Ethanol in formularies of US teaching hospitals.
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Alcohol Withdrawal Delirium↗
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Changes in myocardial metabolism induced by ethanol administration.
The utilization of ethyl alcohol in the myocardium and the reactions connected with its metabolism were investigated in rabbits which were given over a period of two months a 30% ethanol solution per os in a dosage of 1 g/kg body weight, and in a control group. Alcohol intoxication led to a reduction of the reserve of oxidized nicotinamide coenzymes, to an intensification of glycolytic processes, to slowed down utilization of substrates in the citric acid cycle and to an accumulation of less-oxidized metabolism products (dihydroxyacetone phosphate, lactate, pyruvate, malate, oxaloacetate, 2-oxoglutarate). A change was also observed in the myocardial microstructure. Metabolic changes are evidently one of the links of the pathogenetic mechanism of alcohol intoxication, which cause disorders in cardiac activity diagnosed as alcoholic myocardial dystrophy.
Alcoholism↗
Experimental investigations of multiple steady states in aerobic continuous cultivations of Saccharomyces cerevisiae.
The steady-state behavior of a glucose-limited, aerobic, continuous cultivation of Saccharomyces cerevisiae CEN.PK113-7D was investigated around the critical dilution rate. Oxido-reductive steady states were obtained at dilution rates up to 0.09 h(-1) lower than the critical dilution rate by operating the bioreactor as a productostat, where the dilution rate was controlled on the basis of an ethanol measurement. Thus, the experimental investigations revealed that multiple steady states exist in a region of dilution rates below the critical dilution rate. The existence of multiple steady states was attributed to two distinct physiological effects occurring when growth changed from oxidative to oxido-reductive: (i) a decrease in the efficiency of ATP production and utilization (at ethanol concentrations below 3 g/L) and (ii) repression of the oxidative metabolism (at higher ethanol concentrations). The first effect was best observed at low ethanol concentrations, where multiple steady states were observed even when no repression of the oxidative metabolism was evident, i.e., the oxidative capacity was constant. However, at higher ethanol concentrations repression of the oxidative metabolism was observed (the oxidative capacity decreased), and this resulted in a broader range of dilution rates where multiple steady states could be found.
Adaptation, Physiological↗
Studies of diacylglycerol cholinephosphotransferase and diacylglycerol ethanolaminephosphotransferase activities in Tetrahymena microsomes.
Microsomes isolated from Tetrahymena pyriformis synthesized phosphatidylcholine and phosphatidylethanolamine by CDPcholine: 1,2-diacylglycerol cholinephosphotransferase (EC 2.7.8.2) and CDPethanolamine: 1,2-diacylglycerol ethanolaminephosphotransferase (EC 2.7.8.1), utilizing ethanol-dispersed dioleoglycerol. Cholinephosphotransferase and ethanolaminephosphotransferase activities have similar dependences on MgCl2 and MnCl2, but the latter was more effective than the former for both enzyme activities. The V values for 1,2-dioleoylglycerol obtained at optimal conditions were 1.8 nmol/min per mg microsomal protein for cholinephosphotransferase and 0.6 nmol/min per mg microsomal protein for ethanolaminephosphotransferase. Both enzymes could not utilize 1,3-dioleoylglycerol or 1-oleoylglycerol as substrates. Cholinephosphotransferase had an apparent Km for CDPcholine of 11.7 microM with 1,2-dioleoylglycerol and was inhibited by CDPethanolamine competitively. On the other hand, ethanolaminephosphotransferase has an apparent Km for CDPethanolamine of 8 microM and CDPcholine was a noncompetitive inhibitor of ethanolaminephosphotransferase activity. Furthermore, despite the marked alteration of phospholipid composition occurring during the temperature acclimation of Tetrahymena cells, both enzyme activities showed similar dependences on growth and incubation temperatures. This may imply that the final step of de novo synthesis of two major phospholipids does not participate in the thermally induced modification of the profile of phospholipid polar head group in membranes.
Animals↗
[Identification of partially deuterated exometabolites during culture of tea fungi in heavy water].
Exometabolites of tea fungus during the cultivation on heavy water were studied by methods of high-resolution proton and deuterium exchange. Is was shown that, depending on the degree of exchange of proton for deuterium in the methyl group of acetate, three isotopomers exist: -CH3, -CH2D, and -CHD2. Ethanol is represented by six isotopomers: -CHD-CH3, -CD2-CH3, -CD2-CH2D, -CD2-CHD2, -CHD-CH2D, and -CHD-CHD2. Relative concentration of these isotopomers and the percent content of deuterium were determined. Based on these data, the kinetics of metabolism of the object cultivated on heavy water was analyzed. It was show that the efficiency of glucose utilization is the same as during cultivation on light water with the exception that the time of utilization is significantly greater. At the same time, the parameters of utilization of ethanol and its conversion into acetic acid in the life activity of the tea fungus in heavy and light water are very similar.
