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Vitamin K-dependent carboxylation. Evidence that at least two microsomal dehydrogenases reduce vitamin K1 to support carboxylation.

It has been shown that NAD(P)H dehydrogenase (EC 1.6.99.2) reduces vitamin K1 and can support the [vitamin K1 + NADH]-dependent carboxylation reaction in rat liver microsomes (Wallin, R., Gebhardt, O., and Prydz, H. (1978) Biochem. J. 169, 95-101). Antibodies were raised in rabbits against the purified enzyme from liver cytosol and used to study the importance of NAD(P)H dehydrogenase in the vitamin K-dependent carboxylation reaction. The antibodies neutralized the warfarin-sensitive NAD(P)H dehydrogenase activity in Triton X-100-solubilized microsomes; however, they neutralized only 45% of the total [vitamin K1 + NADH]-dependent carboxylation activity. Chromatography on protein A-sepharose showed that the remaining carboxylase activity was not the result of soluble antigen-antibody complexes. The data presented support the conclusion that the microsomal preparation also contains a non-warfarin-sensitive dehydrogenase(s) that, in addition to NAD(P)H dehydrogenase, can reduce vitamin K1 to support the carboxylation reaction.

Animals↗

Incidence of squamous cell carcinoma in hairless mice irradiated with ultraviolet light in relation to intake of ascorbic acid (vitamin C) and of D, L-alpha-tocopheryl acetate (vitamin E).

We have carried out a study of large malignant skin tumours (squamous-cell carcinomas) and other lesions in "hairless" mice (in groups of 45 or 60 mice) intermittently exposed to ultraviolet light over a period of 15 weeks, beginning when the mice were about 8 weeks old. Various groups were given a standard diet (Wayne Lab-Blox) or the same food with added vitamin C or vitamin E throughout the study. Lesions, classified by histopathologic study as atypical squamous-cell proliferations varying from early actinic keratoses to invasive poorly differentiated squamous-cell carcinomas, had begun to develop by the end of the period of irradiation. They were counted twice a month for five months. The observed fraction of mice that developed lesions during successive time periods was analyzed by the statistical method recommended by a committee of the International Agency for Research on Cancer. A pronounced effect of vitamin C in decreasing the incidence of the malignant lesions was observed with very high statistical significance. No significant effect of vitamin E was observed. We conclude that vitamin C should be given special attention with respect to the relation between diet and cancer.

Animals↗

Analysis of fat-soluble vitamins. XXVII. High performance liquid chromatographic and gas-liquid chromatographic determination of vitamin D in fortified milk and milk powder: collaborative study.

A collaborative study of the high performance liquid chromatographic (HPLC) method for vitamin D in fortified milkpowder (skimmed and whole milk) and a milkpowder preparation was carried out on 182 samples distributed to 26 laboratories. Thirteen laboratories submitted results. The level of vitamin D was 2-7 IU vitamin D3/g milkpowder. All samples also contained vitamin A. Three laboratories were excluded from the statistical evaluation because of incomplete results or deviation from the analytical procedure. Other laboratories reported acceptable results. At the same time, 63 samples were distributed to 9 laboratories which used a gas-liquid chromatographic method for determining vitamin D in milkpowder. Only one laboratory reported results. The HPLC method has been adopted official first action.

Animals↗

Effects of dietary vitamin D intake on plasma levels of parathyroid hormone and vitamin D metabolites in healthy Japanese.

To clarify the nutritional status of vitamin D in Japanese, effects of dietary intake of vitamin D on plasma levels of intact and highly sensitive parathyroid hormone (I-PTH and HS-PTH), 25-hydroxyvitamin D (25-OH-D), 1,25-dihydroxyvitamin D (1,25(OH)2D), calcium (Ca) and phosphorus (P(i)) in 79 healthy Japanese were investigated. The plasma levels of 25-OH-D in men were significantly higher than those in women, whereas those of HS-PTH in men were significantly lower than those in women. The levels of 25-OH-D in men were generally higher than those in women. Significant correlations were observed between the dietary vitamin D intake and the plasma 25-OH-D or HS-PTH levels. Correlations between the plasma 25-OH-D levels and the plasma HS-PTH levels were also significant. These results suggest that dietary intake of sufficient amounts of vitamin D is effective for improving the vitamin D nutritional status through normalizing PTH levels.

