[Diethylamine malate in the treatment of asthma].
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Asthma↗
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[Effect of manganese (II), cobalt (II), and nickel (II) ions on the growth and production of coumarins in the suspension culture of Angelica archangelica L].
The plant cell reacts to an increased concentration of metals in the environment by various mechanisms. They include an increase in the formation of heat-shock proteins, metallothioneins, phytochelatins, amino acids (cysteine, histidine), organic acids (citric, malic), or secondary metabolites. The latter mechanism is being investigated for its possible use in explant cultures for the stimulation of secondary metabolism, which is the source of substances of pharmaceutical importance. The study tested manganese (II) (0, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, and 50 mM in the medium), cobalt (II), and nickel (II) ions (0, 0.1, 0.5, 1, 5, 10, 50, 100, 200, and 500 microM in the medium) as potential elicitors of coumarin production. At the same time, toxicity of these metals for the culture was examined by evaluating their effect on growth (characterized by fresh and dry weight of biomass at the end of a two-week cultivation). Cultures were cultivated in the dark and in the light. It has been found that the growth of cultures is not influenced by manganese in concentrations ranging from 0 to 2 mM, then it slightly decreases, at a concentration of 50 mM it is lower by 20 % when cultivated in the dark and by 30 % when cultivated in the light in comparison with the control. Cobalt in concentrations of 0 to 50 microM does not significantly influence the growth of the culture, higher concentrations decrease the biomass yields, more markedly when cultivated in the light (at 500 microM Co by 60 %, in the dark only by 30 % in comparison with the controls). Nickel in concentrations of 0.1 to 200 microM does not influence growth, and in a concentration of 500 microM decreases it by approximately 30 % in comparison with the control both in the light and dark. Production of coumarins was not stimulated by any metal in comparison with the control cultures, only the removal of manganese from the medium in the culture cultivated in the dark increased production by about 15 % versus the control.
Angelica archangelica↗
Antigenic analyses of the adult worms of Nippostrongylus brasiliensis by crossed immunoelectrophoresis.
In order to identify and to characterize the antigens from Nippostrongylus brasiliensis which are involved in the in vitro blastogenesis of T cells from infected animals, we have established by crossed immunoelectrophoresis a two dimensional antigenic map of the adult worm homogenate. Polyclonal precipitating rabbit antiserum recognized 26 antigens including five enzymes in these homogenates. The antigenic enzymes, the precipitates of which are only stained by specific staining, are: aldolase, malic enzyme, acid phosphatase, peroxydase and cholinesterase. This antigenic map enable us to identify 26 antigens in adult worm homogenates and to compare them with those which will be contained in purification fractions from these homogenates or in homogenates from other parasites.
Animals↗
Conformational stability of the N-terminal amino acid residues of mutated recombinant pigeon liver malic enzymes.
Pigeon liver malic enzyme has an N-terminal amino acid sequence of Met-Lys-Lys-Gly-Tyr-Glu-Val-Leu-Arg-. Our previous results indicated that the N-terminus of the enzyme is located at or near the enzyme's active center involved in Mn(II)-L-malate binding and is also near to the subunits' interface. In the present study, the conformational stability of the various deletion (delta) and substitution mutants at Lys2/Lys3 of the enzyme was investigated with chemical and thermal sensitivities. The lysine residue at position 2 or 3 seems to be crucial for the correct active site conformation, probably through an ion-pairing with Glu6. Deletion at Lys2 or Lys3, delta(K2/K3), and the double mutant K(2,3)E were much less stable than the wild-type enzyme towards chemical denaturation. Kinetic analysis of the thermal inactivation at 58 degrees C of the recombinant enzymes indicated that mutation at position 3 to alanine (K3A) endows the protein with extra stability compared with the wild-type enzyme. K3A is also stable towards chemical denaturation. The concentration of urea that causes half unfolding, [urea]0.5, for K3A is 3.25 M compared with 2.54 M for the wild-type enzyme. The K3A mutant of malic enzyme might therefore have potential practical applications.
Animals↗
Desaturation of polyunsaturated fatty acids in Mucor circinelloides and the involvement of a novel membrane-bound malic enzyme.
