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Intestinal absorption of alpha-tocopherol in the unanesthetized rat. The influence of luminal constituents on the absorptive process.

3H-alpha-tocopherol intestinal absorption was studied in the unanesthetized rat. The rate of alpha-tocopherol absorption remained linear over a wide range of concentrations (4 nM to 400 micrometer). Increasing the sodium taurocholate concentration in the micellar infusate up to 15 mM did not increase the rate of absorption of the vitamin. Addition of long-chain fatty acids to the micellar infusate decreased the absorption rate of the vitamin (p less than 0.05). The decrease was most significant (p less than 0.01) following the addition of the polyunsaturated linolenic (C18:3) acid. Increasing the hydrogen ion concentration in the perfusate increased the absorption rate of alpha-tocopherol. The present experiments in vivo support the conclusions drawn from in vitro uptake experiments which indicated that alpha-tocopherol is absorbed by a passive diffusion process. These experiments indicate that micellar expansion with polyunsaturated fatty acids interferes with the absorption of alpha-tocopherol and may result in deficiency of the vitamin.

Animals

Alpha-tocopherol: uses in preventing nitrosamine formation.

alpha-Tocopherol has been evaluated as a nitrite scavenger for the prevention of nitrosamine formation in a model system and under practical conditions. alpha-Tocopherol was found to react with nitrosating agents in both lipophilic and aqueous environments. The use of alpha-tocopherol was shown to inhibit aminopyrene-nitrite induced hepatotoxicity in rats and to reduce the amount of NDMA formed in cigarette smoke. Of primary interest is the finding of a significant reduction of NPYR formation in fried bacon. Nitrosamine inhibition was greater with alpha-tocopherol used in combination with sodium ascorbate than with sodium ascorbate alone.

Animals

[Vitamin E activity of some alpha-tocopherol derivatives and their effect on the ubiquinone level in rat liver in vitro].

An addition of alpha-tocopherol (I) and its synthetic derivatives (alpha-tocopheryl quinone (II), its short-chained analog (III), alpha-tocopherol lactone (IV), and short-chained alpha-tocopheryl acetate (V)) to the homogenized liver of vitamin E deficient rats resulted in a significant increase of ubiquinone after 2 hour incubation. Activity of the above derivatives (II-V) was not associated directly with their transformation into I or with a noticeable increase of the I content. There is a certain correlation between the chemical structure and the level of vitamin E activity of alpha-tocopherol derivatives that led to an increase in the ubiquinone content and prevented the decrease of tissue respiration and termination of pregnancy in rats.

Abortion, Veterinary

Articular cartilage preservation and storage. I. Application of tissue culture techniques to the storage of viable articular cartilage.

Articular cartilage slice explants were stored under various conditions, including freezing-thawing at various rates by using dimethyl sulfoxide (DMSO) as a cryoprotective agent, incubating in standard tissue culture medium (MEM Eagle:NCTC 135:15% fetal calf serum) in 5% CO2 and air at 4 degrees, 21 degrees, and 37 degrees C, and incubating in standard tissue culture medium containing 200 micrograms/ml alpha-tocopherol (vitamin E) at 37 degrees C after first ascertaining a dose-response curve of vitamin E. Results indicated that articular cartilage slice explants did not survive freezing or storage at 4 degrees and 21 degrees C as measured by 35S uptake. When stored at 37 degrees C in standard tissue culture in 5% CO2 and air, the slice explants remained viable for up to 60 days. The addition of alpha-tocopherol to the medium resulted in significantly less release of previously incorporated 35Sin stored cartilage slices and significantly less reduction of the amount of hexosamine present in the stored explants. alpha-Tocopherol in the medium also preserved safranin O staining. Thus, the application of tissue culture techniques to the storage of articular cartilage made it possible to preserve cartilage slice explants in a viable, biochemically "normal" state.

Animals

Vitamin E deficiency and seizures in animals and man.

Study of 100 children with grand mal convulsive disorders and 100 medically healthy children of matching age showed significantly lower plasma levels of vitamin E in the former (means 632.2 +/- 17.3 and 822.5 +/- 21.8 micrograms/dl respectively; p less than 0.001). This finding accords with the ability to prevent seizures in rodents by giving alpha-tocopherol before exposing them to a convulsion-inducing environment.

Animals

Contrasts in the actions of protein antibiotics on deoxyribonucleic acid structure and function.

The protein antibiotics neocarzinostain (NCS), macromomycin (MCR), and auromomycin (AUR), which is closely related to MCR, have been compared for their in vitro and in vivo actions on deoxyribonucleic acid (DNA). NCS, markedly stimulated by 2-mercaptoethanol, is much more active in inducing strand scissions in superhelical pMB9 and linear duplex lambda DNA than AUR, which is slightly inhibited by 2-mercaptoethanol. Purified MCR, even at very high levels, does not give any significant amount of cutting with either DNA substrate. 2-Propanol stimulates the activity of NCS but inhibits that of AUR. On the other hand, the antioxidant alpha-tocopherol strongly inhibits DNA breakage by both drugs. The intercalating drugs ethidium bromide, daunorubicin, proflavin, and actinomycin D at low concentrations inhibit DNA scission by AUR. The levels of intercalators required to inhibit NCS activity to comparable levels are about 10 times higher than those for AUR. Although MCR has virtually no in vitro DNA cutting activity, it is, like AUR and NCS, cytotoxic, as measured by the inhibition of DNA synthesis and induction of DNA strand breakage in HeLa cells.