Adult↗

Influence of antimicrobial chemotherapy and smoking status on the plasma concentrations of vitamin C, vitamin E, beta-carotene, acute phase reactants, iron and lipid peroxides in patients with pulmonary tuberculosis.

SETTING: Inflammation-related oxidative stress has been implicated in the pathogenesis of lung fibrosis and dysfunction in patients with pulmonary tuberculosis. OBJECTIVE: To investigate the effects of antimicrobial chemotherapy and smoking status on the plasma concentrations of the anti-oxidative nutrients vitamin C, vitamin E and beta-carotene, as well as those of iron, lipid peroxides and the acute phase reactants C-reactive protein (CRP) and ferritin. DESIGN: A total of 41 patients with active pulmonary tuberculosis were studied at the outset and after 6 months of antimicrobial chemotherapy. RESULTS: Initial plasma concentrations of vitamin C and beta-carotene were low, returning to normal values after chemotherapy in the non-smokers, but not in the smokers, while those of vitamin E remained low throughout in both groups. Ferritin and CRP concentrations decreased significantly following chemotherapy, with the former higher in smokers than in non-smokers. Serum lipid peroxides were elevated in patients with pulmonary tuberculosis and were unaffected by chemotherapy or smoking habits, while iron levels were not significantly affected by chemotherapy. Although residual dysfunction and infiltration were evident, pulmonary function (FEV1) and radiographic score improved equally in both smokers and non-smokers following antimicrobial chemotherapy. CONCLUSIONS: Even after 6 months of apparently successful antimicrobial chemotherapy, pulmonary tuberculosis is associated with increased oxidative stress, which is unrelated to cigarette smoking and characterized by increased levels of circulating lipid peroxides and low concentrations of plasma vitamin E.

Acute-Phase Proteins↗

Simultaneous determination of vitamins A, E and 25-OH-vitamin D: application in clinical assessments.

OBJECTIVE: To optimize an HPLC method for the simultaneous determination of vitamins A, E and 25-OH-vitamin D. METHODS: Modifications include sample volume, internal standard, temperature, detection and injection volume. RESULTS AND DISCUSSION: Validity was supported by performance in interlaboratory exercises. Utility and versatility in clinical practice were assessed in outpatients and hospitalized subjects. The method was shown to be accurate and reliable for the simultaneous determination of vitamins A, E and 25-OH-vitamin D in serum throughout the entire range of clinically relevant concentrations.

Chromatography, High Pressure Liquid↗

The role of universal distribution of vitamin A capsules in combatting vitamin A deficiency in Bangladesh.

Vitamin A deficiency is a major public health problem among preschool-aged children in many developing countries. In Bangladesh, a national nutritional surveillance system was initiated in 1990 to monitor 1) the occurrence of vitamin A deficiency by history of night blindness and 2) the routine coverage of national twice-yearly prophylactic vitamin A capsule (VAC) distribution. This study comprised data collected from June 1990 to August 1994. The VAC distribution had a mean coverage rate of 48.7% (95% confidence interval (CI) 48.4-49.0) in the rural areas; the coverage rate in the urban slums was 93.7% (95% CI 93.4-94.0). In the rural areas, the mean prevalence of night blindness was 0.86% (95% CI 0.81-0.91) and the bimonthly prevalence of night blindness ranged from 0.50% (95% CI 0.32-0.77) to 1.48% (95% CI 1.19-1.85), while in the urban slums the mean prevalence was 0.22% (95% CI 0.18-0.28) and the bimonthly prevalence ranged from zero to 0.62% (95% CI 0.27-1.37). The efficiency of VAC distribution was 27% (95% CI 17.6-35.3) in the rural areas and 77.8% (95% CI 61.8-87.1) in the urban slums. After adjustment for multiple potentially confounding factors, VAC receipt by individual children reduced the risk of night blindness in both rural and urban areas (rural areas: odds ratio (OR) = 0.74, 95% CI 0.63-0.87; urban slums: OR = 0.39, 95% CI 0.19-0.82). Breastfeeding was a protective factor for night blindness in both rural (OR = 0.53, 95% CI 0.42-0.67) and urban (OR = 0.32, 95% CI 0.15-0.66) areas. Night blindness was inversely related to the level of routinely attained coverage, and the degree of protection was associated with the time interval between the moment of VAC receipt and the moment of data collection. Although the prevalence of vitamin A deficiency in Bangladesh has been considerably lower in the 1990s than it was in the 1980s, it is still prevalent at all socioeconomic levels. Supplementation with high-dose VACs is an effective strategy for reducing night blindness, but the efficiency of the program will improve when coverage in the rural areas increases.

