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PubMed1988

New isozyme systems for maize (Zea mays L.): aconitate hydratase, adenylate kinase, NADH dehydrogenase, and shikimate dehydrogenase.

Electrophoretic variation and inheritance of four novel enzyme systems were studied in maize (Zea mays L.). A minimum of 10 genetic loci collectively encodes isozymes of aconitate hydratase (ACO; EC 4.2.1.3.), adenylate kinase (ADK; EC 2.7.4.3), NADH dehydrogenase (DIA; EC 1.6.99.-), and shikimate dehydrogenase (SAD; EC 1.1.1.25). At least four loci are responsible for the genetic control of ACO. Genetic data for two of the encoding loci, Aco1 and Aco4, demonstrated that at least two maize ACOs are active as monomers. Analysis of organellar preparations suggests that ACO1 and ACO4 are localized in the cytosolic and mitochondrial subcellular fractions, respectively. Maize ADK is encoded by a single nuclear locus, Adk1, governing monomeric enzymes that are located in the chloroplasts. Two cytosolic and two mitochondrial forms of DIA were electrophoretically resolved. Segregation analyses demonstrated that the two cytosolic isozymes are controlled by separate loci, Dia1 and Dia2, coding for products that are functional as monomers (DIA1) and dimers (DIA2). The major isozyme of SAD is apparently cytosolic, although an additional faintly staining plastid form may be present. Alleles at Sad1 are each associated with two bands that cosegregate in controlled crosses. Linkage analyses and crosses with B-A translocation stocks were effective in determining the map locations of six loci, including the previously described but unmapped locus Acp4. Several of these loci were localized to sparsely mapped regions of the genome. Dia2 and Acp4 were placed on the distal portion of the long arm of chromosome 1, 12.6 map units apart. Dia1 was localized to chromosome 2, 22.2 centimorgans (cM) from B1. Aco1 was mapped to chromosome 4, 6.2 cM from su1. Adk1 was placed on the poorly marked short arm of chromosome 6, 8.1 map units from rgd1. Less than 1% recombination was observed between Glu1 (on chromosome 10) and Sad1. In contrast to many other maize isozyme systems, there was little evidence of gene duplication or of parallel linkage relationships for these allozyme loci.

Aconitate Hydratase↗

PubMed1983

Fluorometric determination of carbodiimides with trans-aconitic acid.

A new fluorometric method for the determination of carbodiimides with trans-aconitic acid as a reagent has been developed. With the present method, as little as 50 pmol of carbodiimides can be determined. The relative standard deviations obtained with 25 nmol of dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide HCl (EDC) were 1.35, 1.41, and 2.72% (n = 10), respectively.

Acetates↗

PubMed1986

Trypanosoma brucei: cis-aconitate and temperature reduction as triggers of synchronous transformation of bloodstream to procyclic trypomastigotes in vitro.

Synchronous transformation of the monomorphic Trypanosoma brucei 427 variant clone MITat 1.4 (117) from bloodstream to procyclic trypomastigotes was studied in modified minimum essential medium plus 15% inactivated horse serum. Repression of variant surface glycoprotein synthesis, subsequent morphological transformation, and growth of procyclic cells was triggered by the simultaneous action of two signals: a reduction in temperature from 37 to 27 C and the addition of cis-aconitate. Repression of variant surface glycoprotein synthesis initiated by these two signals is reversible during the first hours, but becomes irreversible after about 1 day. Thereafter, cells are committed to differentiation at 27 C.

Aconitic Acid↗

PubMed2002

Induction of aconitate hydratase in hepatocytes of starving rats.

Induction of the activity of aconitate hydratase (AH) was observed in rat hepatocytes under the conditions of food deprivation. The increase in AH activity after 4 days of starvation in the studied tissues was from 0.57 to 2.05 U/g crude liver weight. The induction of aconitase was associated both with the cytoplasmic and mitochondrial AH isoforms. The activities of cytosolic and mitochondrial AH isoforms in starving animals consisted of 83 and 17% of the total activity, respectively. The cytoplasmic and mitochondrial isoforms of the enzyme with specific activities 11.1 and 6.13 U/mg protein, respectively, were obtained by a five-step purification procedure that included fractionation with ammonium sulfate, ion-exchanging chromatography on DEAE-Toyopearl and gel filtration. The purified preparations of these AH isoforms were electrophoretically homogenous. The molecular weights of these isoforms were estimated and several kinetic and regulatory properties were studied.