Deuterium↗
Significant pathways of hepatic ethanol metabolism.
Rat liver microsomes oxidized ethanol two to three times faster than propanol when incubated with either an NADPH- or an H2O2-generating system. In addition, solubilized, purified microsomal subfractions were found to contain protein with an electrophoretic mobility identical to rat liver catalase on SDS polyacrylamide gels, suggesting that the separation of catalase from cytochrome P-450 and other microsomal components may not be feasible. These data support the postulate that catalase is responsible for NADPH-dependent microsomal ethanol oxidation. Direct read-out techniques for pyridine nucleotides, the catalase-H2O2 complex, and cytochrome P-450 were utilized to evaluate the specificity of inhibitors of alcohol dehydrogenase (4-methylpyrazole; 4 mM) and catalase (aminotriazole; 1.0 g/kg) qualitatively in perfused rat livers. 4-Methylpyrazole and aminotriazole are specific inhibitors for alcohol dehydrogenase and catalase, respectively, under these conditions. Neither inhibitor nor a combination of them altered the mixed function oxygen of p-nitroanisole to p-nitrophenol as observed by oxygen uptake and product formation. When ethanol utilization was measured over the concentration range 20-80 mM in perfused liver, a concentration dependence was observed. At low concentrations of ethanol, ethanol oxidation was almost totally abolished by 4-methylpyrazole; however, the contribution of 4-methylpyrazole-insensitive ethanol uptake increased as a function of ethanol concentration. At 80 mM ethanol, ethanol utilization was nearly 50% methylpyrazole-insensitive. This portion of ethanol oxidation, however, was abolished by aminotriazole. The data indicate that alcohol dehydrogenase and catalase-H2O2 are responsible for hepatic ethanol oxidation. At low ethanol concentrations (less than 20 mM), alcohol dehydrogenase is predominant; however, at higher ethanol concentrations (up to 80 mM), the contribution of catalase-H2O2 to overall ethanol utilization is significant. No evidence that the endoplasmic reticulum is involved in ethanol metabolism in the perfused liver emerged from these studies.
1-Propanol↗
The yeast CPC2/ASC1 gene is regulated by the transcription factors Fhl1p and Ifh1p.
CPC2/ASC1 is one of the most abundantly transcribed genes in Saccharomyces cerevisiae. It encodes a ribosome-associated Gbeta-like WD protein, which is highly conserved from yeast to man. Here, we show that CPC2 transcription depends on the carbon source and is induced during utilization of the sugar glucose. CPC2 promoter deletion and insertion analyses identified two upstream activation sequence elements for CPC2, which are required for basal expression and regulation. One of these upstream activation sequence elements has an ATGTACGGATGT motif, which has previously been described as a putative binding site for the forkhead-like transcription factor Fhl1p. Deletion of FHL1 reduces CPC2 transcription significantly in presence of glucose, but has no effect when the non-fermentable carbon source ethanol is provided. Increased amounts of the Fhl1p co-regulator Ifh1p induce CPC2 transcription even when ethanol is utilized. These data suggest that the interaction between Fhl1p and Ifh1p is critical for the regulation of CPC2 transcription during utilization of different carbon sources.
Adaptor Proteins, Signal Transducing↗
Hydrolysis of lignocellulosic materials for ethanol production: a review.
Lignocellulosic biomass can be utilized to produce ethanol, a promising alternative energy source for the limited crude oil. There are mainly two processes involved in the conversion: hydrolysis of cellulose in the lignocellulosic biomass to produce reducing sugars, and fermentation of the sugars to ethanol. The cost of ethanol production from lignocellulosic materials is relatively high based on current technologies, and the main challenges are the low yield and high cost of the hydrolysis process. Considerable research efforts have been made to improve the hydrolysis of lignocellulosic materials. Pretreatment of lignocellulosic materials to remove lignin and hemicellulose can significantly enhance the hydrolysis of cellulose. Optimization of the cellulase enzymes and the enzyme loading can also improve the hydrolysis. Simultaneous saccharification and fermentation effectively removes glucose, which is an inhibitor to cellulase activity, thus increasing the yield and rate of cellulose hydrolysis.
Biomass↗
A quantitative electron microscopic study of synapse formation in dispersed cell cultures of rat cerebellum stained either by Os-UL or by E-PTA.
