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Preventive effects of L-ascorbic acid and calcium pantothenate against photosensitive actions induced by pheophorbide alpha and hematoporphyrin.

The preventive effects of L-ascorbic acid (V.C) and pantothenic acid (PaA) against photosensitized hemolysis in the presence of pheophorbide a or hematoporphyrin were investigated in vitro. The development of hematoporphyrin-induced photosensitivity and the preventive effects of V.C and PaA were also investigated in male mice. V.C and PaA exerted preventive effects against photosensitized hemolysis in the presence of pheophorbide a or hematoporphyrin. In such cases, V.C and PaA had not only an additive effect but also a synergistic one. The time required for the photosensitized acute shock-like death (i.e., survival time), was significantly extended by simultaneous administration of V.C and PaA. This supports the findings of in vitro experiments.

Animals↗

Cofactor engineering of intracellular CoA/acetyl-CoA and its effect on metabolic flux redistribution in Escherichia coli.

Coenzyme A (CoA) and its thioester derivatives are important cofactors participating in over 100 different reactions in intermediary metabolism of microorganisms. The time profiles of intracellular CoA and acetyl-CoA levels were studied in an aerobic batch reactor. The CoA level starts at a high value and falls off gradually over the exponential and stationary growth phases, reaching negligible levels at the end of 24h. The acetyl-CoA level, on the other hand, increases initially reaching a maximum and decreases gradually reaching negligible levels after 24h. Overexpressing one of the upstream rate-controlling enzyme the pantothenate kinase with simultaneous supplementation of the precursor pantothenic acid to the culture medium increased the intracellular CoA/acetyl-CoA levels. It was found that supplementation of the precursor pantothenic acid is essential to increase CoA/acetyl-CoA levels. A 10-fold increase in CoA level was observed upon this overexpression in complex medium. Acetyl-CoA levels also increased (5-fold) but not as much as CoA, leaving much of the CoA in free unacetylated form. The increase in intracellular CoA/acetyl-CoA levels led to an increase in carbon flux to the acetate production pathway leading to formation of more acetate in complex medium, whereas no such change in metabolite redistribution was observed in minimal medium.

Acetates↗

Water-soluble vitamins.

