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Immunomodulatory effect of a polysaccharide-enriched preparation of Codonopsis pilosula roots.

1. A polysaccharide-enriched fraction (CPPS) was prepared from Codonopsis pilosula root extract utilizing a procedure that entailed extraction with aqueous buffer and precipitation with ethanol. 2. After administration of CPPS in drinking water to C57BL/6 mice at a dosage of 10 mg/L for 4 weeks, the splenocytes exhibited lowered mitogenic responses to Concanavalin A (ConA) and lipopolysaccharide (LPS). The in vitro production of reactive nitrogen intermediates was inhibited. 3. However, when oral administration of CPPS was prolonged to 8 weeks, there was a potentiation of ConA-stimulated and LPS-stimulated mitogenic responses. 4. When tested under in vitro conditions, CPPS augmented the mitogenic response of splenocytes to ConA. However, there was no effect on the pinocytic activity of mouse macrophages, nor was there any proliferative activity on mouse melanoma B16 cells.

Adjuvants, Immunologic↗

Interactions between propionate and amino acid metabolism in isolated sheep hepatocytes.

The purpose of the present study was to evaluate the contribution of various substrates to glucose synthesis in isolated sheep hepatocytes, and more specifically to quantify the contribution of propionate to gluconeogenesis. Liver cells from fed sheep have a very high capacity for propionate utilization and conversion into glucose. The gluogenicity of lactate or amino acids was very low in hepatocytes from fed sheep, but was significantly increased in hepatocytes from starved animals. Amino acids such as alanine or glutamine were characterized by a substantial utilization towards ureogenesis; whereas their conversion to glucose was very low. Propionate utilization and conversion into glucose was inhibited by butyrate, ammonia and especially ethanol (by up to 80%). Ethanol promoted a striking accumulation of intracellular malate in hepatocytes incubated with propionate (reaching 14.9 mumol/g cell) and led to a depletion of phosphoenolpyruvate; ethanol inhibition could be counteracted by pyruvate. Propionate and butyrate enhanced ureogenesis from ammonia in ruminant liver cells but their effects were not additive. Propionate also elicited a marked increase in cellular concentrations of phosphoserine and serine, particularly in the presence of ammonia; such effects could influence phospholipid metabolism in the liver. These findings emphasize the contribution of propionate, compared with the other glucogenic substrates, to glucose synthesis in ruminants and point to the possibilities of modulation of the glucogenicity of propionate by various substrates which may be present in portal blood.

Amino Acids↗

The effects of ethanol on the developing cerebellum and eyeblink classical conditioning.

In rats, developmental ethanol exposure has been used to model the central nervous system deficits associated with human fetal alcohol syndrome. Binge-like ethanol exposure of neonatal rats depletes cells in the cerebellum, including Purkinje cells, granule cells, and deep nuclear cells, and produces deficits in simple tests of motor coordination. However, the extent to which anatomical damage is related to behavioral deficits has been difficult to estimate. Eyeblink classical conditioning is known to engage a discrete brain stem-cerebellar circuit, making it an ideal test of cerebellar functional integrity after developmental ethanol exposure. Eyeblink conditioning is a simple form of motor learning in which a neutral stimulus (such as a tone) comes to elicit an eyeblink when repeatedly paired with a stimulus that evokes an eyeblink prior to training (such as mild periorbital stimulation). In eyeblink conditioning, one of the deep cerebellar nuclei, the interpositus nucleus, as well as specific Purkinje cell populations, are sites of convergence for tone conditioned stimulus and somatosensory unconditioned stimulus information, and, together with brain stem nuclei, provide the necessary and sufficient substrate for the learned response. A series of studies have shown that eyeblink conditioning is impaired in both weanling and adult rats given binge-like exposure to ethanol as neonates. In addition, interpositus nucleus neurons from ethanol-exposed rats showed impaired activation during eyeblink conditioning. These deficits are accompanied by a permanent reduction In the deep cerebellar nuclear cell population. Because particular cerebellar cell populations are utilized in well-defined ways during eyeblink conditioning, conclusions regarding the underlying neural substrates of behavioral change after developmental ethanol exposure are greatly strengthened.

Animals↗

Serum-ethanol determination: comparison of lactate and lactate dehydrogenase interference in three enzymatic assays.

