Response to stress by riboflavin-deficient and pantothenic acid-deficient dogs.
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To date, the pathogenesis of systemic lupus erythematosus (SLE) remains unclear. By critically analyzing clinical facts and laboratory data, a hypothesis is proposed: drug-induced lupus erythematosus (DILE) is linked to a deficiency in Coenzyme A (CoA) that is secondary to a deficiency in pantothenic acid. This hypothesis is used to explain the high incidence of SLE in females, the role of sex hormones in this disease and the mechanism underlying a flare. The actions of anti-malarials and steroids are also discussed. The protean clinical presentation of SLE is attributed to co-existing deficiencies of dietary factors in addition to pantothenic acid. Contributing factors to these deficiencies may include increased nutritional requirements resulting from gene mutations. Treatment is replacement therapy with doses of pantothenic acid that is hundreds of times higher than that of the Dietary Reference Intake (DRI) and other vitamins. Using this method, 12 SLE females were studied with promising results.
C1-THF (5,6,7,8-tetrahydrofolate) synthase is a trifunctional protein catalyzing the sequential reactions specified by the enzymes 10-formyl-THF synthetase (EC 6.3.4.3), 5,10-methenyl-THF cyclohydrolase (EC 3.5.4.9), and 5,10-methylene-THF dehydrogenase (EC 1.5.1.5). These three activities supply the activated one-carbon units required for the biosynthesis of purines, thymidylate, the amino acids histidine and methionine, the vitamin pantothenic acid, and the formyl group of mitochondrial fMet-tRNAfMet. Extracts of Saccharomyces cerevisiae whose growth is dependent on the three activities of C1-THF synthase contain 2-3 times the level of enzyme activity of extracts from cells grown under conditions where they are independent of this enzyme. Repression of C1-THF synthase activity requires the simultaneous presence of adenine, histidine, methionine, and pantothenic acid. Starvation of the cells for any one of these nutrients leads to derepression of the enzyme. Drug-induced folate starvation also leads to derepression of enzyme activity. The response to changing nutritional conditions occurs within 1 h and is due to changes in the steady-state concentration of C1-THF synthase enzyme, rather than to activation or deactivation of a pre-existing pool of enzyme. Determination of the amount of C1-THF synthase mRNA under the various growth conditions by an in vitro translation/immunoprecipitation assay indicates that regulation of the enzyme occurs predominantly at a pretranslational level since steady-state levels of C1-THF synthase mRNA are 2-3-fold higher in derepressed cells than in repressed cells.
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We investigated the levels of water-soluble vitamins except for vitamin B6 in the blood and urine of Japanese college male (n = 10) and female (n = 10) students. They consumed for 7 d a semi-purified diet based on Japanese Dietary Reference Intakes to assess the Recommended Dietary Allowances (RDA) for water-soluble vitamins and to present some new normal values for blood and urine levels of water-soluble vitamins in Japanese. The blood and the 24-h urine samples were collected on the last day of the experiment and measured. The values of total vitamin B1 in whole blood, total vitamin B2 in whole blood, total cyanocobalamin in serum, total nicotinamide in whole blood, total pantothenic acid in whole blood, total folates in serum, total biotin in serum, and ascorbic acid in plasma were 104+/-17 pmol/mL (mean+/-SD), 216+/-25 pmol/mL, 0.34+/-0.05 pmol/mL, 59.1+/- 5.0 nmol/ mL, 2.45+/-0.37 nmol/mL, 15.6+/-4.6 pmol/mL, 8.3+/-0.5 pmol/mL, and 62+/-10 nmol/mL, respectively, in males, and 90+/-23, 234+/-18, 0.67+/-0.20, 61.9+/-6.0, 2.48+/-0.30, 30.2+/-8.6, 8.4+/-0.3, and 67+/-14, respectively, in females. There was a significant difference in the values of cyanocobalamin and total folates between men and women. The urinary excretion of vitamin B1, vitamin B2, cyanocobalamin, sum of the catabolic metabolites of nicotinamide, pantothenic acid, folates, biotin, and ascorbic acid were 665+/-114 nmol/d, 562+/-325 nmol/d, 93+/-31 pmol/d, 84+/-26 micromol/d, 9.3+/-2.3 micromol/d, 19.4+/-2.8 nmol/d, 83+/-18 pmol/d, and 148+/-51 micromol/d, respectively, in males, and 495+/-212, 580+/-146, 145+/-49, 83+/-19, 16.9+/-1.3, 22.7+/-2.7, 83+/-23, and 140+/-51, respectively, in females. There was a significant difference in the urinary excretion of cyanocobalamin, pantothenic acid and total folates between men and women. These values will be useful for the nutritional assessment of water-soluble vitamins for Japanese, although the examination period was short. In future, an experiment with various age groups and re-evaluation by repeated experiments will provide more accurate values.
