Effect of threonine and glycine concentrations on threonine aldolase activity of yogurt microorganisms during growth in a modified milk prepared by ultrafiltration.

To evaluate the combined effects of threonine and glycine concentrations during growth on threonine aldolase activity (EC 2.1.2.1) of yogurt microorganisms, Streptococcus thermophilus and Lactobacillus bulgaricus, a modified milk growth medium was prepared using UF to deplete the free amino acid level. Threonine and glycine were added according to a 2x2x2 factorial design at 5 or 195 microg.ml(-1) along with a standard amino acid mixture. Acetaldehyde production and threonine aldolase activity were evaluated utilizing headspace gas chromatography. Results showed that threonine and glycine concentrations did not affect growth or titratable acidity. The high concentration of threonine in combination with low glycine in the growth medium resulted in increased acetaldehyde synthesis by both microorganisms. Conversely, high glycine with low threonine decreased acetaldehyde synthesis. High threonine and low glycine increased threonine aldolase activity of cell-free extracts from S. thermophilus and L. bulgaricus, whereas high glycine and low threonine reduced threonine aldolase activity of both microorganisms.

Blockade of ethanol induced conditioned taste aversion by 3-amino-1,2,4-triazole: evidence for catalase mediated synthesis of acetaldehyde in rat brain.

This investigation seeks to present evidence for the oxidation of ethanol in the brain via the peroxidatic activity of catalase and simultaneously provide evidence for the role of central acetaldehyde (ACH) in the mediation of an ethanol-induced conditioned taste aversion (CTA). Ethanol is capable of inducing a conditioned taste aversion. Pretreatment with the catalase inhibitor, 3-amino-1,2,4-triazole (AT), shows an attenuation of this ethanol-induced CTA. Animals receiving ethanol injections showed a CTA to a novel solution paired with a drug administration, while ethanol injected animals pretreated with AT did not show a CTA to ethanol administration. This effect of AT appears to be specific to the effects of ethanol as CTA's to morphine and lithium chloride were not affected by AT pretreatment. Peripheral levels of ethanol were the same in all animals regardless of pretreatment indicating that AT had no effect on peripheral levels of ethanol. These data increase support for the notion that acetaldehyde is produced directly in the brain and that it may be the agent mediating some of the psychopharmacological properties of ethanol.

Metabolism and relative carcinogenic potency of chloroethanes: a quantum chemical structure-activity study.

Using the all-valence electron, semiempirical molecular orbital method, MNDO, properties have been identified and calculated for eight chloroethanes which can serve as indicators of their extent of transformation to alcohols by cytochrome P450 and the subsequent formation of aldehydes by loss of HCl from these alcohols. The assumption was made that these aldehydes are the active carcinogens of the chloroethanes and that they act as electrophiles in adduct formation with DNA bases. Electrophilic properties of these putative ultimate carcinogens have been calculated which are indicators of the rank order of carcinogenic activity of the parent compounds in susceptible species. Particularly relevant in this respect are (a) the electron affinity of aldehydes as measured by the energy of their electron accepting (lowest energy virtual) orbital, and (b) the net charge on the C alpha carbon, adjacent to the carbonyl carbon, which can participate in electrophilic attack on nucleophilic sites of DNA bases. The molecular properties identified in this study as indicators of rank order or carcinogenic activity of the parent chloroethanes are consistent with the importance of cytochrome P450 in transforming halohydrocarbons to active carcinogens and of acylchlorides and chloroaldehydes as the active form. Their validity and usefulness can be further tested in screening unknown and more complex chlorohydrocarbons for carcinogenic activity.

Possibility of diacetyl and related compounds as the 4-carbon compound necessary for the formation of riboflavin in Ashbya gossypii.