Gas chromatography is considered to be the reference method for ethyl alcohol determination. However, enzymatic ethanol assays have been developed for use in the clinical laboratory by several commercial vendors. Essentially, these assays utilize the oxidation of ethyl alcohol to acetaldehyde with concurrent reduction of nicotinamide adenine dinucleotide (NAD) to NADH while monitoring the increase in absorbance at 340 nm. The increase in absorbance is theoretically proportional to the ethanol concentration in the sample. Previously, several authors reported that increased concentrations of lactate and lactate dehydrogenase (LDH) can cause false-positive results with certain enzymatic ethyl alcohol assays. In the present investigation, we further studied the interference of lactate and LDH in three enzymatic assays. Apparent ethyl alcohol concentrations in serum spiked with lactate and LDH, as well as patient and autopsy samples, were determined by the Syva, Abbott, and Roche enzymatic assays and by gas chromatography. The effect of coenzyme depletion on the rate of reaction and the interference of hemolysis were also investigated. Based on our results we suggest that coenzyme depletion plays a major role in the severity of the false-positive ethyl alcohol result, and the interference from hemolysis has a negligible effect on these results. We also confirm the previous studies in showing that elevated serum-lactate and LDH concentrations can result in varying degrees of false-positive ethyl alcohol concentrations in the three enzymatic assays. This should be taken into consideration in the management of patients in a tertiary care medical center.

Catalysis↗

Desulfotomaculum carboxydivorans sp. nov., a novel sulfate-reducing bacterium capable of growth at 100% CO.

A moderately thermophilic, anaerobic, chemolithoheterotrophic, sulfate-reducing bacterium, strain CO-1-SRB(T), was isolated from sludge from an anaerobic bioreactor treating paper mill wastewater. Cells were Gram-positive, motile, spore-forming rods. The temperature range for growth was 30-68 degrees C, with an optimum at 55 degrees C. The NaCl concentration range for growth was 0-17 g l(-1); there was no change in growth rate until the NaCl concentration reached 8 g l(-1). The pH range for growth was 6.0-8.0, with an optimum of 6.8-7.2. The bacterium could grow with 100% CO in the gas phase. With sulfate, CO was converted to H(2) and CO(2) and part of the H(2) was used for sulfate reduction; without sulfate, CO was completely converted to H(2) and CO(2). With sulfate, strain CO-1-SRB(T) utilized H(2)/CO(2), pyruvate, glucose, fructose, maltose, lactate, serine, alanine, ethanol and glycerol. The strain fermented pyruvate, lactate, glucose and fructose. Yeast extract was necessary for growth. Sulfate, thiosulfate and sulfite were used as electron acceptors, whereas elemental sulfur and nitrate were not. A phylogenetic analysis of 16S rRNA gene sequences placed strain CO-1-SRB(T) in the genus Desulfotomaculum, closely resembling Desulfotomaculum nigrificans DSM 574(T) and Desulfotomaculum sp. RHT-3 (99 and 100% similarity, respectively). However, the latter strains were completely inhibited above 20 and 50% CO in the gas phase, respectively, and were unable to ferment CO, lactate or glucose in the absence of sulfate. DNA-DNA hybridization of strain CO-1-SRB(T) with D. nigrificans and Desulfotomaculum sp. RHT-3 showed 53 and 60% relatedness, respectively. On the basis of phylogenetic and physiological features, it is suggested that strain CO-1-SRB(T) represents a novel species within the genus Desulfotomaculum, for which the name Desulfotomaculum carboxydivorans is proposed. This is the first description of a sulfate-reducing micro-organism that is capable of growth under an atmosphere of pure CO with and without sulfate. The type strain is CO-1-SRB(T) (=DSM 14880(T)=VKM B-2319(T)).

Anaerobiosis↗

The epidemiology of the homeless population and its impact on an urban emergency department.

