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Involvement of a recombination repair function in disciplined cell division of Micrococcus radiodurans.

When a culture of the temperature-sensitive DNA mutant Micrococcus radiodurans tsI is irradiated with a sublethal dose of ultraviolet or ionizing radiation and is plated immediately, all the bacteria give rise, after 36 h incubation, to colonies identical to those derived from unirradiated bacteria. However, when the irradiated population is held at its restrictive temperature (39 degrees C) (restrictive temperature holding) for 3 h before being plated, less than 0-1% of the surviving bacteria give rise to normal colonies, the rest producing, after incubation for 96 h, small malformed colonies. Qualitatively, the same effect is observed when u.v.-irradiated wild-type M. radiodurans is incubated at 39 degrees C in the presence of nalidixic acid before plating. Compared with the loss of viability, the loss of normal colony development as a function of the radiation dose is sensitive, having I/e values of 210 ergs/mm2 for u.v. radiation and of 4 to 5 krad for 60Co gamma-radiation. These are identical to the radiation dose-response values of a recombination-deficient mutant of M. radiodurans. At first the abnormal colonies consist entirely of giant bacteria but eventually a few bacteria with normal morphology appear and because of their much faster generation time a highly sectored colony results. These colonies can be "rescued" by plating the irradiated bacteria held at 39 degrees C on agar containing pantoyl lactone, their growth being identical to that of unirradiated bacteria. Abnormal colony development is not a general phenomenon in temperature-sensitive mutants of M. radiodurans but occurs in those mutants which are sensitized to radiation when held at 39 degrees C. It is concluded that these abnormal colonies are produced as a result of a defect in a recombination function and that this function is also involved in the regulation of normal cell division.

Cell Division↗

Emendation of the genus Acidomonas Urakami, Tamaoka, Suzuki and Komagata 1989.

The genus Acidomonas and the species Acidomonas methanolica were recharacterized by using the type strain (NRIC 0498(T)), three reference strains and 10 methanol-utilizing bacteria that were isolated from activated sludge from three different sewage-treatment plants in Tokyo. Based on 16S rDNA sequences, all strains formed a single cluster within the Acetobacteraceae that was clearly different from the genera Acetobacter, Gluconobacter, Gluconacetobacter, Asaia and KOZAKIA: The 14 strains were identified as a single species, Acidomonas methanolica, by DNA-DNA similarities, showed DNA G+C contents that ranged from 62 to 63 mol% and had Q-10 as the major quinone, accounting for >87 % of total ubiquinones. Cells of Acidomonas methanolica had a single polar flagellum (or occasionally polar tuft flagella); this differs from a previous study that described peritrichous flagella. Oxidation of acetate was positive for all strains, but oxidation of lactate was weakly positive and varied with strains. Dihydroxyacetone was not produced from glycerol. Pantothenic acid was an essential requirement for growth. The strains tested grew at mostly the same extent at pH 3.0-8.0. Therefore, Acidomonas methanolica should be regarded as acidotolerant, not acidophilic. The descriptions of the genus Acidomonas and the species Acidomonas methanolica Urakami, Tamaoka, Suzuki and Komagata 1989 are emended with newly obtained data.

Acetobacteraceae↗

Neurospora mitochondria contain an acyl-carrier protein.

Mitochondria of Neurospora crassa were found to contain a protein which was labelled with [14C]pantothenic acid and which carried an acyl group. This protein, when purified 6000-fold, closely resembled the bacterial and chloroplast acyl-carrier protein(s) [ACP(s)] in its physical and chemical properties. The predominant acyl group esterified to the purified protein was 3-hydroxytetradecanoate, as determined by gas chromatographic mass spectrometry. The amino acid sequence of the tryptic peptide carrying the 4'-phosphophantetheine moiety showed a high degree of sequence similarity to the analogous bacterial and chloroplast ACP peptide sequences. The possible functions of this ACP in lipid metabolism are discussed in view of the fact that Neurospora has a separate cytoplasmic enzyme complex which carries out the de novo biosynthesis of fatty acids.

Acyl Carrier Protein↗

A kinetic study on pantetheinase inhibition by disulfides.