1-Propanol

Activation and inactivation of neocarzinostatin-induced cleavage of DNA.

The possible role of free radicals in the mechanism of neocarzinostatin (NCS) action was studied. While mercaptene markedly stimulate the ability of NCS to degrade DNA, they also rapidly inactivate the antibiotic in a preincubation and at higher concentration inhibit the degradation reaction. The radiation protector S,2-aminoethylisothiuronium bromide-HBr is the most potent compound tested. Scavengers of diffusible OH radicals, O2- or H2O2 do not result in significant inhibition of the oxygen-dependent cleavage of DNA by NCS; in fact, alcohols and other organic solvents stimulate the reaction several-fold. By contrast, the potent peroxyl free radical scavenger, alpha-tocopherol, blocks the reaction 50% at 50 micron.

Alcohols

CRISPR-Cas9-mediated knockout of OsKCS11 in rice reveals potential crosstalk between very-long-chain fatty acids and cytokinin.

Very-long-chain fatty acids (VLCFAs) play crucial roles in various physiological processes in plants. Through our investigation using a CRISPR-Cas9 knockout mutant library in rice, we identified a semi-dwarf rice mutant named CRISPR-Cas-based dwarf-1 (csd-1). This mutant displayed multiple developmental defects, such as decreased plant height, panicle length, seed size, and seed-setting rate. Whole-genome resequencing analysis revealed that a T-nucleotide insertion in β-ketoacyl-CoA synthase 11 (KCS11), responsible for the initial step in fatty acid elongation, was responsible for the observed defects in csd-1. The identity of csd-1 was confirmed through genetic complementation and CRISPR-Cas9-mediated knockout. Expression analysis indicated that OsKCS11 was present in various tissues, with differential abundance observed through RT-qPCR and promoter GUS staining, and strong localization at the node position by RNA in situ hybridization; furthermore, OsKCS11 protein was confirmed to be in the endoplasmic reticulum. Furthermore, csd-1 exhibited significantly reduced levels of linolenic acid (18:3), C24:0-OH, C28:0-alkanes, C29:0-alkanes, alpha-tocopherol, and C33:0-alkanes, while trans-nonadecenoic acid and behenic acid levels were increased. Cytokinin analysis revealed significant increases in isopentenyladenine (IPA) and cis-zeatin (cZ) levels in csd-1. Molecular investigations indicated upregulation of genes involved in cytokinin biosynthesis or signaling, suggesting a potential link between VLCFAs and cytokinin synthesis through acetyl-CoA. This study not only proposed an alternative gene mapping method based on whole-genome resequencing but also elucidated the mechanism by which VLCFAs influence cytokinin synthesis and signaling.

Oryza

Effects of various compounds on lipid peroxidation mediated by detergent-solubilized rat liver NADPH-cytochrome C reductase.

A reconstituted lipid peroxidation system containing NADPH-cytochrome c reductase isolated from detergent-solubilized rat liver microsomes was used to determine the effects of several compounds, including drugs, on the lipid peroxidation activity. EDTA and ferrous ion were essential requirements for reconstitution of the activity. The addition of 1,10-phenanthroline to the system containing both EDTA and ferrous ion further enhanced the activity. Pyrocatecol, thymol, p-aminophenol, imipramine, p-chloromercuribenzoate (PCMB) and alpha-tocopherol exhibited strong inhibition, aniline, N-monomethylaniline, aminopyrine, benzphetamine, SKF 525-A and NADP exhibited moderate inhibition, and phenol, benzoic acid, acetanilide and nicotinamide exhibited less or no inhibition at the concentrations lower than 1000 micron M. Metal ions such as Hg+, Hg2+, Co2+, Cu2+, Mn2+ and U6+ inhibited lipid peroxidation strongly. In addition, Cd2+, St2+ and Ca2+ exhibited less potent to moderate inhibition, and Ba2+ and Mg2+ were without effects on the activity. Among sulfhydryl compounds tested, dithiothreitol inhibited lipid peroxidation to a greater extent than did the other three compounds, glutathione, cysteine and mercaptoethanol.

Amines

[Phytochrome system and synthesis of lipoquinones in the plastids of etiolated hordeum seedlings].

Short pulses of red light induce in etiolated barley seedlings an enhanced synthesis of plastidic benzoquinones and vitamin K1, which can be reverted by subsequent irradiation with short pulses of far-red. As compared to the dark there is more formation of plastoquinone-9 than alpha-tocopherol. The enhanced formation of vitamin K1 is coupled with a concomitant decrease in the level of the second naphthoquinone vitamin "K". The data show that active phytochrome Pfr, also in its ground state, induces the light triggered lipoquinone synthesis which is connected to thylakoid formation. The red light induction of enhanced plastoquinone-9 synthesis cannot be fully reverted by subsequent far-red and seems to be a very fast phytochrome response.