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Forty-eight growing pigs were randomly assigned to five dietary groups and penned individually. They received a diet based on barley, wheat, corn and soya bean meal according to requirement. The experimental groups were supplemented with 400% or 800% of vitamins B(2), B(6) and pantothenic acid, or 400% or 800% of biotin, while all other vitamins were administered according to requirement. Growth performance, carcass characteristics, aspartate aminotransferase (AST), and content of vitamins in blood, liver and muscles were recorded. Growth performance showed no influence of supplementation, while backfat thickness in the group with 800% B(2)/B(6)/pantothenic acid was significantly higher. Content of B(2) in blood, liver and muscle was similar in all groups. Content of B(6) in blood and liver showed significant differences according to supplementation. The content of vitamin B(6) in muscle in the experimental groups was significantly higher than that in the control group. The content of pantothenic acid in blood and muscle in the experimental groups was significantly higher, while in liver all groups were significantly influenced by the supplementation level. Biotin content in liver showed no influence, but the content in plasma was significantly higher in the experimental groups and the content in muscle was significantly higher according to supplementation. The activity of AST showed no significant influence of the dietary vitamin level, but it was obviously decreased in the groups supplemented with biotin. The findings indicate that the dietary supplementation of vitamin B(2), B(6), pantothenic acid and biotin could not improve performance, but the contents in blood, liver and muscle.
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The polyenzyme complex responsible for the synthesis of tyrocidine in Bacillus brevis (ATCC 8185) was found to contain 4'-phosphopantetheine, which appeared to be connected with the production of growing peptide chains. Confirmation of this assumption has now been obtained by purifying from bacterial lysates a polyenzyme-dissociation product; this was labeled with [(14)C]pantothenic acid and peptide chains containing tritiated amino acids, and had a molecular weight of 17,000. To obtain these results, organisms were grown udner conditions favorable for incorporation of radioactive pantothenic acid into tyrocidine-synthesizing enzymes. A crude lysate of the [(14)C]pantothenic acid-labeled organisms was preincubated with the tritiated amino acids to form enzyme-bound growing peptide chains. The doubly labeled fragments were purified from the polyenzyme-dissociation products produced by prolonged lysis. In a second set of experiments, the three enzymes responsible for tyrocidine synthesis, including the two polyenzymes containing pantetheine, were purified and incubated with radioactive amino acids and ATP to form polyenzyme-bound peptide chains. Thereupon, a Triton X-100 extract of the 20,000 x g fraction of crude homogenate was added to dissociate the purified polyenzymes. The dissociation products were purified and yielded, on dodecyl sulfate gel electrophoresis, peptidyl-marked products ranging in molecular weight from 90,000 to 17,000, the latter being most abundant. Electrophoresis of analogous preparations after preincubation with higher concentrations of dodecyl sulfate and dithiothreitol at 100 degrees yielded a single product of 17,000 molecular weight, indicating that the larger molecular weight fractions were aggregates thereof.
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Two Salmonella typhimurium strains, which could be used as sources for the leucine biosynthetic intermediates alpha- and beta-isopropylmalate were constructed by a series of P22-mediated transductions. One strain, JK527 [flr-19 leuA2010 Delta(leuD-ara)798 fol-162], accumulated and excreted alpha-isopropylmalate, whereas the second strain, JK553 (flr-19 leuA2010 leuB698), accumulated and excreted alpha- and beta-isopropylmalate. The yield of alpha-isopropylmalate isolated from the culture medium of JK527 was more than five times the amount obtained from a comparable volume of medium in which Neurospora crassa strain FLR(92)-1-216 (normally used as the source for alpha- and beta-isopropylmalate) was grown. Not only was the yield greater, but S. typhimurium strains are much easier to handle and grow to saturation much faster than N. crassa strains. The combination of the two regulatory mutations flr-19, which results in constitutive expression of the leucine operon, and leuA2010, which renders the first leucine-specific biosynthetic enzyme insensitive to feedback inhibition by leucine, generated limitations in the production of valine and pantothenic acid. The efficient, irreversible, and unregulated conversion of alpha-ketoisovaleric acid into alpha-isopropylmalate (alpha-isopropylmalate synthetase K(m) for alpha-ketoisovaleric acid, 6 x 10(-5) M) severely restricted the amount of alpha-ketoisovaleric acid available for conversion into valine and pantothenic acid (ketopantoate hydroxymethyltransferase K(m) for alpha-ketoisovaleric acid, 1.1 x 10(-3) M; transaminase B K(m) for alpha-ketoisovaleric acid, 2 x 10(-3) M).
