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PubMed1982

Extraction procedures for oilseeds and related high fat-low moisture products.

A combined sample preparation/extraction procedure is presented for pesticide residue analysis of oilseeds and related high fat-low moisture products. The procedure utilizes high-speed milling to prepare the sample and high-speed homogenization in the extraction step to achieve what is apparently quantitative isolation of both incurred residues and natural oils. A separate, simple, oil determination step allows findings to be reported on either the fat or whole product basis. Petroleum ether, ethyl ether-petroleum ether (1 + 1), and ethanol are used serially as the extractants. Usual fatty food cleanup procedures and multiresidue gas chromatographic detection techniques are utilized. The procedure presented in this paper is a refinement of earlier work which used a homogenizer both to grind and to extract samples of unground seeds and which demonstrated essentially complete extraction of endrin residues in soybeans and DDT residues in mustard seed. Identical samples analyzed by the currently recommended shakeout procedure, 29.012, gave recoveries of approximately 50% of the total residues. The procedure presented in this paper was satisfactorily tested on 13 different oilseed types and one sample of soda crackers. Oil content for these samples ranged from 5 to 69%.

Arachis↗
PubMed2006

Effect of ethanol on human osteosarcoma cell proliferation, differentiation and mineralization.

The habitual consumption of even moderate quantities of alcoholic beverages is clearly associated with reduced bone mass, increased prevalence of skeletal fracture and also it is the major risk factor for the development of secondary osteoporosis. The present in vitro study was designed to determine the dose response effects of ethanol on osteoblast-like human osteosarcoma cells (SaOS-2) proliferation, differentiation, mineralization and cyto-toxicity. SaOS-2 cells were plated in 48 and 6 well culture plates and exposed to different concentrations of ethanol (1, 10, 100, 200 and 300 mM) for 24, 48 and 72 h. At the end of incubation, proliferation of cells was studied using crystal violet Bioassay. The cell lysate was utilized to determine ALP activity and conditioned media were used to measure LDH activity. Histochemical localization of ALP and mineralized nodules were studied from cells treated with ethanol (10 and 100 mM) for 21 days. At higher doses, there was a significant reduction in cell number, whereas at lower doses there were variable effects. In 24 h treatment, the higher doses showed a significant increase in ALP activity, whereas 48 and 72 h treatments showed an opposite trend. Ethanol treatment caused a dose- and time-dependent increase in LDH activity. Ethanol treatment altered the quality of mineralization at 10 mM dose whereas completely inhibited mineralization at 100 mM dose, despite the presence of serum. In conclusion, the toxic effect of ethanol is reflected on cell proliferation, differentiation and mineralization even at low doses and at extended treatment duration.

Alkaline Phosphatase↗
PubMed1988

Acute effects of ethanol and acetate on glucose kinetics in normal subjects.

We compared the effects of two ethanol doses on glucose kinetics and assessed the role of acetate as a mediator of ethanol-induced insulin resistance. Ten normal males were studied on four occasions, during which either a low (blood ethanol 4 +/- 1 mmol/l) or moderate (14 +/- 1 mmol/l) ethanol, acetate, or saline dose was administered. Both ethanol doses similarly inhibited (0.4-0.5 mg.kg-1.min-1, P less than 0.01) basal glucose production. The decrease in Ra was matched by a comparable decrease in glucose utilization (Rd), resulting in maintenance of normoglycemia. During hyperinsulinemia (approximately 70 microU/ml), glucose disposal was lower (1.2-1.7 mg.kg-1.min-1, P less than 0.01) in the moderate than the low-dose ethanol or saline studies. During acetate infusion, the blood acetate level was comparable with those in the ethanol studies. Acetate had no effect on glucose kinetics. In conclusion, 1) in overnight fasted subjects, ethanol does not cause hypoglycemia because its inhibitory effect on Ra is counterbalanced by equal inhibition of Rd;2) basal Ra and Rd are maximally inhibited already by small ethanol doses, whereas inhibition of insulin-stimulated glucose disposal requires a moderate ethanol dose; and 3) acetate is not the mediator of ethanol-induced insulin resistance.