1. The component fatty acids of the endogenous phospholipids of microsomal preparations of Mucor, when shaken at 30 degrees C, increased in both chain length and in degree of unsaturation. The net effect was the production of gamma-linolenic acid which, over 2 h, increased from 17% to 32% of total fatty acids present. No further significant changes occurred after this time. 2. The major site for desaturation/elongation reactions was at the sn-2 position of PtdIns. PtdCho and PtdEtn were not implicated. 3. Of numerous metabolites and cofactors added to the microsomes, only malate could prolong the elongation/desaturation reactions for up to 6 h. This effect was shown to be due to a membrane-associated malic enzyme [malate dehydrogenase (decarboxylating) NADP+] with the NADPH produced being used in fatty-acid desaturation. 4. Kinetic analysis of cytosolic and microsomal enzymes [both in 0.1% (mass/vol.) Chaps] could not distinguish between them. However, when the microsomal malic enzyme was dialysed to remove Chaps, it lost 90% of activity, although the cytosolic malic enzyme lost only 20% activity. 5. The structural analogue of malate, tartronic acid, which is an inhibitor of malic enzyme, also inhibited the malate-induced stimulation of fatty-acyl group desaturation and elongation in the microsomal membranes. 6. It is concluded that two distinct malic enzymes exist, one soluble and one membrane bound, with similar active sites. Both have different roles in the production of NADPH, for lipid metabolism. The former will produce NADPH for fatty-acid biosynthesis whilst the latter produces NADPH for fatty-acid desaturation.
Cytochrome Reductases↗
Fatty acid profiles, lipogenesis, and lipolysis in lipid depots in finishing pigs treated with recombinant porcine somatotropin.
Thirty-two pigs (average weight, 74 kg) were used to determine the effects of recombinant porcine somatotropin (rpST) and dietary CP concentration on carcass and noncarcass tissues. Pigs were injected intramuscularly with either rpST (50 micrograms.kg BW-1.d-1; n = 16) or vehicle (n = 16) at 0900 for 24 d. Half the treated and control pigs were given ad libitum access to either a 14 or 20% CP corn-soybean meal diet. The left side of the carcass was physically dissected into separable skin, soft tissues, and bone. Tissue samples were obtained for enzyme assays, proximate analysis, and fatty acid profiles. Proximate analysis (fat, water, protein) of adipose tissue (AT) samples indicated that rpST decreased the percentage of ether extractable lipid (P < .01) and increased the percentage of protein and water (P < .05, P < .01) in intermuscular (IM), subcutaneous (SC), and intrafascicular (IF) AT depots and slightly altered fatty acid profiles of longissimus muscle IF and SC AT. Fatty acid synthesis and malic enzyme activity were decreased by rpST (P < .01), suggesting that lipogenesis was decreased; however, lipolysis was unaffected. Pigs fed the high-CP diet (20%) had decreased AT malic enzyme activity (P < .05) and fatty acid synthesis (P < .05). Additionally, pigs fed the high-CP diet had greater kidney weights (P < .01). Heart, liver, and kidney weights were heavier (P < .01) in pigs treated with rpST, whereas skin and total bone weight were unaffected. Neither weights nor lengths of four individual long bones were affected by dietary protein concentration or administration of rpST.
Adipose Tissue↗
Organic acids promote the uptake of lanthanum by barley roots.
Organic acids play an important role in metal uptake by, and accumulation in, plants. However, the relevant mechanisms remain obscure. Acetic, malic and citric acids increased the uptake of lanthanum (La) by barley (Hordeum vulgare) roots and enhanced La content in shoots under hydroponic conditions. Concentration-dependent net La influx in the absence and presence of organic acids yielded nonsaturating kinetic curves that could be resolved into linear and saturable components. The saturable component followed Michaelis-Menten kinetics. The K(m) values were similar; however, the V(max) values in the presence of acetic, malic and citric acids were 4.3, 2.8, 1.5-times that of the control, respectively. Enhanced uptake of La by organic acids was mediated mainly, but not solely, by Ca(2+) channels. X-ray absorption spectroscopic techniques provided evidence of La-oxygen environment and established that La(III) was coordinated to 11 oxygen atoms that are likely to be involved in the binding of La(III) to barley roots via carboxylate groups and hydration of La(III).
