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Amino acid requirements for maturation of rhesus monkey (Macacca mulatta) oocytes in culture.

This study evaluated the effects of different amino acid formulations on supporting meiotic and cytoplasmic maturation of rhesus monkey (Macacca mulatta) oocytes in vitro. Five hundred and forty-six cumulus-oocyte complexes (COCs) aspirated from unstimulated adult monkey follicles (> or = 1000 microm in diameter) were cultured in either modified Connaught Medical Research Laboratories 1066 medium (mCMRL-1066) or in one of eight chemically defined media (modified basic medium 5 supplemented with 5.5 mmol glucose l(-1), 0.003 mmol pantothenic acid l(-1) and different amino acid formulations) as below: (1) modified basic medium 5 (mBM5) containing no amino acid; (2) mBM5 + 0.2 mmol glutamine l(-1); (3) mBM5 + 11 amino acids from hamster embryo culture medium 6 (HECM-6) (11 AA); (4) mBM5 + Eagle's non-essential amino acids (NEA); (5) mBM5 + NEA + 0.2 mmol glutamine l(-1); (6) mBM5 + Eagle's essential amino acids (EA) without glutamine; (7) mBM5 + EA + 0.2 mmol glutamine l(-1); (8) mBM5 + Eagle's 20 amino acids (20 AA) + 0.2 mmol glutamine l(-1); and (9) mCMRL-1066 (control). All media contained FSH, LH, oestradiol and progesterone. After maturation, mature oocytes were subjected to the same fertilization and embryo culture procedures. COCs matured in treatment 5 had greater potential to progress to metaphase II (66%; P < 0.05) than did those in treatments 1 (37.3%), 2 (48.3%), 3 (41%), 6 (41%) and 9 (43%). Oocytes matured in treatment 8 had the best morula (53%) and blastocyst (18%) developmental responses (P < 0.05). The lowest (P < 0.05) morula and blastocyst developmental responses were obtained from COCs matured in treatments 1 (0%) and 6 (8%). The other media supported intermediate embryonic development (range 11-38% of morula and blastocyst). These results indicate that the choice of amino acids affects the competence of oocyte maturation and that Eagle's 20 AA with 0.2 mmol glutamine l(-1) is more efficient than the other amino acid formulations for maturation of rhesus monkey oocytes.

Amino Acids↗

Evaluation of prenatal vitamin-mineral supplements.

Nutritional requirements of pregnancy are reviewed, and guidelines for evaluating prenatal vitamin-mineral supplements are provided. Daily antepartum supplementation of 0.4-0.8 mg of folic acid and 30-60 mg of elemental iron is currently recommended, although the lower ends of these ranges may be most appropriate. Dietary intake of these nutrients is likely to be inadequate without supplementation, and their importance is well established. Requirements for other minerals and vitamins are not well established, and there is no consensus on the need for supplementation. However, available data suggest that prenatal supplements should probably contain other nutrients; pyridoxine hydrochloride, cholecalciferol, vitamin E, pantothenic acid, calcium, magnesium, zinc, copper, and possibly selenium should be considered. Interactions among the minerals and vitamins commonly found in prenatal supplements may affect the absorption of various nutrient components. Thus, very high or low levels of certain nutrients should be avoided. The chemical form of minerals should also be considered. Products should have demonstrated bioavailability for iron, zinc, and other components that are subject to bioavailability problems. Use of low-potency product that contains a wide range of vitamins and minerals appears to be the most prudent approach to prenatal vitamin and mineral supplementation.

Absorption↗

Post-translational disulfide modifications in cell signaling--role of inter-protein, intra-protein, S-glutathionyl, and S-cysteaminyl disulfide modifications in signal transmission.