Acetates↗
PubMedDate not supplied

Intragastric ethanol infusion model for cellular and molecular studies of alcoholic liver disease.

The intragastric alcohol infusion rat model (IAIRM) of alcoholic liver disease (ALD) has been utilized in various laboratories to study various aspects of ALD pathogenesis including oxidative stress, cytokine upregulation, hypoxic damage, apoptosis, ubiquitin-proteasome pathway and CYP2E1 induction. The basic value of the model is that it produces pathologic changes which resemble ALD including microvesicular and macrovesicular fat, megamitochondria, apoptosis, central lobular and pericellular fibrosis, portal fibrosis, bridging fibrosis, central necrosis, and mixed inflammatory infiltrate including PMNs and lymphocytes. The model is valuable because the diet and ethanol intake are totally under the control of the investigator. A steady state can be maintained with high or low blood alcohol levels for long periods. The cycling of the blood alcohol levels, when a constant infusion rate of alcohol is maintained, simulates binge drinking. Using this model the importance of dietary fat, especially the degree of saturation of the fatty acids on the induction of liver pathology, has been documented. The role of endotoxin, the Kupffer cell, TNFalpha, and NADPH oxidase have been demonstrated. The importance of 2E1 in oxidative stress induction has been shown using inhibitors of the isozyme. The importance of dietary iron in the pathogenesis of cirrhosis has been documented. Acetaldehyde has been shown to play a role in preventing liver pathology by preventing NFkappaB activation. Using the model, to maintain high blood alcohol levels is found to be necessary to demonstrate proteasomal peptidase inhibition. Ubiquitin synthesis is also inhibited at high blood alcohol levels in the IAIRM model. Oxidized proteins accumulate in the liver at high blood alcohol levels. Neoantigens derived from protein adducts formed with products of oxidation induce autoimmune mechanisms of liver injury. Thus, in many ways the model has revolutionized our understanding of the pathogenesis of ALD.

Animals↗
PubMed1984

Excess free energy approach to the estimation of solubility in mixed solvent systems II: Ethanol-water mixtures.

The use of the reduced three-suffix solubility equation in characterizing solubility in ethanol-water mixtures is discussed. The equation states that ln xs2,m = z1 ln xs2,1 + z3 ln xs2,3 - A1-3z1z3(2z1 - 1)- (q2/q1) + A3-12z21z3(q2/q3) + C2z1z3, where xs2,1, xs2,3, and xs2,m are the mole fraction solubilities of the solute in ethanol (subscript 1), water (subscript 3), and in the mixture (m); A1-3 and A3-1 are solvent-solvent interaction terms; C2 is a solute-solvent interaction term; and the q- and z-values are molar volumes and solute-free volume fractions, respectively. The contributions of the various terms in the equation to solubility are examined, and the possible use of its derivative in indicating whether a maximum may exist in the solubility profile is discussed. Methods of obtaining the solvent-solvent interaction constant and the ternary constant C2 are described, and the general effectiveness of the equation in describing solubility is examined. The equation is shown to be applicable to 10 compounds with widely different physical properties and, thus, appears to combine both ease of use and general utility.

Chemical Phenomena↗
PubMedDate not supplied

Process considerations and economic evaluation of two-step steam pretreatment for production of fuel ethanol from softwood.