OBJECTIVES: To characterize the homeless adult population of an urban emergency department (ED) and study the medical, psychiatric, and social factors that contribute to homelessness. METHODS: A prospective, case-control survey of all homeless adult patients presenting to an urban, tertiary care ED and a random set of non-homeless controls over an eight-week period during summer 1999. Research assistants administered a 50-item questionnaire and were trained in assessing dentition and triceps skin-fold thickness. INCLUSION CRITERIA: all homeless adults who consented to participate. Homelessness was defined as being present for any person not residing at a private address, group home, or drug treatment program. Randomly selected controls were concurrently enrolled with a 3:1 homeless:control rate. EXCLUSION CRITERIA: critically ill, injured, or incapacitated patients, or patients <21 years of age. Univariate analysis with appropriate statistical tests was used. The Mantel-Haenszel test was used to adjust for population differences. RESULTS: Two hundred fifty-two homeless subjects and 88 controls were enrolled. Data are presented for homeless vs control patients, and all p-values were <0.01. Odds ratios (ORs) with 95% confidence intervals (95% CIs) are given where appropriate: mean age (+/-SD) = 42 +/- 10 vs 48 +/- 13; male gender 95% vs 54% (OR = 17; 95% CI = 8 to 37); history of (hx) tuberculosis 49% vs 15% (OR = 2.5; 95% CI = 1.2 to 3); hx HIV infection 35% vs 13% (OR = 3.8; 95% CI = 1.8 to 8); hx penetrating trauma 62% vs 16% (OR = 8.62; 95% CI = 4.4 to 17.1); hx depression 70% vs 15% (OR = 13.4; 95% CI = 6.7 to 27); hx schizophrenia 27% vs 7% (OR = 5.1; 95% CI = 2.0 to 14); hx alcoholism 81% vs 15% (OR = 24; 95% CI = 12 to 49); significant tooth loss (>3) 43% vs 18% (OR = 3.3; 95% CI = 1.8 to 6.4); percentage of body fat 16.5% vs 19.7%; hx social isolation (no weekly social contacts) 81% vs 11% (OR = 33.3; 95% CI = 14 to 100); mean number of ED visits/year 6.0 vs 1.6. CONCLUSIONS: In the study population homelessness was associated with a history of significantly higher rates of infectious disease, ethanol and substance use, psychiatric illness, social isolation, and rates of ED utilization.

Adult↗

Expression of a Lactose Transposon (Tn951) in Zymomonas mobilis.

The potential utility of Zymomonas mobilis as an organism for the commercial production of ethanol would be greatly enhanced by the addition of foreign genes which expand its range of fermentable substrates. We tested various plasmids and mobilizing factors for their ability to act as vectors and introduce foreign genes into Z. mobilis CP4. Plasmid pGC91.14, a derivative of RP1, was found to be transferred from Escherichia coli to Z. mobilis at a higher frequency than previously reported for any other plasmids. Both tetracycline resistance and the lactose operon from this plasmid were expressed in Z. mobilis CP4. Plasmid pGC91.14 was stably maintained in Z. mobilis at 30 degrees C but rapidly lost at 37 degrees C.

Journal Article↗

Development of Microbodies in the yeast Kloeckera growing on methanol.

A number of microbodies appear regularly in methanol-grown yeast cells, but rarely in ethanol- or glucose-grown cells. When one of representative methanol-utilizing yeasts, Kloeckera sp.no. 2201 (also known as Candida bodinii), was cultured on glucose and then transferred into a methanol medium, microbodies of small size could be observed in 2-h old cells. The number of microbodies per sectioned cell reached five to six after 4 h of cultivation. Though the number of microbodies did not change during prolonged cultivation, their size became larger with the passage of cultivation time. The activities of catalase and alcohol oxidase were confirmed in the particulate fractions throughout the cultivation period, whereas the activities of formaldehyde dehydrogenase and formate dehydrogenase were not detected in the particles. The activity of isocitrate lyase was detected in the particulate fractions only at the early growth phase.

Alcohol Oxidoreductases↗

Uptake of monohydric alcohols by liver: demonstration of a shared enzymic space.

Multiple-indicator dilution studies of the hepatic uptake of straight-chain C1-C5 monohydric alcohols were carried out in anesthetized dogs, with either no preceding or saturating infusions of ethanol, and at different steady-state levels for the C2 experiments. Labeled red cells were utilized as a vascular reference, and labeled water was used as a second reference entering liver cells. Kinetic analysis of the data provided estimates of both an uptake rate constant and the space of distribution available to label. From the decrease in the uptake rate constant for labeled ethanol with bulk concentration, we calculated a maximal removal rate of 0.025 mumol X s-1 X (ml liver water)-1 and a Michaelis constant (Km) of 0.32 mM. Especially for the labeled C3-C5 alcohols, a space in excess of that available to water was found, and the bulk of this was dissipated by ethanol infusion. The increment, the "shared enzymic space", which varies with enzymic concentration and inversely with Km, was used to calculate Km values for the other alcohols.

Alcohol Dehydrogenase↗

Dietary influences on alcohol intake: a review.