Aconitate Hydratase↗

PubMedDate not supplied

Mutagenic investigation of flavourings: dimethyl succinate, ethyl pyruvate and aconitic acid are negative in the Salmonella/mammalian-microsome test.

Three flavourings: dimethyl succinate, ethyl pyruvate and aconitic acid, commonly used in candy, beverages, and baked goods, were tested in the Salmonella/mammalian-microsome test. Tester strains were TA 1535, TA 100, TA 1537 and TA 98 and doses were 32, 160, 800, 4000 and 20 000 micrograms per plate. All tests were performed with and without the S9 fraction from Aroclor induced rat liver. None of the flavourings showed mutagenic potential. These results support the classification made by the Council of Europe, List I (1981).

Aconitic Acid↗

PubMed1994

Emendation of the description of Acidaminococcus fermentans, a trans-aconitate- and citrate-oxidizing bacterium.

Ruminal fluid which was enriched with trans-aconitate yielded a gram-negative diplococcus (strain AO) which was identified by 16S ribosomal DNA sequence analysis as Acidaminococcus fermentans. In contrast to the original description, the A. fermentans type strain and strain AO were found to utilize citrate as an energy source and to produce hydrogen and hydrogen sulfide. The descriptions of the genus and species are emended accordingly.

Aconitic Acid↗

PubMed1993

Experimental visceral leishmaniasis: role of trans-aconitic acid in combined chemotherapy.

We previously reported the effectiveness of trans-aconitic acid (TAA) as an antileishmanial compound. Inhibitory effects of TAA along with other antileishmanial compounds on transformation and in vitro multiplication in macrophage cultures of Leishmania donovani have been assessed. The efficacy of TAA in combined chemotherapy of experimental visceral leishmaniasis has also been evaluated along with those of commonly used antileishmanial compounds such as sodium stibogluconate, pentamidine, and allopurinol. TAA (2 mM) inhibited transformation of L. donovani amastigotes to promastigotes by 95.2%, whereas in combination with pentamidine (5 micrograms/ml), allopurinol (10 micrograms/ml), and sodium stibogluconate (50 micrograms of Sb per ml), it inhibited transformation by about 100, 99, and 98.5%, respectively. Sodium stibogluconate (20 micrograms of Sb per ml), pentamidine (2 micrograms/ml), and allopurinol (5 micrograms/ml) suppressed the amastigote burden in peritoneal macrophage cultures from BALB/c mice by 32.6, 56.1, and 46.3%, respectively. When these three drugs were used along with TAA (5 mM), the parasite loads were reduced by 100, 100, and 88.1%, respectively. TAA (5 mM) alone suppressed the amastigote burden by 59.5%. In experimental visceral leishmaniasis in hamsters (1-month model), TAA at a dose of 200 mg/kg of body weight per day suppressed the spleen parasite load by 73.5%, and TAA in combination with sodium stibogluconate (50 mg of Sb per kg per day), pentamidine (8 mg/kg/day), and allopurinol (15 mg/kg/day) inhibited the spleen parasite load by 98, 98.9, and 97%, respectively. Individually, these three drugs inhibited the parasite load by 35, 20, and 22%, respectively. TAA (400 mg/kg/day) inhibited the spleen parasite load by 99.8%, but an inhibitory effect of approximately 100% was noted when TAA was supplemented with an antileishmanial drug. TAA was administered in experimental animals through oral, intraperitoneal, and intramuscular routes; the intramuscular route was most effective.

Aconitic Acid↗

PubMed2006

The FAD-dependent tricarballylate dehydrogenase (TcuA) enzyme of Salmonella enterica converts tricarballylate into cis-aconitate.