Simultaneous Determination of Vitamins.--Klejdus et al. described a simultaneous determination of 10 water- and 10 fat-soluble vitamins in pharmaceutical preparations by liquid chromatography-diode-array detection (LC-DAD). A combined isocratic and linear gradient allowed separation of vitamins in 3 distinct groups: polar, low-polar, and nonpolar. The method was applied to pharmaceutical preparations, fortified powdered drinks, and food samples, for which results were in good agreement with values claimed. Heudi et al. described a separation of 9 water-soluble vitamins by LC-UV. The method was applied for the quantification of vitamins in polyvitaminated premixes used for the fortification of infant nutrition products. The repeatability of the method was evaluated at different concentration levels and coefficients of variation were <6.5%. The concentrations of vitamins found in premixes with the method were comparable to the values declared. A disadvantage of the methods mentioned above is that sample composition has to be known in advance. According to European legislation, for example, foods might be fortified with riboflavin phosphate or thiamin phosphate, vitamers which are not included in the simultaneous separations described. Vitamin B2.--Viñas et al. elaborated an LC analysis of riboflavin vitamers in foods. Vitamin B2 can be found in nature as the free riboflavin, but in most biological materials it occurs predominantly in the form of 2 coenzymes, flavin mononucleotide (FMN) and flavin-adenine dinucleotide (FAD). Several methods usually involve the conversion of these coenzymes into free riboflavin before quantification of total riboflavin. According to the authors, there is growing interest to know flavin composition of foods. The described method separates the individual vitamers isocratically. Accuracy of the method is tested with 2 certified reference materials (CRMs). Vitamin B5.-Methods for the determination of vitamin B5 in foods are limited because of their low sensitivity and poor selectivity. Pakin et al. proposed a post-column derivatization of pantothenic acid as a fluorescent compound and used this principle in a specific and sensitive method for the determination of free and bound pantothenic acid in a large variety of foods. A French laboratory invited European laboratories to participate in a series of collaborative studies for this method, which will be carried out in 2005/2006. A more sophisticated method was described by Mittermayer et al. They developed an LC-mass spectrometry (LC/MS) method for the determination of vitamin B5 in a wide range of fortified food products. Application of the method to various samples showed consistent results with those obtained by microbiology. Vitamin B6.-Method 2004.07, an LC method for the analysis of vitamin B6 in reconstituted infant formula, was published by Mann et al. In contrast with this method, which quantifies vitamin B6 after converting the phosphorylated and free vitamers into pyridoxine, Viñas et al. published an LC method which determines 6 vitamin B6 related compounds, the 3 B6 vitamers, their corresponding phosphorylated esters, and a metabolite. Accuracy was determined using 2 CRMs. Results were within the certified ranges. Vitamin C.-Franke et al. described an extensive study to vitamin C and flavonoid levels of fruits and vegetables consumed in Hawaii. Vitamin C was determined by measuring ascorbic acid in its reduced state by LC and coulometric detection along with UV absorbance detection at 245 nm. No attempts were made to assess levels of dehydroascorbic acid. Most recent research revealed that cell uptake of dehydroascorbic acid is unlikely to play a major role, which may explain the very low vitamin C activity of orally administered L-dehydroascorbic acid in rats. The food levels found by Franke et al. are variably lower, higher, or equal in comparison to other studies. Iwase described a method for the determination of ascorbic acid in foods using L-methionine for the pre-analysis sample stabilization. Electrochemical detection was used for the quantification. Traditionally, metaphosphoric acid was proven to be a useful dissolving agent for the determination of ascorbic acid. However, it dissolves in water very slowly, it is hygroscopic, and accurate weighing is not easy. Adjustment at pH 2-3 takes a long time. It appeared to be possible to replace metaphosphoric acid by 0.2% phosphoric acid. Methionine played an important role on the stability of ascorbic acid. The method seemed to be applicable to the routine analysis of ascorbic acid in foods. Folic Acid.-Microbiological analysis of total folate in foods is often considered as the golden standard compared to other methods based on, for example, LC. Koontz et al. showed results of total folate concentrations measured by microbiological assay in a variety of foods. Samples were submitted in a routine manner to experienced laboratories that regularly perform folate analysis fee-for-service basis in the United States. Each laboratory reported the use of a microbiological method similar to the AOAC Official Method for the determination of folic acid. Striking was, the use of 3 different pH extraction conditions by 4 laboratories. Only one laboratory reported using a tri-enzyme extraction. Results were evaluated. Results for folic acid fortified foods had considerably lower between-laboratory variation, 9-11%, versus >45% for other foods. Mean total folate ranged from 14 to 279 microg/100 g for a mixed vegetable reference material, from 5 to 70 microg/100 g for strawberries, and from 28 to 81 microg/100 g for wholemeal flour. One should realize a large variation in results, which might be caused by slight modifications in the microbiological analysis of total folate in foods or the analysis in various (unfortified) food matrixes. Furthermore, optimal combination of enzymes and reaction conditions may vary depending on the composition of the food. Padrangi and Laborde showed recently that treatment with alpha-amylase had no significant effect on measured folate in spinach, although addition of protease significantly increased the release of folate. LC/MS applications gain increasing attention because of their specificity. Rychlik used stable isotope dilution assays for the determination of the folate content of broccoli and bread. Compared to data in the literature and food data bases, amounts were significantly lower. Pawlosky et al., however, found comparable values for 5-methyltetrahydrofolic acid and folic acid by HPLC analysis with fluorescent detection and HPLC/MS. Among samples analyzed were CRMs and broccoli. Besides folic acid, other water-soluble vitamins were also determined by LC/MS/MS by Leporati et al. The method was applied to the quantitative analysis of the natural content of vitamins in typical Italian pasta samples, as well as in fortified pasta samples produced for the U.S. market. Biotin.-A paper from Staggs et al. included the assertion that existing biotin data in food composition tables are inaccurate because the majority are based on bioassays with all relevant disadvantages. Data in most cases are overestimated with consequences for recommendations for dietary biotin intake. An HPLC/avidin-binding assay was used to analyze 87 foods to support the hypothesis mentioned.

