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Absorption and elution of anti-DLA-A and B antibodies with acetone-dried dog spleen powder.

Acetone-dried powders were prepared from the spleens of DLA-serotyped dogs and assayed for their specific absorptive properties for anti-DLA-A and B antisera. The specific activity of two different antisera (anti-DLA-A9 and anti-DLA-B13) was readily absorbed with acetone-dried powders prepared from the spleens of dogs of the corresponding DLA types. This specific activity was recovered by the elution of the powder used for the serum absorption. The same method was used to narrow the specificity of a highly polyspecific antiserum. The comparison of the absorption properties of acetone-dried spleen powder versus fresh spleen cells shows that the treatment with acetone does not modify qualitatively and quantitatively the DLA specificities.

Acetone↗

Conservation of active ribosomes in acetone-treated cells of Tetrahymena pyriformis.

The preparation of an acetone powder of cells of Tetrahymena pyriformis GL is described. A comparison of ribosomal particles isolated from acetone-treated and untreated cells shows that structurally and functionally intact ribosomes can be isolated from acetone-treated cells. Fully active ribosomes have been isolated from acetone powder of Tetrahymena that had been stored for more than 6 months at 4 degrees C. Thus, this procedure allows easy storage of large quantities of cells for the bulk preparation of active ribosomes.

Acetone↗

The relationships between the fertility of dairy cows and clinical and biochemical measurements, with special reference to plasma glucose and milk acetone.

Blood and milk samples were taken at first insemination in 352 dairy cows from 18 herds for charting the relation between clinico-chemical parameters and fertility rate. Neither total protein, albumin, globulin, AST, bilirubin, bile acids, calcium, magnesium and phosphorus in blood nor urea in milk was significantly related to the rate of pregnancy. On the other hand, there was a significant difference for plasma glucose between cows that became pregnant and those that did not. Cows with low plasma glucose concentrations at first insemination also had low values four and seven weeks after calving, indicating that it is primarily cows with chronically low blood glucose which are likely to have reduced fertility. At first insemination there was no increase in the concentration of acetone in milk of cows with reduced fertility. However, they had had increased milk acetone concentrations three to five weeks after calving. It therefore seems possible to evaluate the risk of reduced fertility by measuring either blood glucose or milk acetone some weeks after calving. Owing to the difficulties associated with the sampling and laboratory techniques for glucose, analyses of milk acetone are more suitable under field conditions.

Acetone↗

Effect of in vivo chromate, acetone and combined treatment on rat liver in vitro microsomal chromium(VI) reductive activity and on cytochrome P450 expression.

Cytochrome P450 (P450IIE1) in rat has previously been shown to exhibit high chromate [Cr(VI)] reductase activity (Mikalsen et al. 1991). The present study reports on the effect of chromate treatment in vivo in rats and on the modulating effect of acetone + fasting on chromium(VI) toxicity. No effect of intraperitoneal injection with 5 mg chromate/kg was observed, whereas 15 mg chromate/kg decreased the liver microsomal Cr(VI) reductase activity by about 30% in in vitro microsomal incubations. In addition, the P450 and cytochrome b5 contents were decreased by about 30% and 25% respectively. Acetone + fasting caused increases of total microsomal P450 and cytochrome b5 contents, associated with similar increases in apoproteins P450IIE1 and P450IIB1 + 2, and their corresponding mRNA, and apoprotein NADPH-P450 reductase, as well as NADPH-P450 reductase and microsomal Cr(VI) reductive activities. Related to acetone + fasting alone, when given in combination with chromate (15 mg/kg) the Cr(VI) reductive activity was decreased by about 30%, associated with decreases in the P450 and cytochrome b5 contents, 65% and 35% respectively. This further reduced the apoprotein levels of P450IIB1 + 2, P450IIE1, and NADPH-P450 reductase to 90%, 60%, and 40%, respectively, and the mRNA levels of P450IIB2 and P450IIE1. No effect was observed on NADPH-P450 reductase activity. This dose also caused some macroscopic alterations in the liver. In contrast, the P450IIE1 apoprotein level in the lung was apparently stabilized or even increased by chromate in rats treated with acetone + fasting.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetone↗

Comparative efficacy and toxicity of a ribosomal vaccine, acetone-killed cells, lipopolysaccharide, and a live cell vaccine prepared from Salmonella typhhimurium.