Tricarballylate is the causative agent of grass tetany, a ruminant disease characterized by acute magnesium deficiency. Tricarballylate toxicity has been attributed to its ability to chelate magnesium and to inhibit aconitase, a Krebs cycle enzyme. Neither the ruminant nor the normal rumen flora can catabolize tricarballylate to ameliorate its toxic effects. However, the gram-negative enterobacterium Salmonella enterica can use tricarballylate as a carbon and energy source, providing an opportunity to study the genes and enzymes required for tricarballylate catabolism. The tricarballylate utilization (tcu) genes are organized into two transcriptional units, i.e., tcuR and tcuABC. Here, we report the initial biochemical analysis of TcuA. TcuA catalyzed the oxidation of tricarballylate to cis-aconitate. The apparent K(m) of TcuA for tricarballylate was 3.8 +/- 0.4 mM, with a V(max) of 7.9 +/- 0.3 mM min(-1), turnover number (k(cat)) of 6.7 x 10(-2) s(-1), and a catalytic efficiency (k(cat)/K(m)) of 17.8 M(-1) s(-1). Optimal activity was measured at pH 7.5 and 30 degrees C. The enzyme was inactivated at 45 degrees C. One mole of FAD was present per mole of TcuA. We propose a role for TcuB as an electron shuttle protein responsible for oxidizing FADH(2) back to FAD in TcuA.

Aconitic Acid↗

PubMedDate not supplied

[The use of liposomal form of phenylimide of cis-aconitic acid in herpes therapy].

The results of study of the antiviral activity and pharmacokinetics of phenylimide of cis-aconitic acid (PCAA) is presented. The 20% increase of the antiviral activity of PCAA incorporated into liposomes in comparison with the antiviral activity of the pure substance was shown. Liposomes with PCAA were tropic to lymphocytes and macrophages with maximum fluorescence being observed in the spleen, while empty liposomes were accumulated mainly in the liver. After the treatment with liposomal PCAA the symptoms of herpetic meningoencephalitis became less severe with 100% survival of the experimental animals. In the control group of rabbits 50% of the animals died, and in the surviving animals blindness or paralysis developed.

Aconitic Acid↗

PubMed2002

Electrospray ionization tandem mass spectrometric study of the aconitines in the roots of aconite.

Fragmentation pathways of aconitine-type alkaloids were investigated by electrospray ionization/ion trap multistage tandem mass spectrometry. Low-energy collision-induced dissociation of protonated aconitines follows a dominant first step, the elimination of the C(8)-substituent as acetic acid or fatty acid in MS(2) spectra. Successive losses of 1-4 CH(3)OH molecules, 1-3 H(2)O, CO, benzoic acid, and CH(3) or C(2)H(5) (N-substituents) are all fragmentation pathways observed in MS(3) and MS(4) spectra. By applying knowledge of these fragmentation pathways to the aconitines in the ethanolic extract of aconite roots, all the known aconitines were detected and also 23 unknown aconitine-type alkaloids, in which the lipo-alkaloids containing residues of 15C, 17C and 19C saturated or unsaturated fatty acids were characterized. These odd-carbon-number fatty acid substituents have not been reported previously.

Aconitine↗

PubMedDate not supplied

A case of fatal poisoning with the aconite plant: quantitative analysis in biological fluid.

In recent years recorded cases of plant poisoning have become rare, this may in part be due to the possibility of plant ingestion not being indicated at the beginning of an investigation. Aconitum napellus (aconite, Wolfsbane, Monkshood) is one of the most poisonous plants in the UK. It contains various potent alkaloids such as aconitine, isoaconitine, lycaconitine and napelline. Ingestion of Aconitum plant extracts can result in severe, potentially fatal toxic effects. This paper describes the analytical findings in a recent death in the UK. resulting from deliberate ingestion of Aconitum napellus extract. The concentrations of aconitine measured by HPLC-DAD in the post mortem femoral blood and urine were 10.8 micrograms/L and 264 micrograms/L, respectively. The aconitine concentration in the ante mortem urine was 334 micrograms/L and was estimated to be 6 micrograms/L in the ante mortem serum. Hence, accidental, suicidal or homicidal poisoning due to the ingestion of plant material remains a possibility and should be borne in mind when investigating sudden or unexplained death.