Chemistry Techniques, Analytical↗

Lactation performance by dairy cows fed supplemental biotin and a B-vitamin blend.

The objective of Trial 1 was to evaluate in dairy cows the effects of dietary supplementation with biotin and a B-vitamin blend on dry matter intake (DMI), milk yield, composition and component yields, total tract nutrient digestion, and plasma metabolites. Twenty-four multiparous Holstein cows averaging 46 +/- 8 d in milk at trial initiation were randomly assigned to treatments in a replicated 4 x 4 Latin square design with 28 d periods. The four treatments were: 1) a control diet (C) with no supplemental B-vitamins; 2) C plus supplemental biotin at 20 mg/d (B); 3) C plus supplemental thiamin (150 mg/d), riboflavin (150 mg/d), pyridoxine (120 mg/d), B12 (0.5 mg/d), niacin (3000 mg/d), pantothenic acid (475 mg/d), folic acid (100 mg/d), and biotin (20 mg/d) (BBVIT1X); 4) C plus supplemental thiamin (300 mg/d), riboflavin (300 mg/d), pyridoxine (240 mg/d), B12 (1.0 mg/d), niacin (6000 mg/d), pantothenic acid (950 mg/d), folic acid (200 mg/d), and biotin (40 mg/d) (BBVIT2X). Intake of DM was increased 0.7 kg/d for B vs. C and BBVIT1X and 1.3 kg/d for B vs. BBVIT2X. Milk yield was increased 1.7 kg/d for B vs. C. For BBVIT1X, milk yield was similar to B and BBVIT2X and tended to be higher than C. Yields of milk protein and lactose but not fat were higher for B than C. For BBVIT1X, milk component yields were similar to B and tended to be higher than C, with the exception of lactose yield where BBVIT1X was higher than C. The objective of Trial 2 was to evaluate DMI and milk yield, composition and component yields by dairy cows fed diets supplemented with either 40 mg/d biotin or the B-vitamin blend (BBVIT1X) compared to cows supplemented with 20 mg/d dietary biotin. Neither the 40 mg/d biotin treatment nor the B-vitamin blend enhanced lactation performance over the 20 mg/d biotin treatment. Biotin efficacy in short-term trials suggests that biotin may improve milk yield directly via effects on intake and (or) nutrient metabolism rather than indirectly via improved hoof health. More research is needed to determine the mode of action for supplemental dietary biotin.

3-Hydroxybutyric Acid↗

The effect of Schistosoma mansoni infection, and of oxamniquine, a new antischistosomal drug on some vitamins in the livers of hamsters.

The hepatic levels of thiamine, pantothenic acid and niacin in hamsters infected with S. mansoni were depressed, while the levels of riboflavin and pyridoxine remained unchanged. A single dose of oxamniquine, 10-50 mg/kg, brought the levels of thiamine, pantothenic acid and niacin to values not significantly different from those in non-infected controls, but lowered the concentrations of riboflavin and pyridoxine in infected hamsters.

Animals↗

Crystal structures of Fms1 and its complex with spermine reveal substrate specificity.

Fms1 is a rate-limiting enzyme for the biosynthesis of pantothenic acid in yeast. Fms1 has polyamine oxidase (PAO) activity, which converts spermine into spermidine and 3-aminopropanal. The 3-aminopropanal is further oxidized to produce beta-alanine, which is necessary for the biosynthesis of pantothenic acid. The crystal structures of Fms1 and its complex with the substrate spermine have been determined using the single-wavelength anomalous diffraction (SAD) phasing method. Fms1 consists of an FAD-binding domain, with Rossmann fold topology, and a substrate-binding domain. The active site is a tunnel located at the interface of the two domains. The substrate spermine binds to the active site mainly via hydrogen bonds and hydrophobic interactions. In the complex, C11 but not C9 of spermine is close enough to the catalytic site (N5 of FAD) to be oxidized. Therefore, the products are spermidine and 3-aminopropanal, rather than 3-(aminopropyl) 4-aminobutyraldehyde and 1,3-diaminoprone.