Acetic Acid↗
Comparative biochemical and immunological study of malic enzyme from two species of lactic acid bacteria: evolutionary implications.
Representatives of both Streptococcus faecalis and Lactobacillus casei produce isofunctional malic enzymes. All 10 strains of S. faecalis tested could be induced to synthesize malic enzyme and readily adapted to growth on malate. Although 17 of 21 L. casei strains could be induced to produce malic enzyme, only 9 of 14 strains tested grew at the expense of malate. A comparison of catalytic and regulatory properties suggested that the malic enzymes from S. faecalis and L. casei were very similar. Immunological analyses showed that the numerous similarities in function actually reflected partial protein homologies; however, two distinct forms of the malic enzyme were detected among different strains of L. casei by immunochemical and serological procedures. The division of L. casei into two subgroups based on the immunological type of malic enzyme synthesized corresponds to two subspecies currently recognized by microbial taxonomists.
Acrylates↗
Metabolism of sugars and organic acids in immature grape berries.
Individual intact excised immature Sultana berries were supplied through the cut pedicel with (14)C-sugars and organic acids. When (14)C-hexoses were supplied malic and tartaric acids accounted for 25% and 10% of the total activity extracted after 24 hours, and sucrose was synthesized. It is proposed that the changes in the levels of organic acids during ripening are related to changes in the ability of the berry to synthesize them. Although administration of uniformly labeled sucrose resulted in the unequal labeling of glucose and fructose, the results indicate breakdown of sucrose by invertase. It is suggested that the route of entry of the pedicel-fed sugars into the berry may be different from the route taken by sugar translocated from the leaf.
Journal Article↗
Hormonal regulation of the activity of the fatty acid synthesizing system and of the malic enzyme concentration in liver cells-1,2.
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Animals↗
Comparative effects of several chelating agents on the toxicity, distribution and excretion of aluminium.
The relative efficacy of citric, malic, malonic, oxalic and succinic acids, and deferoxamine mesylate (DFOA) on the toxicity, distribution and excretion in mice exposed to aluminum were compared. Chelating agents were administered intraperitoneally at a dose equal to one-fourth of their respective LD50. To determine the effect of the various chelators on the toxicity of aluminum, various doses of aluminum nitrate (938-3188 mg/kg) were administered intraperitoneally, followed by one of the chelators. Survival was recorded at the end of 14 days. Malic and succinic acids were the most effective. Malic acid and DFOA were the most effective in increasing the urinary excretion of aluminum. Citric acid was the most effective in increasing the faecal excretion of aluminum. Malonic, oxalic and succinic acids had no overall beneficial effects. Citric acid would appear to be the most effective agent of those tested in the prevention of acute aluminium intoxication. However, before the use of these compounds in human aluminium intoxication is possible, further investigations including the effects of these chelators after chronic aluminium intoxication are required.
Aluminum↗
The clinical and microbiological effects of a novel acidified sodium chlorite mouthrinse on oral bacterial mucosal infections.
Acidified sodium chlorite mouthrinses have been shown to have equivalent anti-plaque activity to those containing chlorhexidine, the current 'gold standard'. In this study, sodium chlorite mouthrinses (ASC) acidified with either malic or gluconic acids were compared to each other and with a chlorhexidine rinse and sterile water for their effect on salivary bacterial counts. Sixteen subjects participated in the study, which had a cross-over Latin square design. In a second study, a sodium chlorite/gluconic acid rinse was compared with chlorhexidine for its clinical and microbiological effects in 36 patients with oral mucosal infections. The sodium chlorite rinses acidified with malic and gluconic acids and the chlorhexidine rinse caused significant reductions in salivary bacterial counts up to 7 h after a single rinse compared with water. There were no significant differences between the three active treatments. In the mucosal infection study, there was a significant reduction in erythema in the chlorhexidine group compared with the ASC group. Patients who received the ASC rinse reported significantly less discomfort following treatment than those receiving the chlorhexidine rinse. Staphylococcus aureus counts were significantly reduced in the group who received the sodium chlorite rinse. There were no other significant differences between the treatments. ASC appears to be an effective alternative to chlorhexidine mouthrinse.