Aconitine↗

PubMedDate not supplied

Quantitative analysis of Aconitum alkaloids in the urine and serum of a male attempting suicide by oral intake of aconite extract.

A method for the quantitation of diesterditerpene-type Aconitum alkaloids and their hydrolysis products by gas chromatography-selected ion monitoring was applied to a clinical case study. A 45-year-old male attempted suicide by oral intake of Aconitum alkaloids, which are highly intoxicant extracts of Aconitum tubers. It was estimated that he had ingested approximately 11 mg of diesterditerpene-type alkaloids but was saved by intensive gastric irrigation. Mesaconitine, aconitine, hypaconitine, and their hydrolysis products were detected in the serum on the first day only. On the other hand, some alkaloids were still detectable in the urine even six days after intoxication. Aconitum alkaloids are biotransformed, and their hydrolysis products are excreted time-dependently to the urine. The urine was a useful material to identify the toxicants in the case of aconite intoxication.

Aconitum↗

PubMed1988

Effect of tricarballylic acid, a nonmetabolizable rumen fermentation product of trans-aconitic acid, on Mg, Ca and Zn utilization of rats.

trans-Aconitic acid has been implicated in magnesium deficiency of ruminants since the 1960s, but recent experiments indicated that much of it can be converted by rumen bacteria to tricarballylic acid (TCBA). Rats were used as experimental models in the studies described here because analogous experiments in ruminants would have been very expensive. When TCBA was fed to young male Sprague-Dawley rats as 2% of an AIN-76A diet with marginal (200 mg/kg) or adequate (500 mg/kg) Mg, virtually all of the dietary acid was recovered in the urine. Mg and Ca absorptions were unaffected by TCBA but urinary losses of Mg, Ca and Zn were higher than in pair-fed controls. TCBA was highly correlated (r = 0.93) with titratable acid excretion, and Mg, Ca and Zn retentions were reduced by 90, 35 and 56%, respectively. Rats fed 2% citric acid, a structurally similar but metabolizable acid, excreted more Zn than controls but did not excrete excess Mg or Ca. Since TCBA led to a depletion of Mg and other cations, it could be a contributing factor in the etiology of the Mg deficiency known as grass tetany in ruminants. Additional work in ruminants now seems warranted.

Administration, Oral↗

PubMed1981

Metabolism of trans-Aconitic Acid in Maize : I. PURIFICATION OF TWO MOLECULAR FORMS OF CITRATE DEHYDRASE.

Trans-aconitate synthesis via citrate dehydrase was determined in crude extracts of maize (Zea mays L.) coleoptiles. Two molecular forms of this enzyme were purified by substrate-specific elution from DEAE-cellulose, ammonium sulfate precipitation, and gel filtration. Each molecular form migrates as a single band in isoelectric focusing. Gel filtration and sodium dodecyl sulfate electrophoresis provided evidence that one enzyme form is composed of four 80,000-dalton subunits while the other is composed of two 60,000-dalton subunits. There was no evidence of proteolytic conversion of the large to the small molecular weight form when the former was incubated with either the 15,000(g) supernatant or with proteases. The data indicate that the two molecular forms of citrate dehydrase are isozymes.

Journal Article↗

PubMed1995

Depolarizing neuromuscular blocking action of coryneine derived from aconite root in isolated mouse phrenic nerve-diaphragm muscles.