Amino Acid Sequence↗

Metabolism of pantethine in cystinosis.

D-Pantethine is a conjugate of the vitamin pantothenic acid and the low-molecular-weight aminothiol cysteamine. Pantethine is an experimental hypolipemic agent and has been suggested as a source of cysteamine in the treatment of nephropathic cystinosis. We treated four cystinotic children with 70-1,000 mg/kg per d oral D-pantethine and studied its metabolism. Pantethine was rapidly hydrolyzed to pantothenic acid and cysteamine; we could not detect pantethine in plasma after oral administration. The responsible enzyme, "pantetheinase," was highly active in homogenates of small intestinal mucosa and plasma. The Michaelis constant of the rat intestinal enzyme was 4.6 microM and its pH profile showed a broad plateau between 4 and 9. Pantothenate pharmacokinetics after orally administered pantethine followed an open two-compartment model with slow vitamin elimination (t1/2 = 28 h). Peak plasma pantothenate occurred at 2.5 h and levels over 250 microM were seen at 300 times normal. Apparent total body storage of pantothenate was significant (25 mg/kg), and plasma levels were elevated threefold for months after pantethine therapy. Plasma cysteamine concentrations after pantethine were similar to those reported after equivalent doses of cysteamine. However, at best only 80% white blood cell cystine depletion occurred. We conclude that pantethine is probably less effective than cysteamine in the treatment of nephropathic cystinosis and should only be considered in cases of cysteamine intolerance. Serum cholesterol was decreased an average of 14%, which supports the potential clinical significance of pantethine as a hypolipemic agent. Rapid in vivo hydrolysis of pantethine suggests that pantothenate or cysteamine may be the effectors of its hypolipemic action.

Adolescent↗

Cellular uptake of biotin: mechanisms and regulation.

This review describes our knowledge of biotin transport in the small intestine of humans and other mammals and presents recent findings in the area. Previous studies have shown that biotin transport across the brush border membrane of the small intestinal absorptive cells occurs via a carrier-mediated, Na+ gradient-dependent, electroneutral mechanism. Exit of biotin out of the enterocyte, i.e., transport across the basolateral membrane, also occurs via a carrier-mediated process, but the process is Na+ independent and electrogenic. Recent studies from our laboratory have shown that the uptake process of biotin in Caco-2 cells, a human-derived cultured intestinal epithelial cell line, are under the cellular regulation of both a protein kinase C- and a Ca/calmodulin-mediated pathway. In addition, the uptake process is shared by another water-soluble vitamin, pantothenic acid. For the first time, other recent studies have detected the existence of a Na+-dependent, carrier-mediated mechanism for biotin uptake at the apical membrane of colonocytes, which could theoretically mediate absorption of the biotin synthesized by colonic microflora. This system was again found to be shared by pantothenic acid, which is also synthesized by the normal microflora of the large intestine.

Animals↗

[Studies on the vitamin requirement and vitamin supply in fattening turkeys. 2. Checking the vitamin supplements to state-approved mixed animal feed for fattening turkeys].

In turkey fattening experiments, the vitamin A, vitamin D3, vitamin B6, vitamin B12, niacine, pantothenic acid, folic acid, biotine and choline chloride additions to mixed feeds for fattening turkeys were checked. The vitamin A requirement of the fattening turkey is met, in all stages of growth, by adding 3,000 I.U./kg feed. For vitamin D3 the necessary supplements are 1,000 and 500 I.U./kg for starter feed and fattening mixes, respectively. The requirements for vitamins B2, B6, B12, niacine, pantothenic acid, folic acid and biotine of the fattening turkey are covered by the respective amounts contained in the ration components. To ensure adequate choline supply (optimum gain, small losses) during the starter period, the addition of 500 mg/kg feed proved necessary. On the basis of the results presented, recommendations are given for requirement norms and mixed feed supplementation that have been fully implemented in the meantime.