The mammalian enzyme pantetheinase, which hydrolyzes pantetheine to pantothenic acid and cysteamine, is inhibited by many thiol reagents and activated by thiols. Two thiol groups of different reactivity and accessibility are involved in the catalytic process [Ricci, G., Nardini, M., Chiaraluce, R., Duprè, S. & Cavallini, D. (1986) Biochim. Biophys. Acta 870, 82-91]. The inhibition kinetics by some natural and synthetic disulfides [pantethine, cystamine, 5,5'-dithiobis(2-nitrobenzoic acid), 4,4'-dithiodipyridine and oxidized mercaptoethanol] has been studied by two experimental approaches, either by monitoring activity after incubation of the enzyme with the inhibitor or by determining the progress curves in the presence of substrate and inhibitor. Data reported here indicate that pantetheinase reacts irreversibly with various disulfides in a time-dependent manner with the formation of a mixed disulfide apparently preceeded by a conformational change, giving a modified E* form with new kinetic parameters. This modified form may be further competitively inhibited by disulfides interacting with the enzyme at the active site.

Amidohydrolases↗

Nutritional requirements of Plasmodium falciparum in culture. II. Effects of antimetabolites in a semi-defined medium.

A semi-defined minimal medium, in which pantothenic acid is the only vitamin, was used to culture Plasmodium falciparum for the analysis of antimetabolite drugs. Analogs of riboflavin, nicotinamide, pyridoxine, and thiamin inhibited the growth of this parasite; for each drug, effects were much more pronounced after 96 h of exposure compared to 48 h. The most potent drug examined was 8-methylamino-8-desmethyl riboflavin (IC50 value approximately 1.0 X 10(-10) M at 96 h). Avidin, a protein which complexes and thus inactivates biotin, did not affect parasite viability. Other antimalarial drugs, including chloroquine and quinine derivatives and antibiotics, were equipotent in the minimal medium and in RPMI 1640. Four strains of P. falciparum showed only minor differences in sensitivity to these antimetabolites. The use of prolonged drug exposure times and a vitamin-depleted medium allowed the preliminary characterization of antimalarial antimetabolites in vitro.

Animals↗

The relationship between dietary intake and the number of teeth in elderly Japanese subjects.

OBJECTIVE: This study used a precise weighing method to assess whether tooth loss was related to nutrient intake in elderly Japanese subjects. MATERIAL AND METHODS: Fifty-seven subjects aged 74 years were randomly selected from a longitudinal interdisciplinary study of ageing. Complete 3-day food intake data were obtained by a precise weighing method. The dietary intakes of energy and nutrients were calculated based on the Standard Tables of Food Composition in Japan (5th ed.). A clinical evaluation of the number of teeth present was carried out. Multiple regression standardised coefficients for each nutrient was estimated based on a continuous scale adjusted for gender, smoking habits, and educational level. After dividing the subjects into two groups according to the number of teeth present (0-19, 20+), the difference in the intake of nutrients and the amount of food consumed per day was evaluated. RESULTS: The number of teeth present had a significant relationship with the intake of several nutrients. In particular, total protein, animal protein, sodium, vitamin D, vitamin B1, vitamin B6, niacin, and pantothenic acid were significantly associated with the number of teeth present and with the two groups (0-19, 20+). The intake of vegetables and fish, shellfish, and their products was significantly lower among subjects with fewer teeth. CONCLUSION: This study suggests that there was a significant relationship between nutrient intake, such as minerals and vitamins from food, and tooth loss.

Aged↗

Interactions of thiamin, riboflavin, and other B-vitamins.

Interactions of the B-complex vitamins are essential in the performance of metabolic and catabolic reactions in the body. Even vitamin C and the fat-soluble vitamins may be involved in these interactions. Clinical and biochemical aberrations associated with various disease states can often be explained on the basis of these vitamin interrelationships. Health and nutritional well-being are dependent upon the maintenance and proper functioning of these vitamin-dependent metabolic pathways.

Adult↗

Food preferences, dieting behaviors, and body image perceptions of elite figure skaters.

The objectives of this study were to determine food preferences, body image perceptions, dieting behaviors, and nutrient intakes of elite male and female figure skaters. Male (n = 23) and female (n = 26) figure skaters completed a food preference checklist, a questionnaire examining their demographics, dieting behaviors and body image perceptions, and 3-d food records. Male skaters had a higher preference (score > or = 6) for grains, fruit, meat, dairy, fats, and sweets. Female skaters had higher preference for grains and fruits. Of the female skaters, 30% considered themselves overweight and indicated a preference for a thinner body contour. Both male and female skaters expressed a preference for leaner body contours for members of the opposite gender. Total energy intake, total fat (females) and dietary fiber were below the dietary recommendations. Vitamin E, vitamin D, folate (females), pantothenic acid (females), calcium (females), magnesium, potassium, and phosphorus (females) were less than two-thirds of the dietary recommendations. Macronutrient intake of male skaters was associated with preferences for the grain group, although no association was observed among female skaters. Results suggest that these behaviors and attitudes need to be assessed and addressed among figure skaters, given their impact on dietary intakes and overall well-being.