Synapse formation was followed in dispersed cell cultures of rat cerebellums stained either by osmium-uranyl-lead (Os-UL) or by ethanolic phosphotungstic acid (E-PTA). The numerical densities of synapses stained by Os-UL were always significantly higher than those stained by E-PTA (from 3 to 35 days in vitro). This difference suggests that some portion of the populations of both immature and mature synapses was not stained by E-PTA. The width of the synaptic cleft (28.4nm) in synapses stained by E-PTA was only 9nm more than that of the cleft in synapses stained by Os-UL (19.4nm), suggesting that some portion of one or both of the synaptic membranes is stained by E-PTA. Analysis of data from 7 other staining procedures utilizing both ethanolic and aqueous solutions demonstrated that the differences in cleft width described above appear to be due to the various affinities of the stains for different portions of synaptic membranes, and do not represent differences due to shrinkage artifact. In examining the parameters of synaptic structure during development of the cultures, a statistically significant increase in both the height and width of the presynaptic dense projections was found. Changes in synaptic morphology during synaptogenesis in this culture system were similar to those reported for the cerebellar cortex in vivo.
Animals↗
Hospital Use of Ethanol Survey (HUES): preliminary results.
Little information exists about alcohol use within health facilities. We sought to determine alcohol use and control in acute-care hospitals by mailing a questionnaire to a convenience sample of Pharmacy Directors of 24 hospitals in two regions. Of 23 responders, in-patient alcohol was dispensed by 21 (91%) within the last 5 years. Of these 21, both beverage and intravenous alcohol were dispensed by 13 (62%), only beverage alcohol by seven (33%), and only intravenous alcohol by one (5%). No institutional policies regarding alcohol dispensing existed in 16 (70%) hospitals. Alcohol was frequently used as a patient courtesy (14/20, 70%), and to prevent withdrawal (7/20, 35%). All pharmacies procured intravenous alcohol in a formal process, but 60% (12/20) obtained beverage alcohol informally. Alcohol is widely dispensed with few guidelines in this sample of acute-care hospitals. Additional research on therapeutic efficacy, consequences, and institutional oversight of alcohol in hospitals is needed.
Alcohol Withdrawal Delirium↗
As the occasion arises: PRN sedative orders in alcohol withdrawal treatment.
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Anti-Anxiety Agents↗
Determination of the relative ploidy in different Saccharomyces cerevisiae strains used for fermentation and 'flor' film ageing of dry sherry-type wines.
The full chromosomal karyotype of six enological Saccharomyces cerevisiae strains used for fermentation and biological ageing of sherry-type wines was studied. A genetic method based on the analysis of segregation frequencies of auxotrophic markers, among random spore progeny of hybrids, constructed between laboratory and industrial wine strains (Bakalinsky and Snow, 1990) was used. This method was combined with the analysis of strains by pulsed-field gel electrophoresis. The results obtained clearly indicate the presence of two, three or four copies of a chromosome in the industrial strains examined, and thus confirm that aneuploidy/polyploidy is not uncommon in these strains. In all strains examined, chromosome XIII polysomy is observed. This chromosome contains the ADH2 and ADH3 loci, that code for the ADHII and ADHIII isoenzymes of alcohol dehydrogenase, which are involved in ethanol oxidative utilization during biological ageing of wines. Tetrad analysis for the 'flor formation' character suggest two possibilities: this character is either regulated by at least a digenic system, or by only one gene present on a chromosome which is, at least, disomic.
Chromosomes, Fungal↗
Precise mass measurement of a double-stranded 500 base-pair (309 kDa) polymerase chain reaction product by negative ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.
Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS) has been used to determine the mass of a double-stranded 500 base-pair (bp) polymerase chain reaction (PCR) product with an average theoretical mass of the blunt-ended (i.e. unadenylated) species of 308 859.35 Da. The PCR product was generated from the linearized bacteriophage Lambda genome which is a double-stranded template. Utilization of ethanol precipitation in tandem with a rapid microdialysis step to purify and desalt the PCR product was crucial to obtain a precise mass measurement. The PCR product (0.8 pmol/µL) was electrosprayed from a solution containing 75% acetonitrile, 25 mM piperidine, and 25 mM imidazole and was infused at a rate of 200 nL/min. The average molecular mass and the corresponding precision were determined using the charge-states ranging from 172 to 235 net negative charges. The experimental mass and corresponding precision (reported as the 95% confidence interval of the mean) was 309 406 +/- 27 Da (87 ppm). The mass accuracy was compromised due to the fact that the PCR generates multiple products when using Taq polymerase due to the non-template directed 3'-adenylation. This results in a mixture of three PCR products with nearly identical mass (i.e. blunt-ended, mono-adenylated and di-adenylated) with unknown relative abundances that were not resolved in the spectrum. Thus, the experimental mass will be a weighted average of the three species which, under our experimental conditions, reflects a nearly equal concentration of the mono- and di-adenylated species. This report demonstrates that precise mass measurements of PCR products up to 309 kDa (500 bp) can be routinely obtained by ESI-FTICR requiring low femtomole amounts. Copyright 1999 John Wiley & Sons, Ltd.