Bangladesh↗

Effect of 26,26,26,27,27,27-Hexafluoro-1,25-dihydroxyvitamin D3 on the expression of vitamin-D-responsive genes in vitamin-D-deficient mice.

26,26,26,27,27,27-Hexafluoro-1,25-dihydroxyvitamin D3 (ST-630) is a newly developed agent to maintain the levels of calcium and phosphorus in blood. Herein, we investigated the effect of this compound on the expression of vitamin-D-responsive genes in vitamin-D-deficient mice. ST-630 was more effective than 1, 25-dihydroxyvitamin D3 [1,25(OH)2D3] with respect to the induction of Cyp24 and calbindin-D9k mRNAs in the kidney and in the small intestine. Moreover, the increase in mRNA levels of vitamin-D-responsive genes induced by ST-630 lasted longer than that induced by 1,25(OH)2D3. These results indicate that ST-630 was more effective in inducing Cyp24 and calbindin-D9k gene expression than 1, 25(OH)2D3 when both compounds were injected into vitamin-D-deficient mice.

Animals↗

Evaluation of a pediatric multiple vitamin preparation for total parenteral nutrition. II. Blood levels of vitamins A, D, and E.

This study represents the first attempt to evaluate the American Medical Association Nutrition Advisory Group (NAG) recommendations for intravenous vitamin A, D, and E dosages for infants and children. Patients studied included 18 preterm infants (group 1) and 26 term infants and children (group 2A) receiving total parenteral nutrition for 2 to 4 weeks and eight infants and children receiving total parenteral nutrition for 3 to 6 months (group 2B). Term gestation infants and children up to 11 years of age all received the same dosages (those that were recommended by the NAG for children weighing more than 10 kg). Preterm infants received 65% of these doses. In group 1, cord blood alpha-tocopherol levels were less than 0.22 mg/dL in seven preterm infants (reference value = 0.29 +/- 0.04), but mean levels increased to 1.65 +/- 0.17 mg/dL after four days of treatment. Eight infants consistently received additional vitamin E orally (80 to 150 mg daily), and their levels increased to 2.18 +/- 0.26 mg/dL by four days of study and to 3.49 +/- 0.57 mg/dL after 3 weeks. Oral supplementation in the preterm infants appeared to be unnecessary because intravenous vitamins alone maintained levels above 1.1 mg/dL. In group 2, alpha-tocopherol levels were maintained within the reference range. Patients receiving lipid emulsions containing substantial quantities of alpha-tocopherol had significantly higher blood levels than patients receiving lipid emulsions containing little alpha-tocopherol (P less than .01). Mean 25-OH vitamin D levels were maintained above or within the reference range in groups 2A and 2B.(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗

[Neonatal vitamin K prophylaxis and vitamin k deficiency hemorrhages in Switzerland 1986-1988].