Adult↗
Metabolic flux analysis for succinic acid production by recombinant Escherichia coli with amplified malic enzyme activity.
A pfl ldhA double mutant Escherichia coli strain NZN111 was used to produce succinic acid by overexpressing the E. coli malic enzyme. Escherichia coli strain NZN111 harboring pTrcML produced 6 and 8 g/L of succinic acid from 20 g/L of glucose in flask culture at 37 degrees C and 30 degrees C, respectively. When NZN111(pTrcML) was cultured at 30 degrees C with intermittent glucose feeding the final succinic acid concentration obtained was 9.5 g/L and the ratio of succinic acid to acetic acid was 13:1. This system could not be analyzed by conventional metabolic flux analysis techniques, since some pyruvate and succinic acid were accumulated intracellularly. Therefore, a new flux analysis method was proposed by introducing intracellular pyruvate and succinic acid pools. By this new method the concentrations of intracellular metabolites were successfully predicted and the differences between the measured and calculated reaction rates could be considerably reduced.
Acetyltransferases↗
Photosynthetic Characteristics of Portulaca grandiflora, a Succulent C(4) Dicot : CELLULAR COMPARTMENTATION OF ENZYMES AND ACID METABOLISM.
The succulent, cylindrical leaves of the C(4) dicot Portulaca grandiflora possess three distinct green cell types: bundle sheath cells (BSC) in radial arrangement around the vascular bundles; mesophyll cells (MC) in an outer layer adjacent to the BSC; and water storage cells (WSC) in the leaf center. Unlike typical Kranz leaf anatomy, the MC do not surround the bundle sheath tissue but occur only in the area between the bundle sheath and the epidermis. Intercellular localization of photosynthetic enzymes was characterized using protoplasts isolated enzymatically from all three green cell types.Like other C(4) plants, P. grandiflora has ribulose 1,5-bisphosphate carboxylase and the decarboxylating enzyme, NADP(+)-malic enzyme, in the BSC. Unlike other C(4) plants, however, phosphoenolpyruvate carboxylase, pyruvate, Pi dikinase, and NADP(+)-malate dehydrogenase of the C(4) pathway were present in all three green cell types, indicating that all are capable of fixing CO(2) via phosphoenolpyruvate carboxylase and regenerating phosphoenolpyruvate. Other enzymes were about equally distributed between MC and BSC similar to other C(4) plants. The enzyme profile of the WSC was similar to that of the MC but with reduced activity in most enzymes, except mitochondrion-associated enzymes.Intracellular localization of enzymes was studied in organelles partitioned by differential centrifugation using mechanically ruptured mesophyll and bundle sheath protoplasts. Phosphoenolpyruvate carboxylase was a cytosolic enzyme in both cells; whereas, ribulose 1,5-bisphosphate carboxylase and NADP(+)-malic enzyme were exclusively compartmentalized in the bundle sheath chloroplasts. NADP(+)-malate dehydrogenase, pyruvate, Pi dikinase, aspartate aminotransferase, 3-phosphoglycerate kinase, and NADP(+)-triose-P dehydrogenase were predominantly localized in the chloroplasts while alanine aminotransferase and NAD(+)-malate dehydrogenase were mainly present in the cytosol of both cell types. Based on enzyme localization, a scheme of C(4) photosynthesis in P. grandiflora is proposed.Well-watered plants of P. grandiflora exhibit a diurnal fluctuation of total titratable acidity, with an amplitude of 61 and 54 microequivalent per gram fresh weight for the leaves and stems, respectively. These changes were in parallel with changes in malic acid concentration in these tissues. Under severe drought conditions, diurnal changes in both titratable acidity and malic acid concentration in both leaves and stems were much reduced. However, another C(4) dicot Amaranthus graecizans (nonsucculent) did not show any diurnal acid fluctuation under the same conditions. These results confirm the suggestion made by Koch and Kennedy (Plant Physiol. 65: 193-197, 1980) that succulent C(4) dicots can exhibit an acid metabolism similar to Crassulacean acid metabolism plants in certain environments.