Journal Article↗
Potentiometric method for substrate analysis using immobilized NAD + -dependent oxidoreductase enzymes.
Two coenzyme-dependent oxidoreductases, glucose dehydrogenase and alcohol dehydrogenase, were immobilized in polyacrylamide gel over a platinum grid matrix and used as enzyme electrodes to measure their substrate concentrations in buffered aqueous solutions. The immobilized enzymes were used to oxidize their substrates in the presence of NAD +. Ferricyanide was used as the redox mediator and electroactive species. The determinations of glucose and ethanol were utilized to demonstrate and evaluate the performance of the system. The described methodology should be readily applicable to the analysis of numerous other substrates of coenzyme-dependent oxidoreductases.
Acrylamides↗
Medication and fitness to drive.
PURPOSE: The aim of this study is to analyze the consumption patterns of medicaments among motor vehicle drivers who attend 'Medical Driving Test Centres' and the relation between habitual consumption of medicaments and fitness to drive. METHODS: The study was carried out on 8043 drivers who attended 25 Medical Driving Test Centres. RESULTS: 24.7% of drivers chronically consume medicaments while 6.8% consume medicaments along with alcohol every day. Of those who chronically consume medicaments with a warning about the medications on driving, 65.8% were considered 'fit' to drive, 27.3% 'fit with restrictions', 5.1% 'suspended' and 0.4% 'unfit'. CONCLUSIONS: The results show how frequent the consumption of medicaments along with alcohol is and that the great majority of drivers who take medicaments are considered fit to drive.
Adolescent↗
Expression of mammalian Rab Escort protein-1 and -2 in yeast Saccharomyces cerevisiae.
Rab GTPases are post-translationally geranylgeranylated on their C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase) and this modification is essential for their biological activity. Rab Escort Protein (REP) is a molecular chaperone that assists in the prenylation reaction carried out by RabGGTase. Mutations in the REP-1 gene lead to progressive retinal degradation and blindness in humans. Despite the significant interest in REP proteins, their preparative production remains challenging. We report here an inducible expression system for the production of REP-1 and REP-2 in yeast Saccharomyces cerevisiae at levels 3.5 and 2mg/L, respectively. The REP-1 was found to be toxic for yeast cells and its toxicity is proposed to be associated with the formation of unproductive Ypt:REP-1 complexes. To minimize the toxic effect of REP-1, the recombinant protein expression was induced at the end of the exponential phase. Under these conditions, the GAL1 promoter is no longer repressed due to exhaustion of glucose and utilization of ethanol as a carbon source. This expression procedure was successfully scaled up to 30L for both proteins. The REP-1 and REP-2 were purified using a combination of affinity and anion-exchange chromatography. Purified proteins were functionally active, as determined by a fluorescent Rab binding assay and in vitro prenylation. The reported procedure provides a reliable source of REP proteins for biochemical and structural studies.
Adaptor Proteins, Signal Transducing↗
Utilization of over-the-counter drugs during pregnancy.
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Analgesics↗
Methanoculleus chikugoensis sp. nov., a novel methanogenic archaeon isolated from paddy field soil in Japan, and DNA-DNA hybridization among Methanoculleus species.
A strictly anaerobic, irregularly coccoid, methanogenic archaeon, strain MG62T (= JCM 10825T = DSM 13459T), was isolated from paddy field soil in Chikugo, Fukuoka, Japan. The cells stained gram-negative, were 1.0-2.0 microm in diameter, were lysed by SDS and hypotonic solutions and were flagellated. Motility was not observed. The strain was able to use H2/CO2, 2-propanol/CO2, formate, 2-butanol/CO2 and cyclopentanol/CO2 as substrates for methanogenesis, but did not utilize acetate, ethanol, methanol or methylamines. The optimum temperature and pH were 25-30 degrees C and 6.7-7.2. Analysis of lipid component parts (core lipids, phospholipid polar head groups and glycolipid sugar moieties) showed the characteristic pattern of members of the family Methanomicrobiaceae except for the absence of glucose as a glycolipid sugar moiety. The G+C content of the DNA was 62.2 mol %. Sequence analysis of the 16S rDNA revealed that the strain belonged to the genus Methanoculleus. The strain had DNA-DNA hybridization values of less than 50% with type strains of Methanoculleus species. On the basis of phenotypic, genotypic and phylogenetic characteristics, the name Methanoculleus chikugoensis sp. nov. is proposed for strain MG62T (= JCM 10825T = DSM 13459T). The DNA hybridization study also revealed the close relationships of three species, Methanoculleus olentangyi, Methanoculleus bourgensis and Methanoculleus oldenburgensis, among Methanoculleus species.
Base Composition↗