On the basis of an inquiry held in Switzerland in May 1988, over 99% of all newborn receive vitamin K prophylaxis, 59% orally and 41% intramuscularly. In the previous 2 1/2 years, ten cases of bleeding due to vitamin K deficiency had been observed, of which two were inadequately documented. In two children there was early haemorrhage and late haemorrhage in eight. The latter were all exclusively breast-fed and had received oral vitamin K prophylaxis. Seven presented with vitamin K deficiency due to cholestasis or chronic diarrhea. The only "idiopathic" case is insufficiently documented. The advantages and disadvantages of oral and intramuscular prophylaxis are discussed. A definite stand in favour of the one route or the other is not possible at present. However, the continuation of general prophylaxis is undoubtedly necessary.

Female↗

Vitamine--vitamin. The early years of discovery.

In 1905, Cornelius Adrianus Pekelharing found that animals fed purified proteins, carbohydrates, fats, inorganic salts, and water would thrive only if small amounts of milk were added to the diet. He concluded that the milk contained some unrecognized substance that in very small quantities was necessary for normal growth and maintenance. In 1911, Casimir Funk isolated a concentrate from rice polishings that cured polyneuritis in pigeons. He named the concentrate "vitamine" because it appeared to be vital to life and because it was probably an amine. Although the concentrate and other "accessory food substances" were not amines, the name stuck, but the final "e" was dropped. In 1913 two groups discovered a "fat-soluble" accessory food substance. Initially believed to be a single vitamin, two separate factors were involved. One, effective against xerophthalmia, was named vitamin A; the other, effective against rickets, was named vitamin D. The factor that prevented scurvy was isolated in 1928. Known as "water-soluble C," it was renamed ascorbic acid.

Ascorbic Acid↗

Growth control of human colon-adenocarcinoma-derived Caco-2 cells by vitamin-D compounds and extracellular calcium in vitro: relation to c-myc-oncogene and vitamin-D-receptor expression.

The human colon-cancer cell line Caco-2, though of malignant origin, is still able to express the c-myc proto-oncogene in a regulable fashion. Transition from the logarithmic growth phase into the quiescent, i.e., confluent state, is accompanied by a significant increase in the number of cells in the G0/G1 phase of the cell cycle and a concomitant reduction of c-myc mRNA and of nuclear association of c-myc protein. Conversely, growth stimulation by lowering extracellular [Ca++]0 to 0.25 mM results in up-regulation of c-myc expression levels and consequently inhibition of re-entry of Caco-2 cells into the G0/G1 phase. In contrast, regulation of c-myc in Caco-2 cells is completely resistant to vitamin-D sterols, since the anti-mitogenic action of I alpha, 25-dihdroxyvitamin D3 (I alpha, 25(OH)2D3) and of 2 synthetic analogs, I alpha, 25(OH)2-16-ene-23-yne-D3 and I alpha, 25(OH)2-26, 27-F6-16-ene-23-yne-D3, occurred independently of any change in c-myc mRNA and nuclear protein levels. Although the antiproliferative effect of the vitamin-D sterols requires high-affinity binding to the cytoplasmic vitamin-D receptor (VDR), vitamin-D sterols have no effect on VDR mRNA levels in Caco-2 cells. However, VDR mRNA expression changed in an antiparallel fashion to c-myc regulation upon transition between different growth states. This suggests that VDR mRNA abundance could nevertheless be important for vitamin-D-related c-myc-independent growth control in Caco-2 cells.

Antineoplastic Agents↗

1alpha,25-dihydroxy-16-ene-23-yne-vitamin D3 and 1alpha,25-dihydroxy-16-ene-23-yne-20-epi-vitamin D3: analogs of 1alpha,25-dihydroxyvitamin D3 that resist metabolism through the C-24 oxidation pathway are metabolized through the C-3 epimerization pathway.