The protective and toxic properties of a ribosomal vaccine prepared from Salmonella typhimurium W118-2 were systematicaly compared with those of an acetone-killed whole cell vaccine, purified lipopolysaccharide, and living cells in CD-1 mice. Tests of graded immunizing doses of each vaccine against several challenge doses of live strain W118-2 showed that, although the protection given by ribosomes approached the levels of protection conferred by living organisms, acetone-killed cells administered in appropriate dosages provided levels of protection comparable to that of ribosomes. Lipopolysaccharide was found to be significantly less protective than the other vaccines. On a dry-weight basis, ribosomes were the least toxic with a 50% toxic dose (TD50) of 5,000 microgram; acetone-killed cells had an intermediate TD50 of 1,400 microgram; and lipolysaccharide was the most toxic, with a TD50 of 320 microgram. The dose of each vaccine that protected 50% of the mice against a challenge of 1,00 times the 50% lethal dose was determined and divided by the TD50 to give the therapeutic index. This ratio also indicated that the ribosomes and acetone-killed cells were equally effective, whereas lipopolysaccharide was markedly inferior.

Acetone↗

Skin prick tests with solutions of acid anhydrides in acetone.

Various low molecular acid anhydrides can act as haptens and induce type I allergies. Immunoglobulin E can be detected by radioallergosorbent test (RAST) with the protein conjugates of the respective anhydrides, which are commercially available for some substances. Being almost nonsoluble in water, these species cannot be applied in a skin prick test as a water solution. We will introduce a practicable skin prick test using acetonic solutions of the haptens. 136 persons were tested with acetone for negative control and histamine for positive control. Tests were carried out with 1 and 5% acetonic solutions of phthalic anhydride (PA). The RASTs with conjugates of the PA were performed with 111 subjects who had been exposed to acid anhydrides and with 5 people who possibly had been exposed. In 14 cases, when immediate-type allergy was suspected clinically, a test was performed in addition to the respective anhydride used at the workplace. In order to avoid unspecific histamine liberation, we tested 20 volunteer control persons who had not been occupationally exposed to acid anhydrides. In the prick test performed with acetonic PA solutions, 10 of the occupationally exposed subjects, who had also had a positive result in the RAST with protein conjugates of the PA, showed positive skin reactions, and in 2 further cases, in which the skin reaction was positive, the results of the RAST were just below the limit of a positive evaluation. The skin reaction was positive in 4 cases, in spite of a negative RAST result, while the prick test was reported to be negative in 3 cases although the RAST results were considered to be positive.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetone↗

Evidence for the existence of an acetone soluble renin inhibiting factor in normal human plasma.

After addition of exogenous human renin, the in vitro rate of angiotensin I generation is faster in plasma of patients with chronic renal insufficiency and, to a lesser extent, in plasma of patients with essential hypertension than in plasma from normotensive control subjects. The increased reactivity of renin in hypertensive and uremic plasma is not related to differences of endogenous renin activity, angiotensinase activity, renin substrate concentration, or substrate reactivity. Addition of normal, hypertensive, and uremic plasma to a human renin-sheep renin substrate system inhibited the rate of angiotensin generation, although significantly less inhibition was observed with uremic plasma. The reactivity of renin increased in normal plasma but not in uremic plasma after treatment with 95% acetone. After acetone extraction renin reactivity in normal and plasma inhibited the rate of angiotensin generation in a renin-renin substrate system. Less inhibition occurred with the acetone extract from a pool of uremic plasma. These results provide evidence for the existence of a naturally occurring acetone soluble renin inhibiting factor in normal and uremic plasma. The increased reactivity of renin in uremic plasma may be related to a deficiency of this factor.

Acetone↗

Recovery of prolactin from acetone-dried human pituitary glands.