The mode of the neuromuscular blocking action of coryneine (a quaternary ammonium derivative of dopamine) derived from aconite root was investigated in isolated phrenic nerve-diaphragm muscles and denervated diaphragm muscles of mice. Coryneine (20-150 microM) blocked the nerve-evoked twitch response without affecting the contraction evoked by electrical stimulation of the muscle. The blocking effect was reversed by neostigmine, a cholinesterase inhibitor. The electrical charge-response curve on depolarization produced by iontophoretically applied acetylcholine (ACh) at the endplate regions in normal muscles was shifted to the right on decreasing the maximal response by 40 microM coryneine. The double-reciprocal plot revealed that coryneine reduced the apparent affinity of ACh for its receptor on decreasing the maximal response. Coryneine (20 microM-2mM) itself depolarized the endplate membrane and this effect was reversibly suppressed by 1 and 5 microM pancuronium. Coryneine 30 microM-10mM) produced contractions of denervated muscles in a concentration-dependent manner and the effects were reduced by 70nM pancuronium. These results indicate that coryneine is a depolarizing agent and a mixed-type competitive and noncompetitive neuromuscular blocker.

Animals↗

PubMed1997

[The Aconit described by Nicander and today].

The herbal drug Aconite described by Nicander of Colophon (IInd century BC) is studied as to its symptoms and uses. The botanical identification presents some difficulties. There is a close correlation between the symptoms from Nicander's work and those cited by homeopaths authors.

Aconitum↗

PubMedDate not supplied

[Preclinical toxicologic study of a Baikal aconite tincture (bayacon)].

The safety of bayacon, a new drug based on Baikal aconite intended for the treatment of inflammation-proliferative dermatitis (psoriasis), was studied on a preclinical level. With respect to a single introduction in rats and mice, the drug is classified as a low-toxicity substance. However, a 3-month oral administration of bayacon (0.25, 0.5, and 2.5 mg/kg) in rats showed a number of dose-dependent functional and morphological changes. A dose of 2.5 mg/kg induced weak hyporegenerative anemia, neutrophile leukocytosis, and dystrophic changes in the stomach mucosa, heart, liver, and kidneys. All these symptoms disappeared within two weeks after abolition of the drug. Oral administration of bayacon (0.1 and 0.5 mg/kg) in rabbits produced no pathological morphofunctional changes in the organs and tissues studied. In rats, bayacon 2.5, 0.5, and 0.25 mg/kg doses of bayacon led to dose-dependent changes in some characteristics of the reproduction system. The drug did not influence the expression of allergic reactions and showed no immunotoxicity and mutagenicity manifestations.

Animals↗

PubMed2004

Effect of Shenfu Injection (ginesenoside and aconite alkaloid) on the apoptosis of intestinal mucosal epithelial cells and its mechanism during ischemia-reperfusion in rats.

OBJECTIVE: To investigate the effect of Shenfu Injection (SF, ginesenoside and aconite alkaloid) on the apoptosis of intestinal mucosal epithelial cells during ischemia-reperfusion in rats and its potential mechanisms. METHODS: Ischemia-reperfusion model was established in rats. Twenty-four rats were divided into 3 groups with 8 rats in each, eg, ischemia-reperfusion (I/R) group, SF-treated group, and control group. In both SF and I/R groups, the superior mesenteric artery was closed with forceps for 1 hour and then reperfused for 2 hours. Either SF (3 ml/kg, SF group) or normal saline (I/R and control groups) was injected intravenously and continuously for 5 ml/kg with a micropump before the superior mesenteric artery was closed. The superior mesenteric artery was not closed for animals in control group. The expression of casapse-3 and Fas, and the level of TNF-alpha and pathological changes of the ileal mucosal tissue were assayed. RESULTS: (1) The number of apoptosis cells increased obviously in I/R group and was significantly higher than that in SF and control groups (P<0.05). (2) The expression of caspase-3, Fas, and TNF-alpha was significantly higher in I/R group than SF and control groups (P<0.01); however, there was not significant difference in the expression of capase-3 between control group and SF group. There was a positive correlation between the expression of caspase-3, Fas, and TNF-alpha, and the number of apoptosis cells. (3) Under light microscope, intestinal mucosal impairment was found milder in SF group than I/R group (P<0.05). CONCLUSIONS: SF can depress the apoptosis of intestinal mucosal epithelial cells during ischemia-reperfusion by restraining the expression of TNF-alpha, Fas, caspase-3, and accordingly alleviate the ischemia and reperfusion injury of intestinal mucosal epithelial cells.

Animals↗