Three proteins of the inner mitochondrial membrane of Neurospora crassa were found to be covalently modified with a derivative of pantothenic acid. One of these proteins is a subunit of cytochrome c oxidase and two are subunits of the ATPase-ATP synthase. Cells of a pantothenate auxotroph of N. crassa were labeled with [14C]pantothenic acid, and mitochondrial proteins containing radiolabeled pantothenate were detected by electrophoresis of detergent-solubilized mitochondria. Mitochondria from cells that were colabeled with [14C]pantothenate and [3H]leucine were reacted with specific antisera against the cytochrome c oxidase and F1-ATPase enzyme complexes. Electrophoresis of the labeled subunits of these isolated complexes showed that the [14C]pantothenate-associated peptides corresponded to [3H]leucine-labeled subunit 6 of cytochrome c oxidase and two [3H]leucine-labeled subunits (tentatively identified as subunits 8 and 11) of the ATPase-ATP synthase. Pantothenate modification of these enzyme subunits, which are synthesized on extramitochondrial ribosomes, may contribute to their transport and assembly into mitochondria, or it may participate in the catalytic activity of the assembled enzymes.
1. The amounts of magnesium, copper, zinc, phosphorus, manganese, potassium, vitamin B6, vitamin B12, folate, pantothenic acid, biotin, vitamin E and dietary fibre in the British household diet were calculated by applying appropriate values from recent analytical studies to the amounts of foods recorded in the National Food Survey during 1986. 2. National average intakes were (mg/person per d): Mg 247, Cu 1.25, Zn 9.0, P 1249, Mn 3.43, K 2694, vitamin B6 1.73, vitamin B12 6.33 micrograms, folate 230 micrograms, pantothenic acid 6.07, biotin 35 micrograms, vitamin E 8.4. Regional and income-group differences were estimated, and found to be small. Additional contributions from alcoholic drinks and confectionery were also determined. 3. Dietary fibre was estimated both as unavailable carbohydrate and as non-starch polysaccharide. The national average intakes were 21.8 and 12.9 g/d respectively. 4. Intakes were compared with Canadian (Department of National Health and Welfare, 1983) and American (National Research Council, 1980) recommended dietary allowances (RDAs). With the exception of biotin, the Canadian RDAs were met by the household diet but the much higher American RDAs were only met for vitamin B12 and pantothenic acid.
Ten sets of 5 littermate pigs from each of 2 genetic strains were utilized to determine the impact of the dietary concentration of 5 B vitamins (riboflavin, niacin, pantothenic acid, cobalamin, and folacin) on growth from 9 to 28 kg of BW in pigs with high or moderate capacity for lean growth. All pigs (penned individually) were reared via a segregated, early weaning scheme, so that the lean growth potential of each strain could be expressed. The basal diet provided the 5 test vitamins at concentrations of total and estimated bioavailability equivalent to a minimum of 100 and 70%, respectively, of their estimated requirements (NRC, 1998) for 5- to 10-kg pigs. At a BW of 9 +/- 0.9 kg, pigs within each litter were allotted to the basal diet supplemented with sources of the 5 test vitamins equivalent to an additional 0, 100, 200, 300, or 400% (bioavailable) of the NRC requirements. Pigs from the high lean strain consumed less feed (P < 0.05) and gained BW faster (P < 0.02) and more efficiently (P < 0.01) than pigs of the moderate lean strain. In both lean strains, the rate and efficiency of growth were improved (P < 0.01) as dietary B vitamin concentrations were increased. However, the dietary B vitamin concentrations needed to optimize G:F were greater (P < 0.03) in the high (>470% of NRC, 1998) vs. moderate (270%) lean strain. Based on these data, the dietary needs for 1 or more of the 5 B vitamins are greater than current NRC (1998) estimates, particularly in pigs expressing a high rate of lean tissue growth. The greater need for these vitamins is not associated with greater dietary energy intake or body energy accretion rate but is potentially due to shifts in the predominant metabolic pathways.