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PubMedDate not supplied

Efficient intracellular delivery of oligonucleotides formulated in folate receptor-targeted lipid vesicles.

In this study, a novel lipid vector has been developed for targeted delivery of oligodeoxynucleotides (ODN) to tumors that overexpress folate receptor. This is based on a method developed by Semple et al. (1), which utilizes an ionizable aminolipid (1,2-dioleoyl-3-(dimethylammonio)propane, DODAP) and an ethanol-containing buffer system for encapsulating large quantities of polyanionic ODN in lipid vesicles. Folate is incorporated into the lipid vesicles via a distearoylphosphatidylethanolamine-poly(ethylene glycol) (DSPE-PEG) spacer. These vesicles are around 100-200 nm in diameter with an ODN entrapment efficiency of 60-80%. Folate mediated efficient delivery of ODN to KB cells that overexpress folate receptor. Uptake of folate-targeted lipidic ODN by KB cells is about 8-10-fold more efficient than that of lipidic ODN without a ligand or free ODN. This formulation is resistant to serum. Thus, targeted delivery of ODN via this novel lipid vector may have potential in treating tumors that overexpress folate receptors.

Base Sequence↗
PubMed1997

Sporomusa silvacetica sp, nov., an acetogenic bacterium isolated from aggregated forest soil.

Sporomusa silvacetica sp. nov. DG-1T (= DSMZ 10669T) (T = type strain) was isolated from well-drained, aggregated forest soil (pH 6.0) in east-central Germany. The cells were obligately anaerobic, slightly curved rods and were motile by means of laterally inserted flagella on the concave side of each cell. Typical cells were approximately 3.5 by 0.7 micron. Cells stained weakly gram positive, but thin sections revealed a complex multilayer cell wall. Spores were spherical and distended the sporangia. Growth and substrate utilization occurred with ferulate, vanillate, fructose, betaine, fumarate, 2,3-butanediol, pyruvate, lactate, glycerol, ethanol, methanol, formate, and H2-CO2. With most substrates, acetate was the primary reduced end product and was produced in stoichiometries indicative of an acetyl-coenzyme A pathway-dependent metabolism. Fumarate was dismutated to succinate and acetate. Methoxyl and acrylate groups of various aromatic compounds were O-demethylated and reduced, respectively. Yeast extract was not required for growth. Cells grew optimally at approximately 30 degrees C and pH 6.8; under these conditions and with fructose as the substrate, the doubling time was approximately 14 h. The lowest temperature that supported growth was between 5 and 10 degrees C. The carbon monoxide dehydrogenase and hydrogenase activities were approximately 9 and 102 mumol min-1 mg of protein-1, respectively. A type b cytochrome was detected in the membrane. The G + C content was approximately 43 mol%. Phylogenetic analysis of the 16S ribosomal DNA indicated that DG-1T was most closely related to members of the genus Sporomusa in the Clostridium subphylum of the gram-positive bacteria.

Base Composition↗
PubMed1996

Effect of CO2 on the fermentation capacities of the acetogen Peptostreptococcus productus U-1.

The fermentative capacities of the acetogenic bacterium Peptostreptococcus productus U-1 (ATCC 35244) were examined. Although acetate was formed from all the substrates tested, additional products were produced in response to CO2 limitation. Under CO2-limited conditions, fructose-dependent growth yielded high levels of lactate as a reduced end product; lactate was also produced under CO2-enriched conditions when fructose concentrations were elevated. In the absence of supplemental CO2, xylose-dependent growth yielded lactate and succinate as major reduced end products. Although supplemental CO2 and acetogenesis stimulated cell yields on fructose, xylose-dependent cell yields were decreased in response to CO2 and acetogenesis. In contrast, glycerol-dependent growth yielded high levels of ethanol in the absence of supplemental CO2, and pyruvate was subject to only acetogenic utilization independent of CO2. CO2 pulsing during the growth of CO2-limited fructose cultures stopped lactate synthesis immediately, indicating that CO2-limited cells were nonetheless metabolically poised to respond quickly to exogenous CO2. Resting cells that were cultivated at the expense of fructose without supplemental CO2 readily consumed fructose in the absence of exogenous CO2 and formed only lactate. Although the specific activity of lactate dehydrogenase was not appreciably influenced by supplemental C02 during cultivation, cells cultivated on fructose under CO2-enriched conditions displayed minimal capacities to consume fructose in the absence of exogenous CO2. These results demonstrate that the utilization of alternative fermentations for the conservation of energy and growth of P. productus U-1 is augmented by the relative availability of CO2 and growth substrate.

