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The tissue distribution of L-ascorbic acid and dehydro-L-ascorbic acid in the guinea pigs injected intravenously with dehydro-L-ascorbic acid.

The tissue distribution of L-ascorbic acid (AsA) and dehydro-L-ascorbic acid (DAsA) in guinea pigs injected with DAsA intravenously was examined using high-performance liquid chromatography. DAsA injected into guinea pigs fed normal diets containing AsA (control group) was readily taken into erythrocytes, and AsA contents of plasma and other tissues rapidly increased after DAsA injection. In animals fed vitamin C-deficient diets, DAsA was also detected in erythrocytes; however, the increase of AsA in their tissues was considerably less than that of control group. From these results, it was suggested that utilization of DAsA as AsA in vitamin C-deficient guinea pigs was less than that of control animals, and the reduction mechanism of DAsA to AsA in vitamin C-deficient guinea pigs may have differed from that of control groups.

Administration, Oral↗

Ascorbic acid in thyroidectomized rats. II) Ascorbic acid status of the storage tissues and hepatic biosynthesis of glucuronic acid.

Depletion of ascorbic acid from adrenals, brain and epididymis along with loss in weight were noticed in the state of thyroidectomy. This decrease appears to be due to an effect of thyroidectomy on the membrane integrity since the membrane bound sialic acid was found to be significantly lowered in these tissues as a consequence of the elevated activity of sialidase. Thyroidectomy was also found to cause an adverse effect on the activities of hepatic UDP-glucuronyl transferase and beta-glucuronidase with no alteration in UDP-glucose dehydrogenase.

Adrenal Glands↗

[Acid-base equilibrium, blood lactic acid and pyruvic acid in albino rats after muscular exertion under conditions of normal oxygen, hypoxia and hypoxia-hypercapnia].

Albino rats of the Wistar family were subjected at three subsequent equal trials of muscular work: the first in normoxyc conditions; the second in hypoxic normobaric conditions; the third in hypoxic-hypercapnic normobaric conditions. The modifications of the lactacidemia, pyruvicemia and acid-base balance were greatest at the end of hypoxic trial. Added CO2 (2%) sensitively reduced the acidificant effects of the muscular work on the acid-base balance.

Acid-Base Equilibrium↗

[Effects of acute ingestion of ethanol on several metabolic parameters (blood sugar, triglycerides, non-esterified fatty acids, lactic acid and uric acid) in subjects with type I diabetes mellitus].

7 diabetic type I patients and 17 normal subjects underwent to acute ethanol oral load (0,8 g/Kg), in order to investigate the modifications produced on glucose, triglycerides, FFA, lactate and uric acid plasma values. Our results show basal differences between diabetics and controls in all these parameters; furthermore, acute ethanol ingestion produces no significant variations of blood glucose in both groups, a significant decrease in FFA levels, an increase of triglyceridemic levels, more evident in diabetics and an increase of lactate, earlier in normals. On the other hand, there is no significant modification in uric acid plasma levels in controls, whereas in diabetics there is an increase, probably due to more elevated lactacidemic values in the late phase of the test.

Adolescent↗

Can adults adequately convert alpha-linolenic acid (18:3n-3) to eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3)?

A diet including 2-3 portions of fatty fish per week, which corresponds to the intake of 1.25 g EPA (20:5n-3) + DHA (22:6n-3) per day, has been officially recommended on the basis of epidemiological findings showing a beneficial role of these n-3 long-chain PUFA in the prevention of cardiovascular and inflammatory diseases. The parent fatty acid ALA (18:3n-3), found in vegetable oils such as flaxseed or rapeseed oil, is used by the human organism partly as a source of energy, partly as a precursor of the metabolites, but the degree of conversion appears to be unreliable and restricted. More specifically, most studies in humans have shown that whereas a certain, though restricted, conversion of high doses of ALA to EPA occurs, conversion to DHA is severely restricted. The use of ALA labelled with radioisotopes suggested that with a background diet high in saturated fat conversion to long-chain metabolites is approximately 6% for EPA and 3.8% for DHA. With a diet rich in n-6 PUFA, conversion is reduced by 40 to 50%. It is thus reasonable to observe an n-6/n-3 PUFA ratio not exceeding 4-6. Restricted conversion to DHA may be critical since evidence has been increasing that this long-chain metabolite has an autonomous function, e.g. in the brain, retina and spermatozoa where it is the most prominent fatty acid. In neonates deficiency is associated with visual impairment, abnormalities in the electroretinogram and delayed cognitive development. In adults the potential role of DHA in neurological function still needs to be investigated in depth. Regarding cardiovascular risk factors DHA has been shown to reduce triglyceride concentrations. These findings indicate that future attention will have to focus on the adequate provision of DHA which can reliably be achieved only with the supply of the preformed long-chain metabolite.