Adult↗

The blood-brain barrier sodium-dependent multivitamin transporter: a molecular functional in vitro-in situ correlation.

The molecular mechanism of biotin brain uptake was investigated using an in vitro bovine blood-brain barrier (BBB) cell model and an in situ mouse brain perfusion technique. A functional uptake/transport correlation of the in vitro and in situ characteristics of biotin uptake was investigated. Morphological and immunochemical characteristics (e.g., factor VIII expression) of the primary culture of brain microvessel endothelial cells (BMECs) were confirmed. Gene expression of the multidrug resistance (Mdr1) and sodium-dependent multivitamin (SMVT) transporters was also determined in BMECs. Biotin transport was saturable and Na(+)-dependent at the luminal side of the BBB. The estimated half-saturation concentrations (K(m)) of biotin uptake in vitro and in situ were 49.1 and 35.5 microM, respectively, supporting the presence of a carrier-mediated biotin transport system. Inhibition studies using various biotin derivatives and structural analogs demonstrated the structural requirements for biotin-SMVT interaction. Desthiobiotin and pantothenic acid significantly inhibited the uptake of biotin, whereas 2-iminobiotin and diaminobiotin were very weak inhibitors. Based on our results, there was a good correlation between the in vitro and in situ BBB models, suggesting that when a single membrane transporter is involved in substrate uptake, flexibility in choosing the experimental model can be afforded. The current results are also consistent with the suggestion that the properties of the BBB are likely to be organ-specific rather than species-specific. Further mechanistic and comparative studies are needed to validate these results. In conclusion, the in vitro transporter-based mechanism studies produced valuable molecular functional transport results that correlated well with in situ results.

Animals↗

Availability of Essential B-Group Vitamins to Lactobacillus plantarum in Green Olive Fermentation Brines.

The availability throughout the traditional Spanish-style green olive fermentation of four vitamins that are essential for the growth of Lactobacillus plantarum was studied. It was found that nicotinic and pantothenic acids, biotin, and vitamin B(inf6) were available in the fermentation brines within the first few days of the process, and their levels throughout the fermentative process were well above those required by L. plantarum to grow at its maximum growth rate. In laboratory medium, various yeast strains isolated from the fermentations were found to produce these vitamins in amounts several times that required by L. plantarum. This finding suggests that some yeast species might play a role in encouraging the growth of L. plantarum in Spanish-style green olive fermentation.

Journal Article↗

Increased sensitivity of the microbiological assay for biotin by Lactobacillus plantarum.

Addition of Tween 80 to biotin assay medium containing acid-hydrolyzed casein as the amino acid source caused marked growth of Lactobacillus plantarum ATCC 8014 in the absence of added biotin. This growth-promoting activity could be eliminated by treating the "vitamin-free" Casamino Acids (Difco) with activated charcoal (Darco G-60) at pH 3.5 for 30 to 60 min. Incorporation of Tween 80 and charcoal-purified Casamino Acids (PCA) into the assay medium (0.8 g and 27 g, respectively, per liter of single strength medium) in place of unpurified Casamino Acids resulted in a medium in which L. plantarum responded to 30 to 50 times less biotin over an extended linear response range (1.3 logs versus 1.0 log) than was required for similar growth in the standard medium. Endogenous growth in the modified medium was absent if the inoculum used was of low density, if it was prepared from biotin-deficient cells, and if the reagents used were free from contaminating traces of biotin. Assays of biological materials for biotin content using the standard medium and the Tween 80-PCA-modified medium resulted in nearly identical values for all samples tested.

Biotin↗

Improvement of microbial assays of vitamins.

A method for the improvement of microbial assays of vitamins, which involves the addition of a surfactant to the incubated test, was developed. This surfactant tends to eliminate bacterial clumping, giving a uniform suspension of single cells, thereby making the turbidity readings less erratic and the actual assay standard curves more closely related to the desired theoretical curve.

Bacteriological Techniques↗

Transposon Tn5-induced mutagenesis of Rhizobium japonicum yielding a wide variety of mutants.