Carbon Dioxide↗
PubMedDate not supplied

[Anaerobic cellulolytic fungi from cattle rumen].

Two anaerobic fungal strains, Neocallimastix sp. 71 and Piromonas sp. 12, were isolated from cattle rumen. Their motile zoospores have either one (strain 12) or several (strain 71) flagella. The spores get attached to a substrate, germinate and form a branched rhizoid with one sporangium within which a new generation of zoospores appears after 24 h, on the average. Both fungi have a temperature optimum at 39 degrees C and a pH optimum at 6.5-6.7. They can utilize a wide spectrum of mono-, oligo- and polysaccharides. Strain 71 produces acetate, formate, ethanol and lactate while strain 12 forms acetate, formate, lactate and traces of ethanol as main products accumulated upon the fermentation of wheat straw, filter paper and cellobiose in a liquid semidefined medium. Both strains exert the activities of CMCase, Avicelase, cellobiase, amylase and xylanase.

Anaerobiosis↗
PubMed1989

[A sarcoma-static new species of Pseudomonas, Pseudomonas jinanensis sp. nov].

A strain of Gram negative bacteria was isolated from the surface soil of Wuying Hill at Jinan, Shandong province with Gause's medium in 1973. It is a strain of antagonistic bacteria for hysterocervicoma, hepatoma and melanoma of mice screened from 2100 strains of bacteria. It is also antagonistic to Staphylococcus aureus, Bacillus subtilis and Micrococcus. It is a Gram negative bacterium with lophotrichous polar flagella. Straight rods in shape or with a little slightly curved rods, 0.5-0.6 X 1-2 microns, randomly arranged, poly-beta-hydroxybutyrate granules are accumulated in cells after 2-5 days cultivation. Water green soluble pigment and green fluorescent pigment are produced. Respiratory metabolism, chemoorganotroph, many carbon-containing organic compounds can be used as carbon sources, such as glucose, trehalose, ethanol, cellulobiose, fucose, arginine and betaine, but propionic acid or tartaric acid is not utilized. Inorganic nitrogen containing compounds can be used ae the sole source of nitrogen. No growth factor is necessary for growth. Gelatin is hydrolyzed. Starch and cellulose are not hydrolyzed. Nitrate is not reduced. Arginine dihydrolase is produced. Levan is produced from sucrose. Growth occurs from 7 degrees C to 37 degrees C and from pH 5.65-8.40. No growth occurs at 40 degrees C and at pH value below 4.86. It can not grow autotrophically with hydrogen. Its G + C contents in DNA is 58.1 mol%. DNA-DNA hybridization experiments reveals a relatedness value of 58.6% between this strain and Ps. fluorescens. The above evidence shows that this strain differs from all species known in Pseudomonas, such as Pseudomonas fluorescens group. Pseudomonas caryophylli, Pseudomonas cepacia, Pseudomonas marginata, Pseudomonas acidovorans, Pseudomonas testosteroni and Pseudomonas delafieldii.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Tissue distribution and hepatic ultrastructural effects of aflatoxin B1 in Japanese quail.