The secosteroid hormone 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] is metabolized in its target tissues through modifications of both the side chain and the A-ring. The C-24 oxidation pathway, the previously well established main side chain modification pathway, is initiated by hydroxylation at C-24 of the side chain. The C-3 epimerization pathway, the newly discovered A-ring modification pathway, is initiated by epimerization of the hydroxyl group at C-3 of the A-ring. The end products of the metabolism of 1alpha,25(OH)2D3 through the C-24 oxidation and the C-3 epimerization pathways are calcitroic acid and 1alpha,25-dihydroxy-3-epi-vitamin-D3 respectively. During the past two decades, numerous noncalcemic analogs of 1alpha,25(OH)2D3 were synthesized. Several of the analogs have altered side chain structures and as a result some of these analogs have been shown to resist their metabolism through side chain modifications. For example, two of the analogs, namely, 1alpha,25-dihydroxy-16-ene-23-yne-vitamin D3 [1alpha,25(OH)2-16-ene-23-yne-D3] and 1alpha,25-dihydroxy-16-ene-23-yne-20-epi-vitamin D3 [1alpha,25(OH)2-16-ene-23-yne-20-epi-D3], have been shown to resist their metabolism through the C-24 oxidation pathway. However, the possibility of the metabolism of these two analogs through the C-3 epimerization pathway has not been studied. Therefore, in our present study, we investigated the metabolism of these two analogs in rat osteosarcoma cells (UMR 106) which are known to express the C-3 epimerization pathway. The results of our study indicate that both analogs [1alpha,25(OH)2-16-ene-23-yne-D3 and 1alpha,25(OH)2-16-ene-23-yne-20-epi-D3] are metabolized through the C-3 epimerization pathway in UMR 106 cells. The identity of the C-3 epimer of 1alpha,25(OH)2-16-ene-23-yne-D3 [1alpha,25(OH)2-16-ene-23-yne-3-epi-D3] was confirmed by GC/MS analysis and its comigration with synthetic 1alpha,25(OH)2-16-ene-23-yne-3-epi-D3 on both straight and reverse-phase HPLC systems. The identity of the C-3 epimer of 1alpha,25(OH)2-16-ene-23-yne-20-epi-D3 [1alpha,25(OH)2-16-ene-23-yne-20-epi-3-epi-D3] was confirmed by GC/MS and 1H NMR analysis. Thus, we indicate that vitamin D analogs which resist their metabolism through the C-24 oxidation pathway, have the potential to be metabolized through the C-3 epimerization pathway. In our present study, we also noted that the rate of C-3 epimerization of 1alpha,25(OH)2-16-ene-23-yne-20-epi-D3 is about 10 times greater than the rate of C-3 epimerization of 1alpha,25(OH)2-16-ene-23-yne-D3. Thus, we indicate for the first time that certain structural modifications of the side chain such as 20-epi modification can alter significantly the rate of C-3 epimerization of vitamin D compounds.

Animals↗

Autoradiographic studies with 3H 1,25 (OH)2 vitamin D2 and 3H 25 (OH) vitamin D3 in rat parathyroid glands.

After injection of radiolabeled 1,35 (OH)s vitamin D3, nuclear concentration of radioactivity is observed in parenchymal cells o hte parathyroid gland in pregnant, adult male, and 10-day male neonatal rats. In competition studies with unlabeled 1,25 (OH)2 vitamin D3, but not with 25 (OH) vitamin D3, in contrast to 3H 1,25 (OH)2 vitamin D3, do not show nuclear concentration in cells of te parathyroid. The results of te autoradiographic studies suggest the presence of receptors for a direct effect of 1,35 (OH)2 vitamin D3 on the parathyroid gland for modulation of parathyroid hormone secretion.

Aging↗

Vitamin B2-enhancement of sodium chromate (VI)--Induced DNA single strand breaks: ESR study of the action of vitamin B2.

Incubation of Chinese hamster V-79 cells with Na2CrO4 plus vitamin B2 resulted in an increase of Na2CrO4-induced DNA single strand breaks. Electron spin resonance (ESR) studies showed that vitamin B2 enhanced the formation of both hydroxyl radical and tetraperoxochromate (V) during the reaction of Na2CrO4 with hydrogen peroxide. Furthermore, ESR studies demonstrated that a chromium (V) species with a g value of 1.977 was formed by the reaction of Na2CrO4 with vitamin B2. These results indicate that chromate reacts with vitamin B2 to form chromium (V) species and also suggest that the enhancement effect of vitamin B2 on chromate-induced DNA single strand breaks may result from an increase of chromium (V)-related hydroxyl radical formation.