Cell signaling entails a host of post-translational modifications of effector-proteins. These modifications control signal transmission by regulating the activity, localization or half-life of the effector-protein. Prominent oxidative modifications induced by cell-signaling reactive oxygen species (ROS) are cysteinyl modifications such as S-nitrosylation, sulfenic acid and disulfide formation. Disulfides protect protein sulfhydryls against oxidative destruction and simultaneously influence cell signaling by engaging redox-regulatory sulfhydryls in effector-proteins. The types of disulfides implicated in signaling span (1) protein S-glutathionylation, e.g. as a novel mode of Ras activation through S-glutathionylation at Cys-118 in response to a hydrogen-peroxide burst, (2) intra-protein disulfides, e.g. in the regulation of the stability of the protein phosphatase Cdc25C by hydrogen-peroxide, (3) inter-protein disulfides, e.g. in the hydrogen peroxide-mediated inactivation of receptor protein-tyrosine phosphatase alpha (RPTPalpha) by dimerization and (4) protein S-cysteaminylation by cystamine. Cystamine is a byproduct of pantetheinase-catalyzed pantothenic acid recycling from pantetheine for biosynthesis of Coenzyme A (CoA), a ubiquitous and metabolically indispensable cofactor. Cystamine inactivates protein kinase C-epsilon (PKCepsilon), gamma-glutamylcysteine synthetase and tissue transglutaminase by S-cysteaminylation-triggered mechanisms. The importance of protein S-cysteaminylation in signal transmission in vivo is evident from the ability of cystamine administration to rescue the intestinal inflammatory-response deficit of pantetheinase knockout mice. These mice lack the predominant epithelial pantetheinase isoform and have sharply reduced levels of cystamine/cysteamine in epithelial tissues. In addition, intraperitoneal administration of cystamine significantly delays neurodegenerative pathogenesis in a Huntington's disease mouse model. Thus, cystamine may serve as a prototype for the development of novel therapeutics that target effector-proteins regulated by S-cysteaminylation.

Animals↗

Tests of the adaptive modulation hypothesis for dietary control of intestinal nutrient transport.

According to the adaptive modulation hypothesis, an intestinal transporter should be repressed when its biosynthetic and other costs (of maintenance) exceed the benefits it provides. This leads to two contrasting predictions: transport of a sugar or amino acid worth calories should tend to be increased by its substrate, and transport of a vitamin should be modulated downwards by its substrate and upmodulated in deficiency. In a test of the first prediction, omnivorous desert iguanas eating alfalfa pellets (a high-carbohydrate diet) were compared with desert iguanas eating mealworms (a low-carbohydrate, higher-protein diet). In accord with the prediction, intact intestinal sleeves from the former group had higher rates of carrier-mediated D-glucose uptake/centimeter across the brush border than sleeves from the latter group. But in contrast to the first prediction, mealworm eaters had lower (not higher) proline uptake rates, and the ratio of glucose/proline uptake in the two groups did not differ. I review similar tests in 12 other species and show that overall the hypothesis has been quite robust with regard to the first prediction. Cases in which the hypothesis is rejected may reflect complications associated with changes in other dietary factors or phylogenetic constraints. In a test of the second prediction, uptake of the water-soluble vitamin choline was not increased in choline-deficient chicks, nor was it decreased in adults that have no dietary requirement for choline. I review similar tests for four other vitamins and five essential minerals. Dietary control of transport of the minerals and two of the vitamins seems to be in accord with the hypothesis. But transport rate for three vitamins (choline, pantothenic acid, ascorbic acid) seems not to be increased in deficiency. The best explanation seems to be that vitamin transport is modulated only if it is primarily by a carrier-mediated pathway.

Adaptation, Physiological↗

Vitamins and cancer prevention: issues and dilemmas.