Chlorophyll

[The use of dried grape press cake in pig fattening].

In dried grape press cake the content of crude nutrients and ash, overall sugar, amino acids, alpha-tocopherol and gross energy was determined. In biological experiments with pigs (total of 109 animals) 10% of mixture A1 or SOL was replaced by the same amount of dried crushed grape press cake, without affecting negatively the weight gains and consumption of mixtures per unit of weight gain. Nutritional effects of grape press cake are a subject of discussion and comprise three factors: higher content of enrgy (fat and sugars) in mixtures containing press cake, anti-oxidation effect of press cake and the effect of tocopherols on the metabolism of basic nutrients.

Amino Acids

[Vitamins K and E and the functional state of connective tissue].

The function of the connective tissue in the skin of albino rats with the vitamin K deficiency was studied. Changes both in the cellular elements and in the fibrous structures (deranged differentiation of cellular elements, changes in the collagenformation, accumulation of acid mucopolysaccarides, etc.) were revealed. All of the mentioned changes are eliminated not only by introduction of vicasol, but also by that of alpha-tocopherol.

Animals

[Damage to the sarcoplasmic reticulum of skeletal muscles in leukemia: role of lipid peroxidation].

Ischemia development was accompanied by inhibition of the enzymatic transport system (ETS) of Ca2+ (reduction of the Ca2+/ATP value and of the Ca2+-dependent ATPase activity), this correlating with the accumulation of primary and secondary molecular products of lipid peroxidation (LPO) in the sarcoplasmic reticulum membranes of the skeletal muscles, in vivo. Administration of antioxidants (2,6-ditretbutyl-4-methylphenol, alpha-tocopherol) prevented the LPO activation in the ischemic muscle and partially protected the ETS of Ca2+ from damage. The blood supply restoration after prolonged ischemia led to further ETS of Ca2+ inhibition against the background of unchanges LPO products level.

Adenosine Triphosphatases

Antioxidant function of vitamin A.

Feeding of 100 000 I.U. of Vitamin A on alternate days, five times to rats, resulted in a marked lowering in in vitro lipid peroxidation of the tissue hemogenates. alpha-Tocopherol or DPPD supplementation along with A still further reduced the in vitro lipid peroxidation of the tissues. Vitamin A in large doses increased the antioxygenic potential of the tissues, and it is suggested that retinol also might be considered as a potential antioxidant similar to tocopherol in animal nutrition.

Alopecia

[Effect of vitamin D and vitamin E on the erythrocyte membrane. I. Effect on in vitro osmolar resistance and lysis].

As in the case of retinol, low doses of cholecalcipherol and alpha-tocopherol "in vitro" increase the osmotic resistance of red blood cells, while high doses have a lytic effect on the erythrocytes of various animal species (e.g.: man, sheep, rabbit, guinea-pig, mouse, rat) with the exception of the chicken. The haemolytic effect of cholecalcipherol is less intense than that of retinol; alpha-tochopherol, while causing even less haemolysis than cholecalcipherol, involves a remarkable loss of K+ from the red cells. Besides these quantitative differences, the mechanism of action of the three vitamins seems somewhat different, as shown by modifications of the suspension medium (e.g.: pH, temperature, presence of proteins and reducing substances) or by treatment of the red cells with trypsin. Our results therefore essentially confirm the hypothesis of Dingle and Lucy of common mechanism of action of liposoluble vitamins on biological membranes.

Cholecalciferol

Comparison of hepatic microsomal drug-metabolizing systems from rats fed crude and purified diets.

Hepatic microsomes from rats fed a crude or a purified diet were compared by measureing their contents of protein, cytochrome P-450, and cytochrome b5, their rates of activity of NADPH- and NADH-cytochrome c reductases, NADPH-cytochrome P-450 reductase, NADPH oxidase, lipid peroxidase, ethylmorphine N-demethylase, aniline hydroxylase, benzpyrene hydroxylase, and their substrate-binding spectra (ethylmorphine, hexobarbital, aniline, and ethyl isoyanide). With the exception of lipid peroxidase activity, which was much higher in microsomes from animals fed the crude diet, little or no consistent diet-related differences in these measurements were observed over a 4-week experimental period, nor were results significantly less variable with one or the other diet. No consistent significant differences were observed with two strains of rats. The lower lipid peroxidase activity seen with the purified diet appeared to be due to the high vitamin E intake when that diet was employed; rats fed the crude diet and an oral supplement of alpha-tocopherol yielded microsomes with low lipid peroxidase activities similar to those seen in microsomes from rats fed the purified diet. A gradual temporal increase in benzpyrene hydroxylase activity was observed with both diets. This was interpreted to be due to environment inducing agents other than those present in the diet.

Analysis of Variance