An improved method is described for the purification of prolactin from the same batch of acetone-dried pituitary glands which is used for the isolation of other anterior pituitary hormones. The residue obtained after the extraction of the glycoprotein fraction from the pituitary acetone powder was extracted for growth hormone. The supernatant after the precipitation of growth hormone at 50% ethanol concentration was adjusted to 85% in ethanol. To the supernatant obtained from that step an equal volume of acetone was added to precipitate the 'PRL fraction.' This fraction was further purified by gel filtration on Sephadex G-100, ion-exchange chromatography on DEAE-cellulose and isoelectric focussing. A yield of 23 mg prolactin, 37 U/mg, was obtained per 1000 g pituitary acetone powder.

Acetone↗

Enhancement of fibrinolytic activity of human plasma in the presence of acetone.

The fibrinolytic system was studied in normal human plasma containing increasing concentrations of acetone up to 23.4 mmol l-1. Fibrinolytic activity measured as euglobulin clot lysis time [ECLT] and amidase activities toward chromogenic peptide substrates H-D-Valyl-L-Leucyl-L-Lysine-p-nitroanilide 2 HCl [S-2251], designed for plasmin determination, H-D-Valyl-L-Phenylalanyl-L-Lysine-p-nitroanilide 2 HCl [S-2390], designed for the determination of t-PA in plasma via plasminogen activation and H-D-Prolyl-L-Phenyl-Alanyl-L-Arginine-p-nitro-anilide 2 HCl [S-2302], designed for the determination of kallikrein and activated Hageman factor, increased when 15.7 mmol l-1 concentration of acetone was reached. A parallel increase of esterolytic [substrate: naphthol-AS-acetate] activity was observed in euglobulin fractions. Crossed immunoelectrophoresis [CIE] revealed changes in fibrinogen profiles of plasma enriched with acetone as compared to native plasma. These findings suggest that acetone present in plasma in concentrations comparable to those found in some pathological states might activate fibrinolytic system.

Acetone↗

Induction of rat hepatic mixed-function oxidases by acetone and other physiological ketones: their role in diabetes-induced changes in cytochrome P450 proteins.

1. To evaluate the role of ketone bodies in diabetes-induced changes in hepatic cytochrome P450 composition, rats were treated with acetone, 3-hydroxybutyrate or 1,3-butanediol. 2. Treatment with acetone enhanced the rat hepatic O-dealkylations of ethoxyresorufin and methoxyresorufin, and the hydroxylation of p-nitrophenol, but had no effect on lauric acid hydroxylation and ethylmorphine N-demethylation. Neither 3-hydroxybutyrate nor 1,3-butanediol modulated the metabolism of the above substrates. 3. Immunoblot analysis of hepatic microsomal proteins revealed that treatment with acetone increased the apoprotein levels of P4501A2, P4502B1/2 and P4502E1. 4. It is concluded that acetone is responsible, at least partly, for the diabetes-induced increase in hepatic microsomal P4501A2, P4502B1/2 and P4502E1 proteins but does not mediate the increases in the P4503A1 and P4504A1 proteins. On the basis of work from our own and other laboratories a mechanism for the diabetes-induced changes in hepatic cytochrome P450 proteins is proposed.

Acetone↗

Preparation of human serum albumin microspheres by a novel acetone-heat denaturation method.

Human serum albumin (HSA) microspheres have been produced in the size range of 200 nm to 10 microns by the controlled addition of acetone to an aqueous solution of HSA, followed by stabilization of the formed microspheres at an elevated temperature. Microspheres produced by this acetone-heat denaturation method could be stabilized at relatively low temperatures (75 degrees C) over a short time period (15-30 min). The acetone-heat denaturation method is different from the traditional oil/water technique for preparation of HSA microspheres, both in terms of production method and the avoidance of high temperatures (> 100 degrees C) and extended heating times (> 30 min) for stabilization. This paper describes the influence of process conditions, such as volumes of acetone and HSA concentration, on the formation of the microspheres and their morphological characteristics. A loading efficiency of 8 per cent was achieved for the HSA microspheres when using rose bengal as a model compound. The release of rose bengal from the microspheres in phosphate buffered saline at 37 degrees C was dependent on the presence of the surfactant Tween 80, and varied from a 5 per cent release in 14 days in the absence of surfactant to a 50 per cent release in 8 h in the presence of surfactant.

Acetone↗

[Comparison of the effect of acetone and isopropanol on lipid metabolism in rats].