Vitamins are a class of organic compounds that are components of an adequate diet. They or their derivatives function as coenzymes, cellular antioxidants, and/or regulators of gene expression. Fourteen vitamins are recognized in human nutrition (Vitamins A, D, E, K, B1, B2, B6, B12, C, niacin, folacin, pantothenic acid, biotin, choline), with deficiencies or excesses in intake leading to changes in protein, nucleic acid, carbohydrates, fat and/or mineral metabolism. Thus, the integrity of physiological systems, including those associated with detoxification, cellular repair, immune processes, and neural and endocrine function, depends upon the nutritional and vitamin status of the host. For these reasons, it may be anticipated that the adequacy of the vitamin supply to cells and tissues would affect the development, progress, and outcome of cancers. In this review, the definition and functions of and requirements and recommended allowance for vitamins are discussed briefly before exploring the evidence, largely from studies in experimental animals, that indicates the nature of the link between vitamins and cancer. Although evidence based on studies in animal systems reveals that vitamin intake and status can modulate the outcome of experimental carcinogenesis, the findings are often conflicting and difficult to interpret. Furthermore, it is not yet possible to develop a suitable prediction of the role of the individual vitamins in tumor development. The significance of these observations for human nutrition and cancer prevention, particularly in reference to ascorbic acid (vitamin C), vitamin E, and B-complex vitamins is considered. Vitamin A and retinoid compounds are discussed elsewhere in the symposium. The many popular misconceptions and unsound advice concerning vitamins and health, including "fake" vitamins-pangamic acid ("vitamin B15") and laetrile ("vitamin B17")-are also discussed. On the basis of current evidence, it would be inappropriate to recommend either substantial changes in habitual vitamin intakes, as provided by an adequate, well-balanced diet, or promotion of megavitamin intakes, as a means of reducing risk from cancers in the human population. However, a prudent approach toward diet and food habits, as a means of better optimizing the health consequences of our complex lifestyle is to be recommended.

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Determination of water-soluble vitamins in soft drinks and vitamin supplements using capillary electrophoresis.

A method for the determination of six water-soluble vitamins based on capillary electrophoresis (CE) operated in micellar mode was developed. Thiamine hydrochloride (vitamin B1), riboflavin (vitamin B2), pyridoxine hydrochloride (vitamin B6), pantothenic acid (vitamin B5), nicotinamide (vitamin B3), and cobalamin (Vitamin B12) could be separated in a single run. All CE parameters such as buffer composition and operation temperature were optimized in order to achieve better separation. Relative standard deviations (RSDs) of the described method ranged from 1.08 to 3.68% (intra-day precision) and 1.26 to 3.35% (inter-day precision). The method was then used for measuring various soft drinks and vitamin supplements directly without any step of sample cleanup. The determination of niacin was successful for all samples tested, reaching recoveries near 100%. Riboflavin and pyridoxine were quantified successfully in some but not all samples. Therefore, an evaluation on a case-by-case basis is mandatory. When applicable, this method provides a fast, accurate, simple, and inexpensive way to quantify selected vitamins, and is therefore well suited for routine analysis in soft drink industry.

Beverages↗

Production of vitamins, coenzymes and related biochemicals by biotechnological processes.

Vitamins and related biofactors belong to those few chemicals with a direct positive appeal to people. There is indeed a large need for extra vitamins, other than those derived from plant and animal food sources, due to unbalanced food habits or processing, food shortage or disease. Added vitamins are now either prepared chemically or biotechnologically via fermentation or bioconversion processes. Several vitamins and related biofactors are now only or mainly produced chemically (vitamin A, cholecalciferol (D3), tocopherol (E), vitamin K2, thiamine (B1), niacin (PP or B3), pantothenic acid (B5), pyridoxine (B6), biotin (H or B8), folic acid (B9) or via extraction processes (beta-carotene or provitamin A, provitamin D3, tocopherol, vitamin F-group). However, for several of these compounds microbiological or algal methods also exist or are rapidly emerging. Others are produced practically exclusively via fermentation (ergosterol or provitamin D2, riboflavin (B2), cyanocobalamin (B12), orotic acid (B13), vitamin F-group, ATP, nucleosides, coenzymes, etc. or via microalgal culture (beta-carotene, E, F). Both chemical and microbial processes are run industrially for vitamin B2 while vitamin C (ascorbic acid) is produced via a combination of chemical reactions and fermentation processes. A survey is given here of the current state of vitamin production, with emphasis on developments and strategies for improved biotechnological production and its significance, as compared to existing chemical processes. The screening or construction of vitamin hyperproducing microbial strains is a difficult task; pathway elucidation and metabolic (de)regulation need further study; r-DNA technology has only recently been introduced; improved fermentation processes and immobilised biocatalysts bioconversions for the synthesis of chiral vitamin compounds or intermediates or derivatives are gaining importance; the recovery and purification of these vitamin compounds from their fermentation broths remains equally complex.

