Vitamin K-deficient Diet Research Articles

Effect of latamoxef on platelet function and prothrombin time in partially nephrectomized rats.

Latamoxef (daily 100 and 300 mg/kg, i.v.) was injected once a day for 8 days to 75% and 90% nephrectomized rats kept on a vitamin K-sufficient diet (500-600 ng/g) or a vitamin K-deficient diet (30-50 ng/g), and changes in ADP-induced platelet aggregation and prothrombin time were examined. The half-life of latamoxef was markedly prolonged and plasma latamoxef and N-methyltetrazolethiol (NMTT) concentrations increased, resulting in a delay of the total body clearance of the compounds. The ADP-induced platelet aggregation increased after nephrectomy, and latamoxef slightly but inconsistently decreased the aggregation. Prothrombin time did not change even in the 90% nephrectomized rats kept on an ordinary diet, but increased dose-dependently in the vitamin K-deficient nephrectomized rats, with the 90% nephrectomized animals showing larger increases of prothrombin time. These data suggest that NMTT or NMTT-containing antibiotics cause no hypoprothrombinemia even in partially nephrectomized rats when they are fed an ordinary diet containing vitamin K, but these compounds enhance the manifestation of hypoprothrombinemia in vitamin K deficiency. Further renal failure promotes the manifestation by increasing drug concentration in the blood. However, platelet aggregation in these animals is not significantly affected at the doses examined.

Effects of Antibiotics on Fibrinolytic Activity In Vivo

The effects of latamoxef, cefamandole, carbenicillin and cefotaxime on fibrinolytic system in rats were examined under feeding of an ordinary diet or a vitamin K-deficient diet for eight days. No obvious change was observed in UK-induced plasma ciot lysis time, euglobulin lysis time and antiplasmin level in plasma. These findings of the in vitro and ex vivo studies suggest that these antibiotics cause no enhancement of fibrinolytic activity.

Effects of antibiotics on fibrinolytic activity in vivo.

The effects of latamoxef, cefamandole, carbenicillin and cefotaxime on fibrinolytic system in rats were examined under feeding of an ordinary diet or a vitamin K-deficient diet for eight days. No obvious change was observed in UK-induced plasma clot lysis time, euglobulin lysis time and antiplasmin level in plasma. These findings of the in vitro and ex vivo studies suggest that these antibiotics cause no enhancement of fibrinolytic activity.

Liver and Plasma Levels of Descarboxyprothrombin (PIVKA II) in Vitamin K Deficiency in Rats

Descarboxyprothrombin (PIVKA II) is a precursor of prothrombin without biological activity, and it increases with vitamin K deficiency. We studied the time course changes in liver and plasma levels of PIVKA II during the progress of vitamin K deficiency in rats. Good correlation was observed between liver PIVKA II and plasma PIVKA II and between liver or plasma PIVKA II and plasma prothrombin in experiments In which rats were fed a vitamin K-deficlent diet. Feeding of a vitamin K-deficient diet or fasting caused marked Increases in liver and plasma PIVKA II In male rats and a weaker response in female rats. Warfarin, a vitamin K antagonist, caused an abrupt increase in liver PIVKA II, but the Increase In plasma PIVKA II was delayed about 3 hr. Plasma prothrombin decreased from about 30 min later. Factor VII decreased similarly to prothrombin, and changes in the prothrombin time and activated partial thromboplastin time were slower than the changes in these substances. Sex differences were not seen in these warfarin actions. These observations indicate that liver and plasma PIVKA II are sensitive markers of vitamin K deficiency in rats, and assay of PIVKA II can be useful for analyzing the action mechanism of drugs which Influence blood coagulation.

Production of hypoprothrombinaemia by cefazolin and 2-methyl-1,3,4-thiadiazole-5-thiol in the rat.

Recent findings have suggested that the 1-methyltetrazole-5-thiol (MTT) group contained in several beta-lactam antibiotics may be responsible for the hypoprothrombinaemia associated with these drugs. In order to determine if the hypoprothrombinaemia associated with cefazolin is due to the presence of the structurally related 2-methyl-1,3,4,-thiadiazole-5-thiol (MTD) group which it possesses, the ability of MTD and cefazolin to produce hypoprothrombinaemia in rats was examined. Female rats maintained on a vitamin K-deficient diet for ten days developed hypoprothrombinaemia after the intravenous administration of cefazolin for two subsequent days. Hypoprothrombinaemia was also produced by the oral administration to vitamin K-deficient rats of either cefazolin or MTD, while the oral administration of cefotaxime, which does not contain a thiol group, had no effect. In a rat liver microsomal system, MTD was found to be a much more potent inhibitor than cefazolin of the vitamin K-dependent step in clotting factor synthesis, the gamma-carboxylation of glutamic acid. These results suggest that the hypoprothrombinaemia associated with cefazolin may be due to the MTD group.