To increase the overall ethanol yield from softwood, the steam pretreatment stage can be carried out in two steps. The two-step pretreatment process was evaluated from a techno-economic standpoint and compared with the one-step pretreatment process. The production plants considered were designed to utilize spruce as raw material and have a capacity of 200,000 tons/year. The two-step process resulted in a higher ethanol yield and a lower requirement for enzymes. However, the two-step process is more capital-intensive and has a higher energy requirement. The estimated ethanol production cost was the same, 4.13 SEK/L (55.1 cent /L) for both alternatives. For the two-step process different energy-saving options were considered, such as a higher concentration of water-insoluble solids in the filter cake before the second step, and the possibility of excluding the pressure reduction between the steps. The most optimistic configuration, with 50% water-insoluble solids in the filter cake in the feed to the second pretreatment step, no pressure reduction between the pretreatment steps, and 77% overall ethanol yield (0.25 kg EtOH/kg dry wood), resulted in a production cost of 3.90 SEK/L (52.0 cent /L). This shows the potential for the two-step pretreatment process, which, however, remains to be verified in pilot trials.

Bioreactors↗
PubMed2003

Sterol glucosyltransferases have different functional roles in Pichia pastoris and Yarrowia lipolytica.

Mutants of the methanol-utilizing yeast Pichia pastoris and the alkane-utilizing yeast Yarrowia lipolytica defective in the orthologue of UGT51 (encoding sterol glucosyltransferase) were isolated and compared. These mutants do not contain the specific ergosterol derivate, ergosterol glucoside. We observed that the P. pastoris UGT51 gene is required for pexophagy, the process by which peroxisomes containing methanol-metabolizing enzymes are selectively shipped to and degraded in the vacuole upon shifting methanol-grown cells of this yeast to glucose or ethanol. PpUGT51 is also required for other vacuole related processes. In contrast, the Y. lipolytica UGT51 gene is required for utilization of decane, but not for pexophagy. Thus, sterol glucosyltransferases play different functional roles in P. pastoris and Y. lipolytica.

Alkanes↗
PubMed2004

The deletion of the succinate dehydrogenase gene KlSDH1 in Kluyveromyces lactis does not lead to respiratory deficiency.

We have isolated a Kluyveromyces lactis mutant unable to grow on all respiratory carbon sources with the exception of lactate. Functional complementation of this mutant led to the isolation of KlSDH1, the gene encoding the flavoprotein subunit of the succinate dehydrogenase (SDH) complex, which is essential for the aerobic utilization of carbon sources. Despite the high sequence conservation of the SDH genes in Saccharomyces cerevisiae and K. lactis, they do not have the same relevance in the metabolism of the two yeasts. In fact, unlike SDH1, KlSDH1 was highly expressed under both fermentative and nonfermentative conditions. In addition to this, but in contrast with S. cerevisiae, K. lactis strains lacking KlSDH1 were still able to grow in the presence of lactate. In these mutants, oxygen consumption was one-eighth that of the wild type in the presence of lactate and was normal with glucose and ethanol, indicating that the respiratory chain was fully functional. Northern analysis suggested that alternative pathway(s), which involves pyruvate decarboxylase and the glyoxylate cycle, could overcome the absence of SDH and allow (i) lactate utilization and (ii) the accumulation of succinate instead of ethanol during growth on glucose.

Amino Acid Sequence↗
PubMed1982

[Method of plasma acetate determination by gas chromatography after ethanol administration in man (author's transl)].

Plasma acetate is extracted using ethanol in alkaline medium, in presence of internal standard. After concentration of the hydroalcoholic phase, the chromatographic analysis is performed utilizing 10% SP 1200 and 1% H3PO4 on chromosorb W. The linearity of the method has been tested from 0.05 to 10 mM/I, its reproducibility is +/- 3.5% for levels ranging from 0.1 to 10 mM/I and +/- 10% for those smaller than 0.1 mM/I. This method is sensitive enough to assess the physiological acetate levels and those obtained after ethanol biotransformation.

Acetates↗
PubMed1997

Evaluation of different solvent systems for the extraction and fractionation of oleoresins from guajillo peppers.