Alcohol intake is affected by both environmental and inherited biological mechanisms. In the early history of alcohol research, caloric interactions were among the most intensely studied of the environmental factors, and a large body of evidence was obtained that indicated that nutrition can affect alcohol drinking. Much of this evidence still stands today but appears to have been largely overlooked and to be in danger of being forgotten. Nevertheless, alcohol is not only a pharmacological substance but also a nutrient. As a nutrient it influences the metabolism of most tissues in the body, with especially marked effects on glucose homeostasis. Alcohol has a high energy content, and this energy is utilized by the body as efficiently as the energy in normal food. Ethanol has such good properties as a substrate for energy production that we are faced with the problem of explaining, not why it is consumed, but why it is not consumed in still larger quantities by nonalcoholic humans or by animals. When alcohol is consumed by animals, the intake of food decreases in relation to the caloric content of the alcohol; if a choice of macronutrients is possible, alcohol decreases the consumption of carbohydrates most. The interaction between alcohol and food intake goes both ways, however, with the intake of different foods also influencing alcohol consumption. For example, a high carbohydrate/low protein diet depresses alcohol intake, whereas a low carbohydrate/high protein food increases it. If such specific diets can help to depress alcohol intake, nutritional therapy might be useful in the treatment of alcohol abusers, probably not as the primary treatment, but perhaps as an adjunct to other forms of treatment.

Alcohol Drinking↗

[Geoalkalibacter ferrihydriticus gen. nov., sp. nov., the first alkaliphilic representative of the family Geobacteraceae, isolated from a soda lake].

Investigation of iron reduction in bottom sediments of alkaline soda lakes resulted in the isolation of a new obligately anaerobic iron-reducing bacterium, strain Z-0531, from Lake Khadyn (Tuva Republic, Russia) sediment samples. The cells of strain Z-0531 are short (1.0-1.5 by 0.3-0.5 microm), motile, non-spore-forming, gram-negative rods. The isolate is an obligate alkaliphile, developing in the pH range of 7.8-10.0, with an optimum at pH 8.6. It does not require NaCl but grows at NaCl concentrations of 0-50 g/1l. It can oxidize acetate with such electron acceptors as amorphous Fe(llI) hydroxide (AFH), EDTA-Fe(III), anthraquinone-2,6-disulfonate (quinone), Mn(IV), and S(0). On media with EDTA-Fe(III), the isolate can oxidize, apart from acetate, ethanol, pyruvate, oxalate, arginine, tartrate, lactate, propionate, and serine. H2 is not utilized. The reduced products formed during growth with AFH are siderite or magnetite, depending on the growth conditions. The isolate is incapable of fermenting sugars, peptides, and amino acids. Yeast extract or vitamins are required as growth factors. The organism is capable of dinitrogen fixation and harbors the nifH gene. The DNA G+C content is 55.3 mol %. 16S rRNA analysis places strain Z-0531 into the family Geobacteraceae. Its closest relative (93% similarity) is Desulfuromonas palmitatis. Based on phenotypic distinctions and phylogenetic position, it is proposed that strain Z-0531 be assigned to the new genus and species Geoalkalibacter ferrihydriticus gen. nov., sp. nov.

Acetates↗

Drug impairment reviews: other drugs.

This review will summarize a limited number of experimental studies concerning effects of tranquilizers, muscle relaxants, and antihistamine drugs, as well as combinations of these drugs with ethanol, on human performance related to driving. The first group of studies utilized batteries of psychological and/or psychomotor tests to assess drug effects, while the second group used driving simulators. Procedures and results of each study will be summarized, and this will be followed by a general critique of methodologies which have been used along with suggestions for improved methodologies.

Automobile Driving↗

[Functional characteristics of the electron transport chain in Candida mycoderma yeasts exposed to benzhydroxamic acid].

The effect of antimycin A and benzhydroxamic acid (BHA) on functioning of the electron transport system was studied with the resting cells of Candida mycoderma grown in a medium containing glucose and collected at the beginning of the deceleration phase. In the original ("control") cells, the processes of oxygen consumption were shown to be mediated mainly by the phosphorylating electron transport chain. When the cells were incubated withe glucose, the cyanide resistant electron transport chain (CRETC) started to operate in these processes. Antimycin A stimulated the operation of CRETC. BHA blocked CRETC and, at the same time, prevented the participation of pyridine nucleotides (PN) in the processes of oxygen utilization. Apparently, electron donors (or reduced equivalents) in the electron transport chain were flavin enzymes operating in the cytochrome b region of the chain instead of PN. Similar changes in the electron transport system (when PN were not involved in the processes of oxygen utilization) were observed if resting cells were incubated in the presence of ethanol.

Antimycin A↗

Modeling the synchronization of yeast respiratory oscillations.