Coagulopoietin activity in rat plasma: Influence of degree of hypoprothrombinemia and reproducibility of the response

Injection of plasma from a vitamin K-deficient hypoprothrombinemic rat into a normal rat causes an increase in plasma prothrombin activity in the recipient rat. Maximum concentrations of the factor responsible for this response (coagulopoietin) are present in the plasma of rats fed a vitamin K-deficient diet for 7 days and do not increase as the period of deficiency is increased. The coagulopoietin activity of plasma from individual rats is variable, but similar activity was observed when multiple recipient rats received the same donor plasma.

24] γ-carboxyglutamic acid

Publisher Summary This chapter provides an overview of γ-carboxyglutamic acid. In vitro synthesis of 4-carboxyglutamic acid (Gla) residues can be carried out using microsomes, which have accumulated endogenous precursor protein, as a result of treatment of the animal with either a vitamin K-deficient diet or warfarin. When detergent-solubilized microsomes are utilized, synthesis of Gla can be demonstrated with synthetic peptide substrates, homologous to short sequences in prothrombin, one of which is Phe-Leu-Glu-Glu-Leu (FLEEL), the sequence of residues 5–9 in rat prothrombin. To measure Gla synthesis in endogenous accumulated substrate in vitro, it is necessary to prepare microsomes from vitamin K-deficient or sodium warfarin-treated animals. Several methods exist for the direct analysis of Gla in protein hydrolysates. These methods employ automated amino acid analysis, or high-pressure liquid chromatography, or manual ion-exchange chromatography followed by fluorescent or absorption spectroscopy.

Production of hypoprothrombinemia by moxalactam and 1-methyl-5-thiotetrazole in rats.

To determine whether the hypoprothrombinemia associated with antibiotics containing a 1-methyl-5-thiotetrazole (MTT) group is a result of the presence of the MTT group, rats were maintained on a vitamin K-deficient diet for 10 days and then received either intravenous moxalactam or cefotaxime or oral MTT for two additional days. MTT and moxalactam, which contains the MTT group, prolonged prothrombin time. Cefotaxime, which lacks the MTT group, had no effect.

Vitamin K-dependent carboxylation of glutamic acid residues to γ-carboxyglutamic acid in lung microsomes

Vitamin K-dependent carboxylation of glutamic acid residues to γ-carboxyglutamic acid was demonstrated in proteins of lung microsomes. The carboxylation was 12% of that in liver microsomes per milligram of mierosomal protein. Carboxylation was very low with microsomes of untreated rats but increased with time up to 42 h after warfarin administration. Carboxylation was highest with microsomes from rats fed a vitamin K-deficient diet. This suggests that a protein(s) accumulates which can be carboxylated in vitro/J. Lung microsomes also catalyzed the vitamin K-dependent carboxylation of the peptide Phe-Leu-Glu-Glu-Leu. The peptide carboxylase activity was 9% of that obtained with liver microsomes. Vitamin K-dependent protein carboxylation required NADH or dithioerythritol, suggesting that vitamin K had to be reduced to the hydroquinone. Accordingly, vitamin K 1 hydroquinone had carboxylating activity without added reducing agents. Menaquinone-3 was considerably more active than phylloquinone. The temperature optimum for carboxylation was around 27 °C.

Aortic glycosaminoglycans in avitaminosis

In hypovitaminosis K induced by a vitamin K-deficient diet or by Pelentan (ethyl biscoumacetate) a decrease was observed in the total content of glycosaminoglycans (GAG) and a change in the relative proportions of their fractions in the aortic wall of rats. Daily administration of ascorbic acid (100 mg/kg) to the rats from the first day of their vitamin K-deficient diet prevented changes in sulfated GAG but had no effect on the decrease in the hyaluronic acid level. The results are discussed from the standpoint of the disturbance of ascorbic acid metabolism in hypovitaminosis K.

Vitamin K-dependent carboxylase: Increased activity in a hypoprothrombinemia state

Rat liver microsomes contain a vitamin K-dependent carboxylase activity that converts protein-bound glutamyl residues to γ-carboxy glutamyl residues. This activity has been studied utilizing low molecular weight peptides (peptide carboxylase) and endogenous microsomal precursor proteins (protein carboxylase) as substrates for the enzyme. Feeding rats a vitamin K-deficient diet or the administration of Warfarin, causes a 2- to 3-fold increase in these activities. The increase in peptide carboxylase activity is not dependent on the presence of increased amounts of prothrombin precursors in the microsomes and is more closely correlated with the degree of plasma hypoprothrombinemia than with the level of these precursors in the liver. The protein carboxylase activity appears to reflect the level of precursor substrates in the liver. The apparent induction of peptide carboxylase activity is not due to an alteration in affinity of the enzyme for its substrate. An apparent rapid turnover of the enzyme complicated the interpretation of experiments which utilized the protein synthesis inhibitor cycloheximide. The data were, however, consistent with the hypothesis that the increased activity measured represents a synthesis of an increased amount of enzyme rather than an activation of existing inhibited activity.