Fertile, Japanese quail eggs were injected with graded levels of aflatoxin B1 (AFB1) dissolved in ethanol. Hatchability varied inversely with the quantity of AFB1 administered. The generated quail were utilized in studies of AFB1 distribution and its effects upon hepatic ultrastructure. Putative AFB1 was detected within the livers of two quail 6 h after dosing, although the AFB1 distribution was inconsistent between the individuals. Electron microscopy revealed that liver parenchymal cells from AFB1-treated exhibited mitochondrial pleomorphism 6 h after dosing, a rare observation in control tissue.

Aflatoxin B1↗
PubMedDate not supplied

[A new psychroactive bacterium, Clostridium fimentarium, isolated from cattle manure fermented at a low temperature].

A new psychroactive saccharolytic anaerobic organism was isolated from cattle manure digested at 6 degrees C. The organism is a Gram-positive oligosporous motile rod measuring 0.5-0.6 by 2.1-5.0 microns. It utilizes glucose, fructose, maltose, arabinose, xylose, cellobiose, galactose, and mannose. Fermentation products are acetate, ethanol, lactate, formate, H2, and CO2. Growth is possible within a temperature range of 1-30 degrees C, with an optimum at 20-25 degrees C, and a pH range of 5.5-8.3, with an optimum at 6.8. The DNA GC content is 35.6 mol %. The organism was classified as a new species of the genus Clostridium, C. fimetarium sp. nov. The type strain is Z-2189.

Animals↗
PubMed1993

Structural characterization of monellin in the alcohol-denatured state by NMR: evidence for beta-sheet to alpha-helix conversion.

Two-dimensional 1H NMR spectroscopy and hydrogen exchange methods have been used to characterize the alcohol-denatured state of monellin. Monellin is a sweet tasting protein composed of two chains. In the native state, the A-chain consists entirely of beta-structure, and the B-chain contains both alpha- and beta-structure. Upon addition of either 50% ethanol or 50% trifluoroethanol (TFE), the native structure of monellin is disrupted resulting in an alcohol-denatured state with properties different from those of the random coil state. In the alcohol-denatured state, the far-UV circular dichroism (CD) spectrum displays a higher helical content relative to the native state and the intensity of the near-UV CD signal is completely lost. One-dimensional NMR studies show that there are approximately 14 amide protons protected from exchange with solvent in the alcohol-denatured state and that large portions of the protein exchange at a rate that is comparable to the exchange rate of the protein in urea. Utilizing hydrogen exchange trapping techniques, the slowly exchanging residues are identified at pH 2.0 in 50% ethanol and 50% TFE (A10-A15, A18, A19, A21, A24, and A39) and are found to be clustered on one region of the A-chain. Preliminary 2D NMR assignments show that in the alcohol-denatured state the A-chain of monellin undergoes structural reorganization, with one strand of the native state beta-sheet on the A-chain (residues A17-A30) becoming an alpha-helix in the alcohol-denatured state. The secondary structure of the A-chain in the alcohol-denatured state is different from the native state structure, although the slowly exchanging residues are similar.

Amino Acid Sequence↗
PubMed1984

[Mechanisms of high hemoglobin A1 in alcohol drinkers].

Hemoglobin A1 (HbA1) levels were significantly higher in healthy alcohol drinkers (HbA1 = 7.50%, n = 11) than in normal non-drinkers (HbA1 = 6.62%, n = 13). Ethanol was not able to change HbA1 level when ethanol was added to human whole blood in vitro. Acetaldehyde (AcCHO), although, markedly increased it. Glucose utilization in erythrocytes was stimulated by AcCHO. While it was completely blocked by sodium fluoride in the presence of AcCHO in the incubation medium, but sodium fluoride did not affect the formation of HbA1. AcCHO formed HbA1 with human purified hemoglobin in vitro. The level of HbA1 formed by AcCHO was significantly low when purified human hemoglobin used as a substrate in comparison with the use of whole blood. AcCHO and dihydroxyacetone phosphate reacted in the presence of aldolase. The reacted product, 5-deoxy-D-xylulose-1-phosphate, increased HhA1 level of human purified hemoglobin. It is suggested, the high level of HbA1 in healthy drinkers was caused by AcCHO, the first metabolite of ethanol. AcCHO formed addicts with human hemoglobin directly, and there might be other mechanisms of HbA1 formation due to AcCHO, such as 5-deoxy-D-xylulose-1-phosphate, which is the reacted product of AcCHO.