Animals↗

Effects of 1alpha-hydroxy-vitamin D3 and 1,25-dihydroxy-vitamin D3 on calcium and phosphorus metabolism in hypoparathyroidism.

The effects of 1alpha-OH D3 or 1,25-(OH)2D3 on calcium and phosphorus metabolism have been evaluated in five hypoparathyroid patients to establish the direct effects of these compounds in adult humans, uncomplicated by compensatory changes in parathyroid hormone secretion. Doses of 1-2.5 mug/day in four patients (5 mug/day in a fifth patient on diphenylhydantoin and phenobarbital) caused a marked increase in serum calcium concentration and urinary calcium excretion, without significant changes in renal calcium clearance or urinary hydroxyproline excretion. These results suggest that the correction of hypocalcemia involved primarily a stimulation of intestinal calcium absorption rather than a stimulation of skeletal calcium resorption. Simultaneously, there were increases in urinary phosphorus excretion and variable changes in serum inorganic phosphate concentration. These effects were produced by doses of 1alpha-OH D3 and 1,25-(OH)2D3 which approach the dose needed to prevent rickets, in contrast to the very large doses of vitamin D or 25-OH D3 required for comparable effects in hypoparathyroid patients. The increased relative effectiveness of these one-hydroxylated forms of vitamin D reveals a deficiency of vitamin D one-hydroxylation in hypoparathyroidism. The rapidity of action of 1alpha-OH D3 and 1,25-(OH)2D3 was also striking. Apart from its physiologic implications, the potency of the one-hydroxylated forms of vitamin D offers significant therapeutic advantages in some patients whose hypoparathyroidism is difficult to control with vitamin D itself.

Adult↗

Vitamin D3 polyunsaturated side-chain analogues (EB1089, GS1590) and the 20-epi-vitamin D3 analogue CB1393 suppress parathyroid hormone secretion and mRNA level in bovine parathyroid cells.

Several vitamin D analogues, with reduced hypercalcemic and hyperphosphatemic toxicity at therapeutic dosages, are in clinical use for prevention and treatment of secondary hyperparathyroidism (HPT) in chronic renal failure. We have performed a first in vitro evaluation of five vitamin D analogues displaying less calcemic activity in normal rats, considerably more antiproliferative ability and higher transcription activation potential than 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), with the future prospects in mind to identify even more effective and less calcemic vitamin D analogues for treatment of HPT. The vitamin D analogues EB1089 and GS1590 have polyunsaturated side-chains, whereas HEP187, MC1598 and CB1393 display altered stereochemistry at carbon 20. In primary cultures of bovine parathyroid cells EB1089, GS1590, CB1393 and MC1598 as well as the comparative controls 1,25(OH)(2)D(3), 22-oxacalcitriol (OCT, maxacalcitol), 19-nor-1,25(OH)(2)D(2) (paricalcitol) and 1alpha(OH)D(2) (doxercalciferol) significantly suppressed PTH secretion or reduced PTH mRNA level at 10(-8), 10(-10), and 10(-11)M for all compounds except for MC1598 at the lowest concentration. The analogue HEP187 displayed no PTH suppressive activity. We conclude that the vitamin D analogues EB1089, GS1590 and CB1393 may be suitable for treatment of hyperparathyroidism secondary to uremia and that further evaluation in vivo should be considered.

Animals↗

Chromatographic separation of vitamin D3 sulfate and vitamin D3.

This paper describes a simple chromatographic technique on Sephadex LH20 for the separation of vitamin D3 sulfate from free vitamin D3 and its metabolites. This technique has been used in the study of vitamin D3 sulfate metabolism in rats. Seven hours after injection of vitamin D3 sulfate (35S or 35S and 3H) only the peak of vitamin D sulfoconjugate was found in chromatographic elution of serum extracts.

Animals↗