric, isocitric, pyruvic, fumaric, malic, and alpha-ketoglutaric acid can be successfully separated using a CIM monolithic column of suitable user-adjustable length. The effect of the mobile phase composition on the separation is investigated. CIM monolithic columns of adjustable length from 3 to 18 mm are compared regarding the resolution and the back pressure. It is shown that the CIM monolithic column of 12 mm in length enables a good separation of all six organic acids within 3 min and exhibits a linear dependence of back pressure versus flow rate. The resolution and the dynamic binding capacity are found to be flow-unaffected. A filtrated sample of bioprocess supernatant is analyzed without previous pretreatment, which indicates the possibility of online monitoring of small molecules during the bioprocess using CIM monolithic columns.

Journal Article↗