Acetaldehyde↗
PubMed1980

[Intravenous ethyl alcohol in metabolic resuscitation].

Intravenously administered ethyl alcohol may be effective as analgesic and hypotensive peripheric vasoactive drug. In the Intensive Care Departments parenteral ethanol administration is infrequent because no "sure dosage" can be suggested in adults and children. Liver, kidney and C.N.S. diseases can worsen; foetopathy can follow. Drug-ethanol interaction may be particularly important for some patients admitted in Intensive Care Departments. Often the potential caloric support cannot be fully utilized ("empty" calories) and seldom hyperventilation, hyperlactacidemia and impaired protein synthesis can follow.

Critical Care↗
PubMed2003

Bioconversions of maize residues to value-added coproducts using yeast-like fungi.

Agricultural residues are abundant potential feedstocks for bioconversions to industrial fuels and chemicals. Every bushel of maize (approximately 25 kg) processed for sweeteners, oil, or ethanol generates nearly 7 kg of protein- and fiber-rich residues. Currently these materials are sold for very low returns as animal feed ingredients. Yeast-like fungi are promising biocatalysts for conversions of agricultural residues. Although corn fiber (pericarp) arabinoxylan is resistant to digestion by commercially available enzymes, a crude mixture of enzymes from the yeast-like fungus Aureobasidium partially saccharifies corn fiber without chemical pretreatment. Sugars derived from corn fiber can be converted to ethanol or other valuable products using a variety of naturally occurring or recombinant yeasts. Examples are presented of Pichia guilliermondii strains for the conversion of corn fiber hydrolysates to the alternative sweetener xylitol. Corn-based fuel ethanol production also generates enormous volumes of low-value stillage residues. These nutritionally rich materials are prospective substrates for numerous yeast fermentations. Strains of Aureobasidium and the red yeast Phaffia rhodozyma utilize stillage residues for production of the polysaccharide pullulan and the carotenoid astaxanthin, respectively.

Ethanol↗
PubMedDate not supplied

ERP responses to ethanol and diazepam administration in squirrel monkeys.

The effects of oral administration of two doses of ethanol (0.75 and 1.5 g/kg) and a single dose of diazepam (2.5 mg/kg) on electrophysiological responses to an event related potential (ERP) paradigm were investigated in the squirrel monkey Saimiri Sciureus. A passive auditory "oddball plus novel" tone sequence was utilized to elicit N1, P2, N2, and P3-like components which resembled waveforms obtained from human subjects under the same conditions. Ethanol at both doses was found to produce a significant decrease in the amplitude of the N1 component. The higher ethanol dose also produced a significant decrease in the amplitude of the P3-like component. Ethanol at both doses was observed to produce significant increases in the variability of the amplitude of these components as measured from trial to trial. Diazepam, like ethanol, also induced a significant decrease in the N1 component of the ERP but did not affect the later waves. These results suggest that ERPs can reliably assess the electrophysiological actions of ethanol in monkeys.

Animals↗
PubMed1990

Beneficial effect of 1,3-butanediol on cerebral energy metabolism and edema following brain embolization in rats.