Adult↗

Quantitative axial profiles of retinoic acid in the embryonic mouse spinal cord: 9-cis retinoic acid only detected after all-trans-retinoic acid levels are super-elevated experimentally.

Studies using bioassays in normal mice and gene activation in transgenic reporter mice have demonstrated peaks of retinoic acid receptor (RAR) signaling in the brachial and lumbar regions of the spinal cord. Recently, Solomin et al. (Solomin et al. [1998] Nature 395:398-402) detected a retinoid X receptor (RXR) signal in the same region of the developing spinal cord at a slightly later stage than the RAR signal. This finding raises the question of which retinoid ligands underlie RAR and RXR signaling in this part of the embryo. Quantitative measurements of regional differences in retinoid profiles have not been reported previously due to limitation in the sensitivity and specificity of available retinoid detection methods. Here, by using a recently developed ultrasensitive HPLC technique (Sakhi et al. [1998] J. Chromatogr. A 828:451-460), we address this question in an attempt to identify definitively the endogenous retinoids present in different regions of the spinal cord at the stages when regional differences in RAR and RXR signaling have been reported. We find a bimodal distribution of all-trans retinoic acid (at-RA), the ligand for RARs, and relate this to the expression of several retinoid-synthesizing enzymes. However, we do not detect 9-cis-retinoic acid (9-cis-RA), the putative RXR ligand, in any region of the spinal cord unless retinoid levels are massively increased experimentally by gavage feeding pregnant mice with teratogenic doses of at-RA. This study provides for the first time quantitative profiles of endogenous retinoids along the axis of the developing spinal cord, thereby establishing a foundation for more definitive studies of retinoid function in the future. It sets definite limits on how much 9-cis-RA potentially is present and demonstrates that at-RA predominates over 9-cis-RA by at least 30- to 180-fold in different spinal cord regions.

Aldehyde Oxidoreductases↗

Haematological effects in mice of the antitumour agents xanthenone-4-acetic acid, 5,6-dimethyl-xanthenone-4-acetic acid [correction of 5,6-methyl-] and flavone acetic acid.

Treatment of C57Bl/6 x DBA/2 mice with the maximal tolerated dose of flavone-8-acetic acid (FAA, 1300 mumol/kg), xanthenone-4-acetic acid (XAA, 1090 mumol/kg), or its dose-potent derivative 5,6-dimethyl-xanthenone-4-acetic acid (5,6-MeXAA, 100 mumol/kg) resulted within 24 h in a dramatic reduction in the number of circulating lymphocytes, an elevation in haemoglobin concentrations and a reduction in platelet numbers. Neutrophil counts either remained unchanged or were slightly elevated. All three compounds caused a marked loss of cells in the thymus. Examination of histological sections of thymus at 48 h following treatment with XAA revealed a selective depletion of cortical thymocytes and no effects on the epithelium or other thymic structures. A transient decrease in cell numbers was seen in the spleen and femoral bone marrow, with recovery to normal levels occurring within 3 days. The number of haemopoietic stem cells, colony-forming units in culture (CFU-c), in the femoral bone marrow increased after drug administration despite the occurrence of a decrease in the overall number of cells in the femur. In contrast to the increase in CFU-c numbers seen in vivo, 2 h exposure of bone-marrow cells to FAA, XAA or 5,6-MeXAA in vitro resulted in a decrease in the surviving fraction of CFU-c. The results are consistent with the hypothesis that the in vivo haematological effects of these compounds are indirect, perhaps being mediated through the induction of cytokines, and contrast with the haematological effects of conventional antitumour agents. The biochemical and haematological effects are unlikely to be the cause of the acute toxicity observed for these compounds.

Animals↗

Determination and metabolism of dithiol chelating agents. IV. Urinary excretion of meso-2,3-dimercaptosuccinic acid and mercaptosuccinic acid in rabbits given meso-2,3-dimercaptosuccinic acid.

The water-soluble dithiol chelating agents meso-2,3-dimercaptosuccinic acid (DMSA) and 2,3-dimercaptopropane-1-sulfonic acid (DMPS) are becoming of increasing importance for the treatment of lead, arsenic and mercury poisoning. There is, however, a paucity of data about their metabolic transformation. Male rabbits were given DMSA (0.20 mmol/kg) i.m., and urine was collected over a 6-hr period. Monobromobimane derivatization, HPLC separation, and fluorescence detection, along with [U-14C]DMSA data, demonstrated that the total 14C found in the urine was distributed as 73% unaltered DMSA, 7% mercaptosuccinic acid and 6 and 14% of two unknowns. Electrolytic reductive treatment of the urine did not increase the urinary content of DMSA, indicating that oxidative biotransformation is not a major pathway for DMSA in the rabbit. This latter result is strikingly different from that for DMPS in rabbit.