Dried guajillo peppers were first extracted with four different solvents: ethanol, acetone, ethyl acetate and hexane with the aim of obtaining oleoresins which were further fractionated into red and paprika extracts. Results showed that as the polarity of the solvent increased the amount of pigments extracted also increased. Acetone had good affinity for pungent (capsaicin) compounds. Utilization of these solvents alone did not produce red and paprika oleoresins that meet commercial specifications. Fractionation of acetone extracted oleoresins with ethanol: water (90:10) yielded a precipitate and a solution. The precipitate and solution produced red and paprika extracts that meet pungency and color specifications. It was possible to obtain red and paprika oleoresins from mild guajillo peppers.

Capsicum↗
PubMed1978

Effects of various metabolites (sugars, carboxylic acids and alcohols) on riboflavin formation in non-growing cells of Ashbya gossypii.

The effects of various sugars and sugar derivatives on flavinogenesis were examined using non-growing cells of a high flavinogenic mold, Ashbya gossypii. Glucose, fructose and galactose were found to be the most stimulative. Glycerol and glucono-delta-lactone were less stimulative; next in order were n-propanol, n-butanol, glycols and butanediols, which were likewise effective; acetate, lactate and pyruvate were slightly stimulative. In contrast, ribose, xylose, arabinose, ribitol, citrate, succinate, oxaloacetate, glyoxylate and malate were rather inhibitory, in additions at 1.0%. Among these compounds, ethanol (1%) greatly stimulated riboflavin formation. Maximum flavinogenesis with the above stimulants was attained by the additions of 1% ethanol, 1.25--3.0% glucose, 1.25% glycerol, 4.0--6.0% propane and butanediols, 1.0% pyruvate and 0.9% acetate after 37 hr incubation, respectively. These compounds inhibited flavinogenesis with increasing concentrations above their optimum concentrations. The stimulation effect of ethanol far exceeded those of other stimulants but ethanol had almost no effect on growth and pH values during incubation. With the addition of ethanol (1%) during incubation, maximum formation (1,776 microgram/g wet mycelia) of riboflavin was achieved when added at the start of incubation and the most effective utilization was observed when added at the logarithmic phase of flavinogenesis, although the maximum formation of riboflavin in the latter case was much lower than in the former case. The relation of sugar metabolism, especially ethanol metabolism, to flavinogenesis was discussed with the flavinogenic activities of these additives.

Acetates↗
PubMedDate not supplied

Pup-induced maternal behavior in adult and juvenile rats exposed to alcohol prenatally.

The effects of prenatal alcohol exposure on pup-induced maternal behaviors were examined in virgin rats whose mothers consumed liquid diets containing either 35 or 0% ethanol-derived-calories (EDC) on days 6-20 of gestation. A pair-feeding procedure was utilized to control for reduced caloric intake and, in addition, an ad libitum lab chow group was included to control for possible liquid diet effects. The results from experiments 1 and 2 revealed a sexually differential effect on this behavior in adult virgins exposed to alcohol in utero. Adult 35% EDC females showed a variety of deficits in maternal behaviors relative to controls; however, males with similar prenatal treatment appeared unaffected. In experiment 3, both 35% EDC male and female juveniles displayed altered maternal behaviors relative to controls. These results demonstrate yet another behavior sensitive to prenatal alcohol exposure. Prenatal hormonal alterations occurring as a consequence of maternal alcohol consumption may be a mechanism by which this long term behavioral effect is observed. This hypothesis and the importance of future research in this relatively new area of fetal alcohol studies are discussed.

Animals↗
PubMed1995

Differential effects of glucocorticoids on insulin-like growth factor I action in cultured human fibroblasts.