Biotechnology↗

Mutations in ADE3 reduce the efficiency of the omnipotent suppressor sup45-2.

Mutations in a known yeast gene, ADE3, were shown to act as an antisuppressor, reducing the efficiency of the omnipotent suppressor, sup45-2. The ADE3 locus encodes the trifunctional enzyme C1-tetrahydrofolate synthase, which is required for the biosynthesis of purines, thymidylate, methionine, histidine, pantothenic acid and formylmethionyl-tRNA(fMet. The role of this enzyme in translational fidelity had not previously been suspected.

Aminohydrolases↗

Chemically defined media for growing anaerobic bacteria of the genus Veillonella.

A chemically defined medium for Veillonella parvula and V. alcalescens is described. Some nutritional aspects of the two strains used were examined: the optimum concentration of reducing agents, the requirements for amino acids, diamines, vitamins and other growth factors, and the conditions needed for well balanced nutrition. No specific requirements for single amino acids were observed. A combination of L-cysteine, DL-aspartic acid, L-glutamic acid, L-serine and L-tyrosine, promoted growth. In V. alcalescens, serine could substitute both arginine and tryptophan (or histidine). No growth was obtained with ammonium salts as the sole N source. Decarboxylation of L-ornithine, L-lysine and L-arginine was not demonstrated in the Veillonella parvula strain, which required putrescine or cadaverine for growth. Spermine, spermidine, L-lysine, L-ornithine and L-arginine, could not substitute putrescine in Veillonella parvula. Veillonella alcalescens, which does not require putrescine in the medium, was able to decarboxylate L-ornithine while forming putrescine.

4-Aminobenzoic Acid↗

The use of Weibull distribution to describe the in vivo absorption kinetics.

The drug in vivo absorption rate reflects the distribution of drug molecules absorption times. For the latter the Weibull distribution is suggested and some examples taken from the literature are used to support the proposal. Cumulative absorption data for theophylline in man and pantothenic acid in rats are linearized in the double log coordinates, and the Weibull absorption model parameters are estimated by linear regression. The mean absorption time values calculated from these parameters are in close agreement with those assessed by the model-independent method.

Absorption↗

[Survey of the present-day supply of fortified food products in Germany].

A survey and critical evaluation of the present-day supply of fortified common food products on the German market is presented concerning products, nutrients and amounts for fortification. The data were collected from the original food labels by personal informations from the manufacturers (40 asked, 68% answered) and by a local market survey in Dortmund (spring 1994). A total of 288 fortified food products (78 manufacturers) were found out of 6 different food products (78 manufacturers) were found out of 6 different food categories (manufacturers/products): beverages (26/95), sweets (24/57), cereals (5/53), milk products (7/35), powdered instant beverages (10/31), ready-to-eat meals (6/17). Sugar was added to 56% of the fortified products. A total of 10 vitamins (E, B1, B2, B6, B12, C, niacin, folate, biotin, pantothenic acid) and 7 minerals (Na, K, Cl, Ca, P, Mg, Fe) were used for fortification. The number of nutrients used for fortification in single products ranged from 1 (94 products) to 16 (3 products). The amounts used for fortification per average portion varied considerably among the different products and the different nutrients in the food categories. In a considerable number of cases, amounts for fortification of more than 100% (maximum 660%) of the recommended daily dose of a nutrient (EC-directive for nutrition labelling) have been observed. Relatively to the contribution of a portion to meet the energy requirement, the fortification of cereals and milk-products (about 30%) as well as of sweets (about 10%) could be rated as acceptable whereas the fortification of beverages (about 50-100%) was overdone. The present-day supply of fortified food is extremely heterogenous from the qualitative and quantitative point of view. Therefore, it is rather difficult for the consumer to reach an overall nutrient intake that is in accordance with the recommendations. The specific requirements of children, who are a preferred group for food advertising, are not at all considered adequately.

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