When the "suicide" vector pSUP1011, which carries transposon Tn5 (Kmr), was introduced into Rhizobium japonicum USDA 110, kanamycin-resistant (Kmr) colonies were detected at a frequency (4.2 X 10-6) ca. 30 times greater than the spontaneous kanamycin resistance frequency (1.4 X 10-7). Ten thousand Kmr mutants were isolated and tested for nutritional auxotrophy. Auxotrophs were detected at a frequency of 0.5%. The following classes of auxotrophs were identified: adenine- (three), histidine- (three), glutamate- (five), adenine plus thiamine- (nine), uracil- (three), pantothenic acid- (one), tryptophan- (three), and methionine- (three). Mutants blocked in symbiotic nitrogen fixation (Fix-) were also identified at a frequency of 3%. The glutamate auxotrophs were studied in more detail, and all five showed an altered expression of nitrogenase activity in free-living cultures.

Anti-Bacterial Agents↗

Nutrition of five Bacteroides strains.

Quinto, Grace (University of Kentucky College of Medicine, Lexington). Nutrition of five Bacteroides strains. J. Bacteriol. 84:559-562. 1962.-Some of the nutritional requirements of five gram-negative anaerobic bacilli, including Ristella perfoetens, Zuberella clostridiformis, and three Bacteroides strains freshly isolated from clinical exudates, were investigated. A fluid maintenance medium was developed in which the three freshly isolated strains, C-4, C-7, and C-2795, grew maximally in 12 to 24 hr. The maintenance medium contained 2.0% Trypticase and Proteose Peptone, 0.5% glucose, and 0.1% sodium thioglycolate; it was adjusted to pH 7.2 and supplemented with 0.1 mug of hemin/ml. Strains C-4, C-7, and C-2795 were cultivated through 14 serial cultures in fluid maintenance medium containing 0.1 mug of hemin/ml. The most satisfactory inoculum was a 1:100 or 1:1,000 dilution of a 24-hr seed culture. All the strains except Z. clostridiformis grew serially in a defined medium. R. perfoetens required pantothenic acid, nicotinic acid, and the following amino acids: histidine, tryptophan, tyrosine, valine, phenylalanine, cystine, and probably arginine, glutamic acid, methionine, glycine, isoleucine, leucine, and lysine. The C-4, C-7, and C-2795 strains required the hemin supplement in defined medium, but not vitamins, purines, or pyrimidines.

Amino Acids↗

Biosynthesis of diacetyl in bacteria and yeast.

Both diacetyl and acetoin were produced by cell-free extracts and cultures of Pseudomonas fluorescens, Aerobacter aerogenes, Lactobacillus brevis, and Saccharomyces cerevisiae 299, whereas only acetoin was produced by cell-free extracts and cultures of Streptococcus lactis, Serratia marcescens, Escherichia coli, and S. cerevisiae strains 513 and 522. Cell-free extracts that produced diacetyl did not produce it from acetoin; they produced it from pyruvate, but only if acetyl-coenzyme A was was added to the reaction mixtures. Production of diacetyl by S. cerevisiae 299 was prevented by valine, inhibited by sodium arsenite, and stimulated by pantothenic acid. Valine did not prevent the production of acetoin. E. coli and the three strains of S. cerevisiae did not decarboxylate alpha-acetolactate but did use acetaldehyde in the production of acetoin from pyruvate. The other organisms produced acetoin from pyruvate via alpha-acetolactate.

Bacteria↗

Vanin-1-/- mice exhibit a glutathione-mediated tissue resistance to oxidative stress.

Vanin-1 is an epithelial ectoenzyme with pantetheinase activity and generating the amino-thiol cysteamine through the metabolism of pantothenic acid (vitamin B(5)). Here we show that Vanin-1(-/-) mice, which lack cysteamine in tissues, exhibit resistance to oxidative injury induced by whole-body gamma-irradiation or paraquat. This protection is correlated with reduced apoptosis and inflammation and is reversed by treating mutant animals with cystamine. The better tolerance of the Vanin-1(-/-) mice is associated with an enhanced gamma-glutamylcysteine synthetase activity in liver, probably due to the absence of cysteamine and leading to elevated stores of glutathione (GSH), the most potent cellular antioxidant. Consequently, Vanin-1(-/-) mice maintain a more reducing environment in tissue after exposure to irradiation. In normal mice, we found a stress-induced biphasic expression of Vanin-1 regulated via antioxidant response elements in its promoter region. This process should finely tune the redox environment and thus change an early inflammatory process into a late tissue repair process. We propose Vanin-1 as a key molecule to regulate the GSH-dependent response to oxidative injury in tissue at the epithelial level. Therefore, Vanin/pantetheinase inhibitors could be useful for treatment of damage due to irradiation and pro-oxidant inducers.

Amidohydrolases↗