The effects of various compounds (0.5%) involved in the butanediol and the glycolytic pathways on riboflavin formation in whole cells of Ashbya gossypii at rest were examined. The addition of acetate, glycerol and diacetyl inhibited riboflavin formation, while the addition of acetoin had no effect on it, and the addition of ethanol, 2,3-butanediol, pyruvic acid and glucose accelerated it. The relation of diacetyl and acetoin to riboflavin formation during resting cell incubation in the presence of 0.5% ethanol and various concentrations of 2,3-butanediol was examined. The results quantitatively revealed a precursor-product relation between riboflavin formation and the formation of diacetyl and acetoin. The results obtained provide evidence that a high flavinogenic agent, ethanol, was converted to acetaldehyde, pyruvic acid, acetoin and diacetyl in this order, that a week flavinogenic agent, 2,3-butanediol, was transferred to diacetyl through acetoin, and that the diacetyl produced can be utilized as the 4-carbon compound for riboflavin formation in the flavinogenic mold, Ashbya gossypii. It remains obscure whether diacetyl is enzymatically involved in riboflavin formation.

Metabolism of paraldehyde to acetaldehyde in liver microsomes. Evidence for the involvement of cytochrome P-450.

A concentration-dependent acetaldehyde (AcH) generation was observed when paraldehyde was incubated with the mouse liver microsomal fraction. The process, which exhibited a requirement for oxygen and NADPH and was inhibited by carbon monoxide, was found to have a Km of 17.9 mM with respect to paraldehyde and a Vmax of 40.1 nmoles/mg protein/min with respect to AcH formation. NADH was much less effective as an electron donor than NADPH, though a more than additive increase in AcH generation was observed when both of these nucleotides were added to the incubation. The rate of microsomal AcH generation from paraldehyde was increased 2.5-fold by pretreatment of the mice with phenobarbital but only 0.6-fold by pretreatment with 3-methylcholanthrene. Pretreatment with 2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride (SKF-525A) resulted in 54% inhibition of the reaction rate. Addition of metopirone to the incubation inhibited AcH generation in a concentration-related fashion, the inhibition being greatest, proportionately, in microsomes from phenobarbital-pretreated animals. The above results conclusively indicate the involvement of cytochrome P-540 mixed function oxidase in the formation of AcH from paraldehyde by mouse liver microsomes. It is also postulated that this process may be accomplished in the reaction analogous to O-dealkylation.

Binding of metabolically derived acetaldehyde to hepatic proteins in vitro.

Homogenates of rat liver incubated with 10 mM [14C]ethanol were analyzed for acetaldehyde production and for both stable and unstable radiolabeled acetaldehyde adducts with proteins. During incubation, formation of acetaldehyde and of 14C-labeled proteins both increased with time in a parallel manner. Acetaldehyde generation and subsequent formation of radiolabeled proteins were potentiated by supplementation of the cell-free system with NAD+ (1 mM). Cycloheximide (0.1 mM) caused no significant reduction in protein-bound radioactivity, whereas the addition of strong nucleophiles L-cysteine (5 mM) and reduced glutathione (5 mM) each decreased radiolabeling by 50 to 60%. Preheating of crude homogenates at 90 degrees C, prior to incubation with [14C]ethanol profoundly decreased subsequent production of acetaldehyde and formation of 14C-labeled proteins. The results indicate that the major source of protein-bound radioactivity derived from [14C]ethanol oxidation in this system is due to binding of enzymatically derived [14C]acetaldehyde to hepatic proteins.

The pathogenesis of hepatitis in alcohol abuse and jejunoileal bypass.

Acetaldehyde, produced in excessive amounts in alcohol abuse or after jejunoileal bypass, binds to hepatocyte plasma membranes by way of formation of an intermediate Schiff's base. This binding has no effect on cellular metabolism or membrane function but causes physical changes in membrane properties which activate the complement cascade and this results in hepatocellular damage.

Blood acetaldehyde levels in alcohol-dosed rats after treatment with ANIT, ANTU, dithiocarbamate derivatives, or cyanamide.