Glucocorticoids act synergistically with insulin-like growth factor I (IGF-I) to stimulate DNA synthesis and replication of cultured human fibroblasts. In the present study, we further define glucocorticoid and IGF-I interactive effects on human fibroblast metabolism and growth. IGF-I stimulated dose-dependent increases in early metabolic events. Half-maximal effectiveness was seen at 5-8 ng/ml IGF-I, with mean maximal responses of 1.5-, 2-, and 6-fold for [3H]2-deoxyglucose uptake, [14C]glucose incorporation, and [14C]aminoisobutyric acid (AIB) uptake, respectively. A 48-hour preincubation with 10(-7) M dexamethasone markedly enhanced both the sensitivity and maximal effectiveness of IGF-I stimulation of AIB uptake. In contrast, dexamethasone had no effect on IGF-I-stimulated glucose uptake and utilization. Maximum specific binding of [125I]IGF-I to fibroblast monolayers was identical in ethanol control and glucocorticoid-treated cells, with 50% displacement at approximately 5 ng/ml IGF-I. In addition to its synergism with IGF-I, preincubation with dexamethasone augmented insulin and epidermal growth factor (EGF) stimulation of [3H]thymidine incorporation; dexamethasone had no effect on platelet-derived growth factor or fibroblast growth factor action. Two-dimensional gel electrophoresis identified two specific glucocorticoid-induced proteins in human fibroblast cell extracts with molecular weights of 45K and 53K and pIs of 6.8 and 6.3, respectively. These data indicate that IGF-I receptor-mediated actions in human fibroblasts are differentially modulated by glucocorticoids. Glucocorticoids are synergistic with IGF-I in stimulating mitogenesis and amino acid uptake, without having any apparent effect on IGF-I-stimulating glucose metabolism. Glucocorticoid enhancement of growth factor bioactivity may involve modulation of a regulatory event in the mitogenic signaling pathway subsequent to cell surface receptor activation.

Aminoisobutyric Acids↗
PubMed1988

Genetic manipulation of non-conventional yeasts by conventional and non-conventional methods.

In recent years, yeasts other than those belonging to the species Saccharomyces cerevisiae and Schizosaccharomyces pombe have become increasingly important in industrial processes. Species such as Pichia stipitis, Hansenula polymorpha, Zygosaccharomyces rouxii, Saccharomyces exiguus, Torulaspora delbrueckii, Yarrowia lipolytica and others whose perfect stage is known, can be manipulated genetically by classical methods, but those belonging to the genera Candida (C. utilis, C. tropicalis, C. bombicola, C. zeylanoides, C. boidinii, etc.), Brettanomyces, Cryptococcus, Rhodotorula, and others of the different form genera, cannot be treated in this way. Some, such as Schwanniomyces and Debaryomyces spp., which have a perfect stage, are still difficult to manipulate by conventional means. Genetic manipulation of these yeasts can be approached from two points of view; the first involving improvement of strains by cross-breeding within one species, and the second, the introduction of desirable genes from unrelated species and even from plants or animals. Two techniques are available for construction of industrially-useful strains from these yeasts: protoplast fusion and transformation with chimaeric plasmids containing the gene(s) it is desired to introduce into the recipient strain. The methods for the latter procedure are well known but can be laborious and time-consuming, especially if it is desired to introduce genes from plant or animal sources for production of enzymes, hormones, vaccines and similar products. Protoplast fusion is a simple technique which can be utilized in most laboratories and used for construction of improved yeast strains for brewing, baking, ethanol production and wine-making, either by the fusion of desirable strains of the same species which do not sporulate, or by introduction of genes from non-Saccharomyces species. Methods for fusion of species from different genera and isolation of the desired hybrids have been improved considerably in recent years. We have developed a method for isolation of strains carrying the desired genes by fusing a non-Saccharomyces species with an auxotrophic strain of Saccharomyces cerevisiae and selecting hybrids having the desired characteristics on appropriate media, after which the genes are transferred to the industrial strain by rare-mating, repeated protoplast fusion, or classical mating as required. The advantages and limitations of the method are under investigation.

Genetic Engineering↗
PubMed1993

Aerobic metabolism of carbon reserves by the "obligate anaerobe" Desulfovibrio gigas.