Estrogen and prothrombin synthesis: effect of estrogen on absorption of vitamin K1.

Intact male and female albino rats fed a vitamin K-deficient diet develop a plasma prothrombin-proconvertin deficiency. Male rats respond with a precipitous drop to approximately 20-30% of normal plasma levels within 2-5 days, whereas female rats respond at a slower rate. Ethynylestradiol, 5-10 mug/day, or castration, reduces the progressive decline of plasma prothrombin-proconvertin seen in nonsupplemented intact male rats. The response of castrate females differs little from the response of intact females. Ethynylestradiol, 5-10 mug/day, affects both castrate males and females similarly, limiting the prothrombin-proconvertin decrease to about 13% below control value after 14 days. Intestinal absorption of vitamin K1 measured in the thoracic duct lymph of pentobarbital-anesthetized castrate male and female rats was shown to increase significantly after estrogen treatment. Estrogen-treated castrate male and female rats absorbed 25.8 mug and 11.8 mug vitamin K1, respectively. Nontreated control castrate male and female rats absorbed 0.0 mug and 1.2 mug, respectively, during a 240-min collection period. Use of radioactive vitamin K1 in similar experiments confirmed these results. Estrogen-treated castrate males absorbed vitamin K1 at the rate of 30-40 mug/g lymph whereas nontreated control males absorbed only about 6 mug/g lymph.

Effects of Vitamin K Deficiency, Warfarin, and Inhibitors of Protein Synthesis upon the Plasma Levels of Vitamin K-dependent Clotting Factors in the Chick

Two-week-old chicks adequate in vitamin K showed a relative lack of vitamin K-dependent clotting factors when compared with the rat, cow, and man. Chick prothrombin was 50%, IX 8%, and X 6% of respective levels in the rat. Factor VII was not detectable in chick plasma. When 1-day-old chicks were fed a vitamin K-deficient diet, prothrombin levels fell to 5% in 5 days, whereas factors IX and X fell to only 60% of normal. After warfarin administration to normal chicks, prothrombin levels fell to 20% in 6 hours, whereas factors IX and X fell to 60%. When cycloheximide was given to normal chicks, all vitamin K-dependent factors fell at the same relative rate with a half time of 2 hours. Cycloheximide also completely blocked the effect of physiological doses (10 mug) of phylloquinone upon prothrombin synthesis, but only partially blocked the effect of pharmacological doses (2.5 mg) of phylloquinone, suggesting an antagonism between cycloheximide and vitamin K at the ribosomal level. Puromycin was effective in blocking the action of vitamin K at both physiological and pharmacological doses. In the chick, unlike the rat, it appears that (1) cycloheximide is fully effective in blocking the action of physiological doses of vitamin K and (2) the regulatory systems for factors IX and X appear to have a higher affinity for vitamin K and a lower affinity for warfarin than the regulatory system for prothrombin.

Medium-Chain Triglycerides as Stress Agents in Vitamin K Deficiency

THE feeding of triglycerides containing fatty acids of medium chain length (MCT) creates a stress on vitamin E (Fisher and Kaunitz, 1965; Budowski et al., 1969). Continuing studies in this laboratory have indicated that the replacement of dietary long-chain triglycerides by MCT reduces the hypercholesterolemia of cholesterol-fed chickens (Fisher and Griminger, 1970). It was of interest to see whether one might use dietary MCT to obtain a more severe deficiency of another fat-soluble nutrient, vitamin K. To this end, the degree of vitamin K-deficiency in chickens receiving vitamin K-deficient diets containing MCT, corn oil, or no supplementary fats was compared, and the effect of supplementation of such diets with graded levels of different vitamin K sources investigated.PROCEDUREThe nitrogenous moiety in the vitamin K-deficient diet used in these tests was supplied (in percent) by isolated soy protein, 30, dl-methionine, 0.4, and glycine, 0.5. In addition, the diet contained reagent-grade…

Influence of sodium on calcium transport by the rat small intestine.

Influence of sodium on calcium transport by the rat small intestine.