Animals↗

Analysis of methylphosphonic acid, ethyl methylphosphonic acid and isopropyl methylphosphonic acid at low microgram per liter levels in groundwater.

A method is described for determining methylphosphonic acid, ethyl methylphosphonic acid and isopropyl methylphosphonic acid, which are hydrolysis products of the nerve agents VX (S-2-diisopropylaminoethyl O-ethyl methylphosphonothiolate) and GB (sarin, isopropylmethyl phosphonofluoridate). The analytes are extracted from 50 ml groundwater using a solid-phase extraction column packed with 500 mg of silica with a bonded quaternary amine phase, and are eluted and derivatized with methanolic trimethylphenylammonium hydroxide. Separation and quantitation are achieved using a capillary column gas chromatograph equipped with a flame photometric detector operated in its phosphorus-selective mode. Two independent statistically-unbiased procedures were employed to determine the detection limits, which ranged between 3 and 9 micrograms/l, for the three analytes.

Chromatography, Gas↗

Enzymatic degradation processes of poly[(R)-3-hydroxybutyric acid] and poly[(R)-3-hydroxybutyric acid-co-(R)-3-hydroxyvaleric acid] single crystals revealed by atomic force microscopy: effects of molecular weight and second-monomer composition on erosion rates.

Enzymatic degradation processes of poly[(R)-3-hydroxybutyric acid] (P(3HB)) and poly[(R)-3-hydroxybutyric acid-co-(R)-3-hydroxyvaleric acid] (P(3HB-co-3HV)) single crystals in the presence of PHB depolymerase from Ralstonia pickettii T1 were studied by real-time and static atomic force microscopy (AFM) observations. Fibril-like crystals were generated along the long axis of single crystals during the enzymatic degradation, and then the dimensions of fibril-like crystals were analyzed quantitatively. The morphologies and sizes of fibril-like crystals were dependent on the molecular weight and copolymer composition of polymers. For all samples, the crystalline thickness gradually decreased toward a tip from the root of a fibril-like crystal after enzymatic degradation for 1 h. The thinning of fibril-like crystals may be attributed to the destruction of chain-packing structure toward crystallographic c axis by the adsorption of enzyme. From the real-time AFM images, it was found that at the initial stage of degradation the enzymatic erosion started from the disordered chain-packing region in single crystals to form the grooves along the a axis. The generated fibril-like crystals deformed at a constant rate along the a axis with a constant rate after the induction time. The erosion rate at the grooves along the a axis increased with a decrease of molecular weight and with an increase of copolymer composition. On the other hand, the erosion rate along the a axis, at the tip of the fibril-like crystal, was dependent on only the copolymer composition, and the value increased with an increase in the copolymer composition. The morphologies and sizes of fibril-like crystals were governed by both the erosion rates along the a axis at the grooves and tip of fibril-like crystals. In addition, we were able to estimated the overall enzymatic erosion rate of single crystals by PHB depolymerase from the volumetric analysis.

Crystallization↗

Dimers of formic acid, acetic acid, formamide and pyrrole-2-carboxylic acid: an ab initio study.

The intermolecular hydrogen bonds in dimers of formic acid, acetic acid, and formamide were investigated. Additionally, three configurations of the pyrrole-2-carboxylic acid (PCA) dimer were studied to analyze how the pyrrole pi-electron system influences the carboxylic groups connected by double O-H...O hydrogen bonds. The ab initio calculations for the systems investigated were performed at MP2/6-311++G(d,p), MP2/aug-cc-pVDZ, and MP2/aug-cc-pVTZ//MP2/aug-cc-pVDZ levels of theory. The "atoms in molecules" theory of Bader was used and the analysis of the critical points was performed to study the nature of hydrogen bonds. The decomposition of the total interaction energy applied here reveals that the delocalization energy term is a particularly important attractive contribution in these systems, more important in the case of systems forming homonuclear O-H...O double hydrogen bonds than in the case of those connected through heteronuclear N-H...O bonds. Because the systems analyzed may be formally classified as the resonance-assisted hydrogen bonds (RAHBs), it seems that the dominant contribution from the delocalization interaction energy term is a distinguished feature of such interactions.

Journal Article↗

Redox-active pH-responsive molecules: ferrocenylphosphonic acid, ferrocenylmethylphosphonic acid and 1,1'-ferrocenylbisphosphonic acid. Structural determination of FcPO3Na2.5H2O.