We assessed the effect of 1,3-butanediol on cerebral energy metabolism and edema after inducing multifocal brain infarcts in 108 rats by the intracarotid injection of 50-microns carbonized microspheres. An ethanol dimer that induces systemic ketosis, 25 mmol/kg i.p. butanediol was injected every 3 hours to produce a sustained increase in the plasma level of beta-hydroxybutyrate. Treatment significantly attenuated ischemia-induced metabolic changes by increasing the concentrations of phosphocreatine, adenosine triphosphate, and glycogen and by reducing the concentrations of pyruvate and lactate. Lactate concentration 2, 6, and 12 hours after embolization decreased by 13%, 44%, and 46%, respectively. Brain water content increased from 78.63% in six unembolized rats to 80.93% in 12 saline-treated and 79.57% in seven butanediol-treated rats 12 hours after embolization. (p less than 0.05). The decrease in water content was associated with significant decreases in the concentrations of sodium and chloride. The antiedema effect of butanediol could not be explained by an osmotic mechanism since equimolar doses of urea or ethanol were ineffective. Our results support the hypothesis that the beneficial effect of butanediol is mediated through cerebral utilization of ketone bodies arising from butanediol metabolism, reducing the rate of glycolysis and the deleterious accumulation of lactic acid during ischemia.

3-Hydroxybutyric Acid↗
PubMed1999

Effects of escins Ia, Ib, IIa, and IIb from horse chestnuts on gastric emptying in mice.

Inhibitory effects of the saponin fraction and its principal constituents, escins Ia, Ib, IIa, and IIb, from horse chestnuts on gastric emptying were investigated in mice loaded with a non-nutrient or nutrient meal. The saponin fraction and escins Ia-IIb inhibited gastric emptying of a 1.5% carboxymethyl cellulose sodium salt (CMC-Na) meal by 11.1-54.2% (12.5-200 mg/kg). Escins Ia-IIb (50 mg/kg) also inhibited gastric emptying of a 40% glucose meal by 21.1-23.5% except for escin Ia, a milk meal by 18.4-33.1%, and a 30% ethanol meal by 13.5-15.9%. The effects of escins Ia-IIb on gastric emptying of the CMC-Na meal were attenuated by pretreatment with streptozotocin (100 mg/kg, i.v.), capsaicin (75 mg/kg in total, s.c.), or insulin (1 U/kg, s.c.). The effect of insulin was reduced by glucose (2 g/kg, i.v.) which can directly nourish the brain, but not by fructose (2 g/kg, i.v.) which cannot be utilized by the brain. The effects of escins Ia-IIb (50 mg/kg) were overridden in 60% ethanol-loaded mice, in which the central nervous system was suppressed by ethanol. These results suggest that capsaicin-sensitive sensory nerves and central nervous system partly participate in the effects of escins Ia-IIb.

Animals↗
PubMed1983

Na-Dithionite Promotes Photosynthetic Sulfide Utilization by the Cyanobacterium Oscillatoria limnetica.

The light- and sulfide-dependent induction process leading to photosystem I-mediated sulfide utilization by Oscillatoria limnetica, for either H(2) evolution or CO(2) photoassimilation, was studied. The identical dependence on pH of the lag length, the inhibition of leucine incorporation and final H(2)S concentration imply that the latter exerts a deleterious effect on nonadapted cells.Na-dithionite (Na(2)S(2)O(4)), Na-sulfite (Na(2)SO(3)), or ethanol cannot serve as photosynthetic electron donors. However, when these compounds were added to the sulfide-containing system, the need for induction was eliminated. At pH 6.9, in the presence of 3.5 millimolar sulfide, these substances (at concentrations of 10 millimolar, 5 millimolar, or 0.4 molar, respectively) completely abolished the delay preceding sulfide-dependent H(2) evolution. It is suggested that all three compounds expose a site capable of directly accepting sulfide electrons.Only dithionite could adapt the cells to sulfide utilization on its own. Sulfite or ethanol acted only in the presence of sulfide. It is implied that this specific activity of dithionite is related to its characteristic low redox potential.Sulfide-dependent H(2) evolution was insensitive to 3-(3,4-dichlorophenyl)-1,1-dimethylurea, but was inhibited by the plastoquinone antagonist 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, in the presence as well as in the absence of dithionite. In both cases, therefore, the plastoquinone was implied in the electron transport from sulfide.