In adult female Wistar rats, pretreated by gavage with two doses - 16 or 256 mumol/kg - of cyanamide, TMTD (tetramethylthiuram disulfide), TMTM (tetramethylthiuram monosulfide), Ziram or Zineb at 90 min or 18 h before administration of 2 g of ethanol/kg i.p., the blood acetaldehyde levels were significantly increased for 90 - 240 min after ethanol administration (exceptions were noted after exposure to Zineb for 90 min or to low-dosed cyanamide for 18 h). After pretreatment for identical periods with ANTU (N-1-naphthylthiourea) or ANIT (1-naphthylisothiocyanate) at doses extending into the LD50 range, the blood acetaldehyde levels of rats given the same dose of ethanol remained uninfluenced. The increase in blood acetaldehyde recorded after 16 mumol/kg p.o. of TMTM and TMTD remained detectable for up to 48 h. Onset of the cyanamide action occurred already after 45 min. While recognizing that results from animal experiments cannot be transposed without restriction to the human situation, it is concluded that occupational contacts with ANTU or ANIT are not likely to elicit increased blood acetaldehyde levels in man after ingestion of alcohol. The risk of an ethanol intolerance reaction due to a rise in blood acetaldehyde therefore does not appear to be warranted. The present findings indicate, however, that exposure to TMTD, TMTM, Ziram, Zineb or cyanamide is associated with a definite health risk; because of the long persistence of these substances in the body, the risk exists for a long time post-exposure.

[Biological profile of liver damage in alcoholics (author's transl)]. / Profil biologique de l'atteinte hépatique chez l'éthylique.

It is now possible to pinpoint the biochemical processes responsible for liver damage in alcoholics and to monitor detoxication from a biological point of view. Cirrhosis of the liver is a direct consequence of chronic alcoholism in 60-80 % of cases, and most alcoholics, after several years of drinking, cannot escape the ravages of alcohol, although it is not yet known why a small proportion are not affected biochemically. Psychological deterioration can be explained biochemically, due to the neurotoxic effects of acetaldehyde, formed by alcohol but 20-30 times more toxic, which acts on catecholamines and serotonin with, inter alia, depressant, habit-forming, hallucinogenic and convulsive properties. Hepatic symptoms depend on the stage the alcholic has reached - acute, subacute or the chronic and final stage of cirrhosis of the liver, and include disturbances of transaminases, gamma-GT, serum proteins, immunoglobulins, specific proteins, lipids (including very interestingly an increase in cholesterol-HDL which needs to be investigated further) and hematic changes with FDP and thrombocytes affected, and often anaemia.

[Chemical mechanism of the effect of alcohol]. / Chemische Mechanismen der Alkohol-Wirkung.

The formation of tetrahydro-isoquinoline alkaloids in human and animal metabolisms subsequent to alcohol consumption is reported. The spectrum of identified products ranges from simple structures such as salsolinol to the complicated skeleton of protoberberines. These substances, hitherto found exclusively in plants, interfere with the biosynthesis, biodegradation and neurotransmission of the structurally closely related biogenic amines from which they are formed, and thus affect the vegetative nervous system.

Microbial metabolism of phenolic amines: degradation of dl-synephrine by an unidentified arthrobacter.

Microorganisms capable of degrading dl-synephrine were isolated from soil of Citrus gardens by enrichment culture, with dl-synephrine as the sole source of carbon and nitrogen. An organism which appears to be an arthrobacter, but which cannot be identified with any of the presently recognized species was predominant in these isolates. It was found to metabolize synephrine by a pathway involving p-hydroxyphenylacetaldehyde, p-hydroxyphenylacetic acid, and 3,4-dihydroxyphenylacetic acid as intermediates. Some of the enzymes of this pathway were demonstrated in cell-free extracts. An aromatic oxygenase, which could also be readily obtained in a cell-free system, was found to degrade 3,4-dihydroxyphenylacetic acid by meta cleavage.

Novel pathway for degradation of protocatechuic acid in Bacillus species.

A species of Bacillus, tentatively identified as B. circulans, degrades protocatechuic acid by a novel reaction involving meta-fission between C2 and C3 of the benzene nucleus. 2-Hydroxymuconic semialdehyde is then degraded to pyruvate and acetaldehyde by enzymatic reactions described in previous work. Protocatechuate 2,3-oxygenase exhibits a rather narrow substrate specificity; the methyl and ethyl esters of protocatechuic acid are oxidized, but other substrates for ring-fission oxygenases, notably catechol, gallic acid, and homoprotocatechuic acid, are not attached.