Isolation of vitamin K from animal tissue

The isolation of phylloquinone from beef liver has been attempted. Development and use of a biological assay system has demonstrated that beef liver, beef liver lipid extracts, beef liver mitochondria, and the beef liver mitochondrial lipid extracts contain some compound that can alleviate elevated prothrombin times produced in chicks by a vitamin K-deficient diet. All attempts to isolate and identify this activity have been unsuccessful. The adoption of a modification of an isolation method developed by Fieser [ J. Am. Chem. Soc., 61, 3467 (1939)] facilitated the separation of two compounds from rabbit, chicken, and swine liver, one of which has been tentatively identified as a phylloquinone. The highest concentration was observed in rabbit liver (5 μg/gm lipid extract), and chicken liver contained a lower concentration (3.33 μg/gm lipid extract). Although absolute proof of the structure must await further work, the evidence thus far suggests that the compound may be 2-methyl-3-Δ 6-dehydrophytyl-1,4-naphthoquinone. The data obtained on the second compound isolated from rabbit, chicken, and swine liver do not permit any conclusion to be reached at present, but its spectral properties appear to be identical with those of α-tocopherolquinone.

Effect of Indigestible Oils on Vitamin K Deficiency in the Rat

Studies of indigestible oils such as mineral oil and squalene were undertaken to determine their effect on vitamin K deficiency in the rat. Severe hypoprothrombinemia occurred in male rats fed a vitamin K-deficient diet containing as little as 0.5% of the oil and the effect was reversed by feeding vitamin K. Female rats were partially resistant to the treatment. Dietary squalene also interrupted the assimilation of vitamin K in chicks where 0.5% of dietary oil inhibited the utilization of 0.1 µg of phylloquinone/g of diet. Similar results were obtained in chicks fed retinyl acetate and retinoic acid.

Comparative Vitamin K Activity of Dehydrated Alfalfa and Leucaena Leucocephala Meal

LEUCAENA LEUCOCEPHALA (Lam) de Wit, known in Hawaii as Koa Haole and previously identified by the generic name Leucaena glauca, is a legume widely distributed in tropical areas. Although the presence of the toxic amino acid, mimosine, may limit its potential use at high levels, satisfactory performance has been obtained when up to 5 percent was used in chick rations (Quisenberry and Draper, 1946; Molina, 1953). L. leucocephala is presently being used commercially in some of the Southeast Asian countries and it was in response to a request from this area that the present study was undertaken. MATERIALS AND METHODSOne hundred and sixty five one-day-old S.C. White Leghorn cockerel chicks were divided into eleven groups of 15 birds each and reared in an electric battery brooder with raised wire floors. During the first week all chicks received the following vitamin K-deficient diet: corn, 59.2%; soybean meal (44% protein), 36.0%;…

Relative Vitamin K Potency of Two Water-Soluble Menadione Analogues

MENADIONE sodium bisulfite, a water-soluble form of the provitamin 2-methyl-1,4-naphthoquinone, is used as a source of vitamin K in poultry feeds. This form differs in its vitamin K activity from the fat-soluble menadione (Frost and Spruth, 1955). The data presented in this report indicate that this difference is not necessarily a function, or, at least, not only a function of the change in solubility status, since another water-soluble form of menadione, namely, menadione dimethyl pyrimidinol bisulfite,2 differs in its activity from menadione sodium bisulfite. EXPERIMENTALFourteen groups of 20 cross-bred, female chicks received a purified vitamin K-deficient diet (Griminger, 1962a) from hatching to 1 week of age. At that time group uniformity was increased by eliminating 4 chicks with extreme weights. From 1 to 3 weeks of age the chicks received the K-deficient diet supplemented with 0.2% sulfaquinoxaline and with seven graded levels of each of the two water-soluble menadione…

Menadione Sodium Bisulfite Complex (Klotogen F® in Cecal Coccidiosis

ABSTRACT HEMORRHAGE is a primary, if not the principal, cause of death from cecal coccidiosis. Conversely, any reduction in the amount of hemorrhage might be expected to, at least, ameliorate the course of the disease. Harms and Tugwell (1956) reported that vitamin K activity, in the form of either Klotogen F or alfalfa meal, added to a basic vitamin K-deficient diet was capable of preventing deaths from cecal coccidiosis. Tugwell, Stephens and Harms 1957 provide further supporting data. The interpretation that the beneficial effects are due to the reduction in hemorrhage through the well known action of vitamin K is further supported by our unpublished evidence that neither menadione nor the water soluble menadione sodium bisulfite complex (Klotogen F) † have any coccidiocidal or coccidiostatic activity. The studies here reported were set up in the same basic pattern as the above, with only minor modifications. MATERIALS AND METHODS The basal diet was . . .