Animal Feed↗

Measurement of the bioavailability of iron, using the rat hemoglobin repletion test.

Variations in the AOAC official first action rat hemoglobin repletion test for iron were studied. These changes included (1) use of a simplified basal diet to eliminate ingredients which sometimes contribute too much iron; (2) increased fortification of the basal diet with vitamin E, pantothenic acid, and pyridoxine; (3) increased dietary copper; (4) variations in the carbohydrate source in the basal diet; (5) changes in the length of the depletion and repletion periods; and (6) comparison of prophylactic and curative procedures. The changes yielded results comparable to those obtained with the present official method. Further study may reveal that the depletion period can be shortened or eliminated. To fully meet the rat's vitamin requirements, increased levels of vitamin E, pantothenic acid, and pyridoxine are recommended. It is further recommended that the present method remain in official first action status, and that study be continued.

Animals↗

Composition of vitamin supplements in Spanish poultry diets.

1. The composition of 106 vitamin supplements used in about 85% of the Spanish poultry production diets were studied. Vitamin supplements were grouped by production classes and, for broilers and pullets, also by feeding periods. 2. Four vitamins (niacin, alpha-tocopherol, pantothenic acid, and riboflavin) comprised over 87% of the vitamin supplements by weight (choline excluded), whereas alpha-tocopherol and retinol represented from 51% to 60% of the total vitamin cost. 3. The highest and lowest vitamin supplementation rates were for broilers in the starter and withdrawal periods (106 and 44 mg/kg, respectively) and the mean values for breeders, pullets and layers were 104, 58, and 48 mg/kg, respectively. 4. Supplements with higher vitamin contents showed less variability in their composition. Retinol, cholecalciferol, riboflavin and pantothenic acid showed the lowest variability within supplements (6% to 36% CV), whereas alpha-tocopherol, menadione, thiamin and biotin showed the highest (40% to 224% CV). 5. Vitamin supplementation rates were compared with requirements, taking into account the dietary contribution. In general, vitamin fortification exceeded the NRC recommendations, using a high safety margin for some vitamins such as vitamin A (from 2.6 to 7.8) and for some poultry classes such as breeders (3.2).

Animal Feed↗

Vitamin intake in Japanese women college students.

The Standard Food Tables of Japanese Foods was newly revised in 2000, and contains information on all of the vitamins except biotin. Thus, we carried out a survey of vitamin intake in Japanese women who were university seniors majoring a dietitian course. The subjects (n = 33) consumed self-selected foods, and food intake was recorded by the weight method. We calculated the vitamin intake except for biotin from the food records using the Standard Food Tables of Japanese Foods. In terms of daily intake, vitamin A was 705+/-435 microg (mean+/-SD), vitamin D 6+/-8 microg, vitamin E 7.7+/-3.0 mg, vitamin K 191+/-156 microg, vitamin B1 0.7+/-0.3 mg (0.43+/-0.15 mg/1,000 kcal), vitamin B2 1.1+/-0.4 mg (0.65+/-0.18 mg/1,000 kcal), vitamin B6 0.9+/-0.4 mg (0.017+/-0.005 mg/g protein), vitamin B12 4.4+/-4.1 microg, niacin equivalent 23+/-7 mg (14.4+/-4.9 mg/1,000 kcal), pantothenic acid 4.6+/-1.4 mg, folic acid 267+/-115 microg, and vitamin C 73+/-38 mg. All of these averages were around the Japanese Recommended Dietary Allowance (RDA) for level "III (preferable)" of physical activity. Major vitamin A resources were vegetables; vitamin D resources, fish; vitamin E resources, fats and oils and vegetables; vitamin K resources, vegetables; vitamin B1 resources, cereals and animal meats; vitamin B2 resources, various foods; vitamin B6 resources, cereals, vegetables, fish, and animal meats; vitamin B12 resources, fish; niacin equivalent resources, fish, animal meats, and cereals; pantothenic acid resources, various foods; folic acid resources, vegetables; and vitamin C resources, vegetables and potatoes. From this survey, it was found that Japanese women college students consumed many kinds of food, and therefore, their vitamin nutrition was good as compared to the RDA values for level III of physical activity; however, their energy intake (1,622+/-377 kcal) was lower than the RDA for level III (2,050 kcal/d). Their strength of physical activity would be level I. Therefore, in consideration of their lifestyle, their energy intakes is considered adequate. In conclusion, a problem for student lifestyle is a shortage of food intake due to lack of exercise.