Journal Article↗
Acute toxicity studies of aluminium compounds: antidotal efficacy of several chelating agents.
Four aluminum compounds--nitrate, chloride, sulphate and bromide--were administered orally and intraperitoneally to rats and mice. The LD50-values (14 days) were determined. The majority of deaths occurring during the first four days. The clinical and physical signs appearing after intoxication include among other lethargy, decreased locomotor activity, piloerection, weight loss and perorbital bleeding. After 14 days no alterations in liver and renal functions were detected in the animals which received intraperitoneally the LD50-values of aluminum nitrate as a single dose. Aluminum concentrations were highest in liver and spleen. No histopathological lesions could be observed. To compare the efficacies of nine chelating agents on the toxicity of aluminum in mice, the therapeutic index and the therapeutic effectiveness of each chelating agent have been calculated. Malic, succinic, oxalic and malonic acids showed the best results with malic and succinic acids being the most effective. Deferoxamine mesylate (DFOA), sodium salicylate, L-cysteine and citric acid were not so effective as antidotes for acute aluminum toxicity. Aurin tricarboxylic acid (ATCA) should not be used due to its high toxicity.
Alum Compounds↗
Production of succinic acid through overexpression of NAD(+)-dependent malic enzyme in an Escherichia coli mutant.
NAD(+)-dependent malic enzyme was cloned from the Escherichia coli genome by PCR based on the published partial sequence of the gene. The enzyme was overexpressed and purified to near homogeneity in two chromatographic steps and was analyzed kinetically in the forward and reverse directions. The Km values determined in the presence of saturating cofactor and manganese ion were 0.26 mM for malate (physiological direction) and 16 mM for pyruvate (reverse direction). When malic enzyme was induced under appropriate culture conditions in a strain of E. coli that was unable to ferment glucose and accumulated pyruvate, fermentative metabolism of glucose was restored. Succinic acid was the major fermentation product formed. When this fermentation was performed in the presence of hydrogen, the yield of succinic acid increased. The constructed pathway represents an alternative metabolic route for the fermentative production of dicarboxylic acids from renewable feedstocks.
Amino Acid Sequence↗
Potentiometric detection of organic acids in liquid chromatography using polymeric liquid membrane electrodes incorporating macrocyclic hexaamines.
Potentiometric detection employing coated-wire electrodes was applied to the determination of organic acids in liquid chromatography (LC). Poly(vinyl chloride)-based liquid membranes, incorporating lipophilic macrocyclic hexaamines as neutral ionophores were used as electrode coatings. The selectivity and sensitivity of the macrocycle-based electrodes were found to be superior to an electrode based on a lipophilic anion exchanger (a quaternary ammonium salt). Sensitive detection was obtained for the di- and tricarboxylic acids tartaric, malonic, malic, citric, fumaric, succinic, pyruvic, 2-oxoglutaric and maleic acids after separation in reversed-phase LC. Detection limits (signal/4sigmanoise=3) of 6 pmol for malonic acid and 2 pmol for maleic acid were attained. The detection was explained using a molecular recognition model. The hexaamine-based potentiometric electrodes had a 1-s response time at 1 ml min(-1) flow-rates. They were stable for at least 4 months, with an intra-electrode variation of 3.2% (n=5).
Acids↗
Enhancement effect study of some organic acids on the calcium availability of vegetables: application of the dynamic in vitro simulated gastrointestinal digestion method with continuous-flow dialysis.
The effect of added organic acids on the calcium availability of vegetables was investigated using the dialysis profiles obtained from an in vitro simulated gastrointestinal digestion with continuous-flow dialysis method. Citric acid was the most effective enhancer followed by tartaric, malic, and ascorbic acids. For amaranth, which has a low calcium availability (5.4%), a significant increase of availability was observed with increasing concentrations of all acids studied. With the continuous-flow dialysis approach, organic acids could be observed to promote the dialyzability even at an elevated intestinal pH. An enhancement effect from added organic acids was not clearly observed for Chinese kale, which itself contains a high amount of available calcium (52.9%).
Ascorbic Acid↗