The budding yeast Saccharomyces cerevisiae exhibits autonomous oscillations when grown aerobically in continuous culture with ethanol as the primary carbon source. A single cell model that includes the sulfate assimilation and ethanol degradation pathways recently has been developed to study these respiratory oscillations. We utilize an extended version of this single cell model to construct large cell ensembles for investigation of a proposed synchronization mechanism involving hydrogen sulfide. Ensembles with as many as 10,000 cells are used to simulate population synchronization and to compute transient number distributions from asynchronous initial cell states. Random perturbations in intracellular kinetic parameters are introduced to study the synchronization of single cells with small variations in their unsynchronized oscillation periods. The cell population model is shown to be consistent with available experimental data and to provide insights into the regulatory mechanisms responsible for the synchronization of yeast metabolic oscillations.

Cell Culture Techniques↗

Selected mouse lines, alcohol and behavior.

The technique of selective breeding has been employed to develop a number of mouse lines differing in genetic sensitivity to specific effects of ethanol. Genetic animal models for sensitivity to the hypnotic, thermoregulatory, excitatory, and dependence-producing effects of alcohol have been developed. These genetic animal models have been utilized in numerous studies to assess the bases for those genetic differences, and to determine the specific neurochemical and neurophysiological bases for ethanol's actions. Work with these lines has challenged some long-held beliefs about ethanol's mechanisms of action. For example, lines genetically sensitive to one effect of ethanol are not necessarily sensitive to others, which demonstrates that no single set of genes modulates all ethanol effects. LS mice, selected for sensitivity to ethanol anesthesia, are not similarly sensitive to all anesthetic drugs, which demonstrates that all such drugs cannot have a common mechanism of action. On the other hand, WSP mice, genetically susceptible to the development of severe ethanol withdrawal, show a similar predisposition to diazepam and phenobarbital withdrawal, which suggests that there may be a common set of genes underlying drug dependencies. Studies with these models have also revealed important new directions for future mechanism-oriented research. Several studies implicate brain gamma-aminobutyric acid and dopamine systems as potentially important mediators of susceptibility to alcohol intoxication. The stability of the genetic animal models across laboratories and generations will continue to increase their power as analytic tools.

Animals↗

[Exopolysaccharide production and peculiarities of C6-metabolism in Acinetobacter sp. grown on carbohydrate substrates].

An Acinetobacter sp. strain grown on carbohydrate substrates (mono- and disaccharides, molasses, starch) was shown to synthesize exopolysaccharides (EPS). Glucose catabolism proved to proceed via the Embden-Meyerhof-Parnas and Entner-Doudoroff pathways. Pyruvate entered the tricarboxylic acid cycle due to pyruvate dehydrogenase activity. Pyruvate carboxylation by pyruvate carboxylase was the anaplerotic reaction providing for the synthesis of intermediates for the constructive metabolism of Acinetobacter sp. grown on C6-substrates. The C6-metabolism in Acinetobacter sp. was limited by coenzyme A. Irrespective of the carbohydrate growth substrate (glucose, ethanol), the activities of the key enzymes of both C2- and C6-metabolism was high, except for the isocitrate lyase activity in glucose-grown bacteria. Isocitrate lyase activity was induced by C2-compounds (ethanol or acetate). After their addition to glucose-containing medium, both substrates were utilized simultaneously, and an increase was observed in the EPS synthesis, as well as in the EPS yield relative to biomass. The mechanisms responsible for enhancing the EPS synthesis in Acinetobacter sp. grown on a mixture of C2- and C6-substrates are discussed.

Acetates↗

The discriminative stimulus properties of ethanol and acute ethanol withdrawal states in rats.

Twelve male Sprague-Dawley rats were trained in a standard two-choice Drug 1-Drug 2 discrimination task utilizing 3.0 mg/kg chlordiazepoxide (CDP, an anxiolytic drug) and 20 mg/kg pentylenetetrazol (PTZ, an anxiogenic drug) as discriminative stimuli under a VR 5-15 schedule of food reinforcement. Saline tests conducted at specific time points after acute high doses of ethanol (3.0 and 4.0 g/kg) indicated a delayed rebound effect, evidenced by a shift to PTZ-appropriate responding. Insofar as such a shift in lever selection indexes a delayed anxiety-like state, this acute 'withdrawal' reaction can be said to induce an affective state similar to that seen with chronic ethanol withdrawal states. Ethanol generalization tests: (1) resulted in a dose- and time-dependent biphasic generalization to CDP, (2) failed to block the PTZ stimulus and (3) failed to block the time- and dose-dependent elicitation of an ethanol-rebound effect. These data suggest that ethanol's anxiolytic effects are tenuous.

Affect↗