Isopropanol and acetone administered to rats in conditions leading to a similar blood acetone level differ markedly in their effects on lipid metabolism. Isopropanol administration determines a fatty liver, which is mainly related to a defect in hepatic lipoprotein synthesis. Acetone administration gives only raise to a slight increase in the liver triacylglycerol level. It does not alter the [1-14C] palmitate, [1-14C] glycerol or [U-14C] leucine incorporation into blood lipoproteins. Acetone does thus not appear to play a preminent role in the isopropanol induced fatty liver which seems to be related mainly to a direct action of the alcohol itself.

1-Propanol↗

The adsorption of isopropanol and acetone by activated charcoal.

This study was performed to determine the adsorption of isopropanol and acetone by activated charcoal over a range of charcoal:solvent ratios. The charcoal binding of isopropanol was studied in both hydrochloric acid and water solutions, while acetone was analyzed in water. Gram ratios of charcoal to solvent ranged from 1:1 to 20:1. After the addition of charcoal the solution was agitated and centrifuged. The supernatant was then analyzed by gas chromatography. Each increment in charcoal dose increased the percent adsorption of both isopropanol and acetone. At the 20:1 ratio 87-92% of the solvent was bound by activated charcoal. In vivo study is needed to determine if activated charcoal therapy can shorten the half-life of isopropanol and acetone and decrease the duration of supportive care needed following ingestions of these solvents.

1-Propanol↗

Measurement of serum isopropanol and the acetone metabolite by proton nuclear magnetic resonance: application to pharmacokinetic evaluation in a simulated overdose model.

The purpose of this investigation was to 1) compare the performance of proton nuclear magnetic resonance spectroscopy to gas chromatography head-space analysis in the measurement of serum isopropanol and its metabolite, acetone, obtained during a simulated overdose, and 2) compare pharmacokinetic parameters obtained using the two analytical techniques. Three healthy volunteers ingested 0.6 mL/kg of 70% isopropanol and blood samples were obtained at baseline, 0.16, 0.33, 0.66, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12.0, and 24.0 hours post-ingestion. Resulting sera were analyzed by gas chromatography head-space analysis and proton nuclear magnetic resonance spectroscopy for determination of isopropanol and acetone concentrations. A correlation between concentrations quantitated by gas chromatography head-space analysis versus proton nuclear magnetic resonance spectroscopy was determined using linear regression. Pharmacokinetic disposition parameters were determined from serum concentration-time data and compared using analysis of variance. For isopropanol, the linear regression equation which describes the relationship between gas chromatography head-space analysis and proton nuclear magnetic resonance spectroscopy was y = 1.041x - 2.180 (r2 = 0.995, p < 0.0001); for acetone, y = 1.022x - 0.946 (r2 = 0.984, p < 0.0001). Pharmacokinetic disposition parameters derived from the two analytical methods were comparable. Proton nuclear magnetic resonance spectroscopy can be used to rapidly quantitate serum isopropanol and acetone concentrations in the same sample when gas chromatography head-space analysis is unavailable. Also, proton nuclear magnetic resonance spectroscopy can be used to follow serial serum concentrations during an ingestion for the purpose of pharmacokinetic analysis.

1-Propanol↗

Acetone-induced changes in the toxicokinetics of 2,5-hexanedione in rabbits.

Male rabbits were intravenously injected with 2,5-hexanedione or 2,5-hexanedione plus acetone. A toxicokinetic analysis showed that a two-compartment model satisfactorily described the kinetics of 2,5-hexanedione in rabbits. Simultaneous dosing with acetone altered the toxicokinetic model which best described the plasma concentration versus time data. The model-independent parameter body clearance, calculated according to the trapezoidal rule, showed a decrease in the body clearance of 2,5-hexanedione in rabbits simultaneously injected with acetone. The results suggest that toxicokinetic interference may partly explain the neurotoxic potentiation of that which occurs in 2,5-hexanedione-induced axonopathy as a response to simultaneous exposure to acetone.

Acetone↗

Loss of virus specific epitopes on JE virus 'E' glycoprotein by acetone treatment.