The acidity constants of the reduced and oxidized species of ferrocenylphosphonic acids FcPO3H2, FcCH2PO3H2 and fc(PO3H2)2 (Fc = (eta5-C5H5)Fe(eta5-C5H4), fc = (eta5-C5H4)Fe(eta5-C5H4)) in water have been evaluated by potentiometric, 31P NMR, and electrochemical methods. The oxidized forms are more acidic than the reduced ones. The interaction between the redox centre and the charged oxygen atoms of the phosphonate group is shown to be electrostatic. The maximum oxidation shift DeltaE between the protonated and unprotonated species increases with the number of charges of the substrate and decreases with the increase of the distance between the ferrocenyl centre and the oxygen atoms of the phosphonate group. The structure of FcPO3Na2.5H2O is determined. The compound crystallizes in the monoclinic system. It is lamellar with an inorganic layer formed by tetramers Na4O14, the ferrocenyl groups occupying the interlamellar space.

Journal Article↗

The acidic amino acids of tulip: isolation of gamma-ethylideneglutamic acid.

1. gamma-Ethylideneglutamic acid has been isolated from fruit capsules of tulip plants. 2. The assigned structure was indicated by examining the products formed after oxidation and catalytic hydrogenation and was confirmed by nuclear-magnetic-resonance spectroscopy and by synthesis of gamma-ethylglutamic acid. 3. The ability of gamma-ethylideneglutamic acid to participate in transamination and decarboxylation reactions was examined.

Chemical Phenomena↗

The structure and function of acid proteases. IV. Inactivation of the acid protease from Mucor pusillus by acid protease-specific inhibitors.

Mucor pusillus acid protease was rapidly inactivated with 1 : 1 stoichiometry by reaction with diazoacetyl-DL-norleucine methyl ester (DAN) in the presence of cupric ions. Cupric ions were essential for this inactivation. The rate of inactivation was maximal at around pH 6 when the enzyme was mixed with DAN and cupric ions without prior mixing of the reagents, and at pH 5.3 when DAN and cupric ions were mixed and incubated before addition to the enzyme solution. In both cases, the rate of inactivation decreased as the pH was either increased or decreased. The amino acid composition of an acid hydrolysate of the DAN-Modified enzyme was indistinguishable from that of the native enzyme except for the incorporation of about one norleucine residue per molecule of protein. The enzyme was also inactivated by reaction with 1,2-epoxy-3-(p-nitrophenoxy)-propane (EPNP). At the stage of about 90% inactivation, 1.50 residues of EPNP were incorporated per molecule of protein and the rate of inactivation followed pseudo-first order kinetics. The optimal pH for the inactivation was pH 3.0 and the rate of inactivation decreased as the pH was either increased or decreased. Furthermore, the enzyme was strongly inhibited by pepstatin, and the reactions of DAN and of EPNP was also inhibited significantly by prior treatment of the enzyme with pepstatin. These results suggest that the enzyme may have two essential carboxyl groups at the active site, one reactive with DAN in the presence of cupric ions and the other with EPNP, and that pepstatin binds part of the active site to inhibit the reactions with DAN and EPNP as well as the enzyme activity.

Aminocaproates↗

Mode of action of abscisic Acid in barley aleurone layers : abscisic Acid induces its own conversion to phaseic Acid.

As a part of our effort to study the mode of action of abscisic acid (ABA) and its metabolites during seed germination, we have investigated the regulation of ABA metabolism in barley (Hordeum vulgare) aleurone layers and a few other plant tissues. The rate of conversion of [(3)H]ABA to [(3)H]phaseic acid (PA), the first stable metabolite of ABA, is enhanced by 2- to 5-fold in barley aleurone layers when the tissue is pretreated with ABA. However, the conversion of [(3)H]PA to [(3)H] dihydrophaseic acid (DPA), the next metabolite after PA, is not enhanced by pretreatment with either ABA or PA. The ABA enhancement of its own metabolism in barley aleurone layers is detectable with a pretreatment of ABA ranging from 10(-3) to 10(-4) molar. This apparent self-induction of ABA conversion to PA can be observed after the barley aleurone layers have been treated with 10(-5) molar ABA for as short as 2 hours, and is inhibited by the transcription inhibitor, cordycepin (3'-deoxyadenosine), or the translation inhibitor, cycloheximide. The self-induction of ABA conversion to PA also occurs in wheat aleurone layers, but not in other plant tissues that have been investigated, including corn root tips, barley embryos, barley, and soybean leaf discs. It is probably a phenomenon unique to the aleurone layers of some cereal grains. In view of the recent observations that ABA is able to induce new proteins in barley aleurone layers, we suggest that some of these ABA-induced proteins are involved in the conversion from ABA to PA in this tissue.

Journal Article↗