Journal Article↗
PubMed1992

The pharmacological differences and similarities between stress- and ethanol-induced gastric mucosal damage.

Stress- and ethanol-induced gastric mucosal damage are the two commonly used ulcer models in animals. They share some of the similarities but also have differences in the etiology of gastric ulceration. This article reviews the influences of various protective drugs on these two types of gastric damage in rats. Verapamil (a calcium antagonist) or N-ethylmaleimide (a sulfhydryl depletor) prevents cold restraint-, but potentiates ethanol-provoked gastric lesion formation. N-Acetylcysteine (a mucolytic agent) and acetaminophen (an antipyretic analgesic) have the opposite actions. Prostaglandins provide a much better antiulcer effect on ethanol-induced lesions. Cimetidine (a histamine H2-receptor antagonist) prevents only stress-induced mucosal damage. These differences in drug actions indicate that stress and ethanol may have dissimilar ulcerogenic mechanisms in rats. On the other hand, carbenoxolone (a mucus inducer), histamine H1-receptor antagonists, leukotriene inhibitors (FPL 55712 and nordihydroguaiaretic acid) and mast cell stabilizers (like zinc compounds, sodium cromoglycate, FPL 52694 and ketotifen), all protect against gastric mucosal damage by stress or ethanol in rats. However, the role of gastric sulfhydryls in both types of gastric lesions is still controversial. These findings imply that the two types of lesion formation share some of the ulcerogenic mechanisms. This communication attempts to analyze the various findings and to relate them to the etiology of stress and ethanol-induced gastric lesions. It also summarizes the uses, and the antiulcer mechanisms, of the drugs that have been studied utilizing these two animal ulcer models, and suggests their possible implications in man.

Acetaminophen↗
PubMed2001

The mitochondrial consequences of uncoupling intact cells depend on the nature of the exogenous substrate.

In isolated mitochondria the consequences of oxidative phosphorylation uncoupling are well defined, whereas in intact cells various effects have been described. Uncoupling liver cells with 2,4-dinitrophenol (DNP) in the presence of dihydroxyacetone (DHA) and ethanol results in a marked decrease in mitochondrial transmembrane electrical potential (DeltaPsi), ATP/ADP ratios and gluconeogenesis (as an ATP-utilizing process), whereas the increased oxidation rate is limited and transient. Conversely, when DHA is associated with octanoate or proline, DNP addition results in a very large and sustained increase in oxidation rate, whereas the decreases in DeltaPsi, ATP/ADP ratios and gluconeogenesis are significantly less when compared with DHA and ethanol. Hence significant energy wastage (high oxidation rate) by uncoupling is achieved only with substrates that are directly oxidized in the mitochondrial matrix. Conversely in the presence of substrates that are first oxidized in the cytosol, uncoupling results in a profound decrease in mitochondrial DeltaPsi and ATP synthesis, whereas energy wastage is very limited.

Adenosine Diphosphate↗
PubMedDate not supplied

Effects of nicotine and nicotine/ethanol on human placental amino acids transfer.

Ethanol abuse and smoking during pregnancy both result in decreased offspring weight. One explanation for this may be impaired placental nutrient transport. This study assessed this possibility utilizing the 4-hr perfused human placental system and human placental vesicles exposed to "physiological," 0.2 microM and large (about 20 microM) nicotine concentrations alone, as well as nicotine combined with ethanol, 200 or 400 mg/ml, for up to 48 hr. Two nonmetabolizable amino acids, alpha-aminoisobutyric acid (AIB) and cycloleucine (CLEU) were used as probes. Nicotine was measured by gas chromatography in the placental perfusion system and vesicles and verified as to concentration. There was no statistically significant evidence of decreased transport of these amino acids with exposure to nicotine alone or nicotine and ethanol together in either test system. Thus, brief exposure to nicotine and ethanol does not impair amino acid transport by the human placenta.

Amino Acids↗