Adult↗

Influence of pre-, pro-, and synbiotics on the intestinal availability of different B-vitamins.

This study was conducted with 16 male growing pigs (eight pigs with an end-to-end ileo-rectal anastomosis, eight intact pigs) to investigate in metabolic trials the influence of different pre-, pro-, and synbiotics on the precaecal and faecal digestibility of thiamin, riboflavin, pantothenic acid and biotin. A basic experimental diet was supplemented with either pre-, pro-, or synbiotic additives as follows: lactulose (2%), inulin (2%), mannanoligosaccharides (0.3%), Enterococcus faecium DSM 10663 (8 x 10(9) cfu/kg), Enterococcusfaecium DSM 7134 (5 x 10(8) cfu/kg), inulin + Enterococcusfaecium DSM 10663, mannanoligosaccharides + Enterococcus faecium DSM 7134. The precaecal and faecal digestibilities of the investigated B-vitamins were not affected significantly by the administered pre-, pro-, and synbiotics independent of the application period. A higher faecal vitamin excretion vs. the precaecal indicated that thiamin and biotin were synthesized in the colon. On the contrary, for riboflavin and pantothenic acid a higher precaecal vitamin flow compared to the faecal excretion was detected indicating to a certain extent an absorption in the colon.

Anastomosis, Surgical↗

Vitamin and proximate composition of fast-food fried chicken.

As part of the USDA's effort to obtain accurate data for nutrient composition handbooks, we determined proximate composition and vitamin (thiamin, riboflavin, niacin, folacin, pantothenic acid, and vitamins B-6 and B-12) content of fast-food fried chicken. Ninety-four samples were purchased from six chains in 10 cities throughout the United States. Vitamin content of breast and thigh meat portions differed, and slight differences in composition among brands were noted. Thiamin values for breast and thigh muscle, respectively, ranged from 0.062 to 0.136 and from 0.078 to 0.122 mg/100 gm; riboflavin, from 0.176 to 0.205 and 0.249 to 0.328 mg/100 gm; niacin, from 6.32 to 8.46 and 3.86 to 5.57 mg/100 gm; vitamin B-6, from 0.315 to 0.390 and 0.189 to 0.271 mg/100 gm; folacin, from 4.2 to 6.9 and 5.00 to 8.00 micrograms/100 gm; vitamin B-12, from 0.331 to 0.459 and 0.382 to 0.596 micrograms/100 gm; and pantothenic acid, from 1.48 to 1.68 and 1.58 to 1.77 mg/100 gm. Pressure-fried samples tended to have higher percentage moisture and lower percentage fat than crispy, open-vat fried samples. Fat ranged from about 15% to 20%.

Animals↗

Effect of alpha-tocopherol, pyridoxine and dexpanthenol on the stress in crease of nonesterified fatty acids levels in the brain.

The supposed antistress effect of vitamins-alpha-tocopherol, pyridoxine and dexpanthenol (pantothenic acid precursor)--was followed on the model of nociceptive stress in laboratory rats. The decrease of the stress enhancement of nonesterified fatty acids (NEFA), estimated in the brain cortex, hypothalamus and the brain stem, was taken for the indicator of the antistress effect. Nonesterified fatty acids were determined with the help of gas chromatography following the separation performed by thin layer chromatographic method. Five-day application of alpha-tocopherol acetate (per os, 300 mg.kg-1) led to a decrease of the stress enhancement of arachidonic acid level in the brain stem.

Animals↗