The sulfate-reducing bacterium, Desulfovibrio gigas, is shown by in vivo 31P-NMR to be capable of generating NTP from the utilization of internal carbon reserves both in anaerobic and in aerobic conditions. Acetate, glycerol and ethanol are the major end-products, but the production of alcohols decreases strongly when oxygen is present. When the glycolytic pathway is inhibited with fluoride, NTP levels decrease drastically but can be remarkably restored when an electron acceptor, such as oxygen, is provided. Our data are in favour of a NADH-linked electron transfer chain enabling transfer of reducing power derived from polyglucose to oxygen which provides this so-called "strict anaerobe" with the capability of surviving to oxic environments.

Acetates↗
PubMed1986

Bacteroides xylanolyticus sp. nov., a xylanolytic bacterium from methane producing cattle manure.

As part of a study of the biogas production from cattle waste, xylanolytic bacteria were isolated from enrichments of fermenting cattle manure. From 34 isolates, mostly Gram-negative rods, a typical strain was investigated in more detail. It was an anaerobic non-sporeforming, Gram-negative rod, which was motile with peritrichous flagella. This organism fermented xylan and many soluble sugars (glucose, cellobiose, mannose, xylose, arabinose). Other hemicelluloses such as gum xanthan, laminaran, locust bean gum, and gum arabic were not utilized. It also could not use cellulose. Fermentation products were carbon dioxide, hydrogen, acetate and ethanol. The bacterium produced carboxymethylcellulase and xylanase, especially when growing on xylan. Growth was optimal between 25 degrees C and 40 degrees C and between pH 6.5 and 7.5. The guanine plus cytosine content of the DNA was 34.8 +/- 0.8%. The isolate was identified as a member of the genus Bacteroides, and a new species is proposed: Bacteroides xylanolyticus (xylan dissolving). The type strain of B. xylanolyticus is strain X5-1 (DSM 3808).

Animals↗
PubMed1994

Purification and molecular characterization of the NAD(+)-dependent acetaldehyde/alcohol dehydrogenase from Entamoeba histolytica.

A bifunctional 95 kDa polypeptide (EhADH2) harbouring acetaldehyde dehydrogenase and alcohol dehydrogenase activities was purified to homogeneity from trophozoite extracts of the protozoan parasite Entamoeba histolytica. Kinetic studies revealed that the enzyme utilizes NAD+ rather than NADP+ as cofactor. Km values for acetyl-CoA, acetaldehyde and ethanol were found to be 0.015, 0.15 and 80 mM respectively in the presence of 0.2 mM NAD+. The primary structure of EhADH2 as deduced from respective amoebic DNA sequences showed striking similarity to the trifunctional AdhE protein of Escherichia coli and the bifunctional AAD protein of Clostridium acetobutylicum. Alignment with a number of aldehyde dehydrogenases and alcohol dehydrogenases from various species suggested that the two catalytic functions of EhADH2 are located on separate parts of the molecule. By cross-linking experiments and electron-microscopic analysis, native EhADH2 was found to be organized in a homopolymeric fashion consisting of more than 20 associated promoters which form rods about 50-120 nm in length.

Alcohol Dehydrogenase↗
PubMed1983

Membrane properties of isolated winter wheat cells in relation to icing stress.

Isolated cell preparations of winter wheat (Triticum aestivum L.) were utilized to examine the effect of ice encasement at -1 degrees C and exposure to ethanol on metabolic and biochemical properties of cells. Following icing and ethanol treatments, passive efflux of amino acids increased gradually with duration of exposure to the stress, and closely paralleled the decline in viability of cells. In contrast, uptake of (86)Rb declined much more rapidly than viability following exposure to icing or ethanol. Electron spin resonance spectroscopy studies revealed no significant change in molecular ordering within the cell membranes following icing or exposure to ethanol, whereas a small but significant increase in order was detected in the noniced controls. O(2) consumption by isolated cells declined only gradually due to icing and ethanol treatments, and remained relatively high even when cell viability was severely reduced. These results indicate that the plasma membrane is a primary site of injury during ice encasement and that damage to the ion transport system is the earliest manifestation of this injury.

Journal Article↗