BACKGROUND & OBJECTIVES: Some Japanese encephalitis (JE) virus strains have been placed in group II based on the loss of reactivity against Hs (H = HI positive; s = JE virus specific) group of monoclonal antibodies (MAbs) in haemagglutination-inhibition (HI) test employing sucrose acetone (SA) extracted antigens. Also acetone-fixation of cells infected with some of the virus strains results in the loss of immunofluorescence (IF) against virus specific MAbs. The present study was undertaken to elucidate the effect of acetone on virus specific haemagglutination (HA) epitopes expressed on 'E' glycoprotein of group II strains of JE virus. METHODS: Porcine kidney (PS) cells were infected with JE virus strains (2 group I Indian strains, 5 group II strains and one neutralization-escape variant of 733913 group I strain). HI and complement fixation (CF) tests were carried out employing both polyethylene glycol (PEG) precipitated and SA extracted antigens of JE virus. RESULTS: Employing PEG precipitated antigens, Indian strain G9473 showed titres ranging from 1:40 to 1:160 against all the four virus specific HsMAbs and strain 641686 (1:160) with one of the four MAbs (Hs-1) by HI test whereas their SA extracted antigens did not react at all. In contrast, CF was positive employing both SA and PEG antigens in the presence of all four HsMAbs. The reactivity shown by PEG antigens in the HI test was confirmed by blocking the HA activity with the respective MAb. SA antigens, though negative in the HI test, were positive by the blocking assay. Interestingly, some of the non-HI MAbs which were negative against SA antigens, showed positive HI reaction with PEG antigens. Also, additional epitopes on Japanese (Yoken), Sri Lankan (691004) and two Indian (755468 and 641686) JE virus strains were detected either by blocking HA or surface IF. INTERPRETATION & CONCLUSIONS: It seems that the acetone treatment might result in HA property of the antigen which is no longer inhibited by an antibody in the HI test. The characterization of such labile and conformation-dependent epitopes is currently been undertaken to elucidate their role either in protection or immunopathogenesis of JE.

Acetone↗

Concentrations of blood and urine ethanol, acetaldehyde, acetate and acetone during experimental hangover in volunteers.

Blood and urine samples were analyzed for ethanol, acetaldehyde, acetate and acetone during experimental hangover in 6 healthy male volunteers (A, B, C, D, E, F). They drank freely for some 4 hr. In flushers (A, F) at 9 hr after ingestion (ethanol: 92 g, 1.2 g/kg and 1.3 g/kg), acetaldehyde levels were low in the blood, but high in the urine (37 microM, 45 microM). Heavy drinkers, non-flushers of B (ethanol: 176 g, 2.5 g/kg), C (157 g, 2.4 g/kg) and E (182 g, 2.9 g/kg) had a slightly high [lactate]/[pyruvate] ratio and 3-hydroxybutyrate in the blood at 11 hr after alcohol ingestion. Blood ethanol levels were dose dependent and blood acetaldehyde in B and C had a slightly high 6.3 microM and 8.0 microM 9 hr later, respectively. B, C and E had a high urine acetone concentration (100 microM over) in hangover. In C, in particular, urine acetate and acetone levels were unusually high. The ratio in blood (urine) among alcohol metabolites at 9 hr after drinking was approximately ethanol 1000 (1000): acetaldehyde 0.2-1.0 (0.1-5.9): acetate 36-163 (22-1554): acetone 1-11 (3-47).

Acetaldehyde↗

Effects induced in neutrophils by a precursor of triplet acetone.

The addition of a precursor of the enol form of isobutanal to neutrophils results in formation of triplet acetone, as attested to by emission from appropriate acceptors and cell damage (Nascimento et al., 1986 Biochim. Biophys. Acta 888, 337-342). The present study confirms the formation of triplet acetone by detection of the direct emission (lambda max 430 nm) and differentiates between effects produced by triplet acetone and by the enol substrate itself. Thus, triplet acetone: (1) enhances the release of ribonucleic acid; (2) promotes lipid peroxidation (N3(-)-inhibitable formation of thiobarbituric acid reactive products and concomitant light emission peaking at 480-500 nm); (3) increases myeloperoxidase activity, presumable as a result of damage and consequent increased exposure of the enzyme. On the other hand, the enol greatly enhances the release of protein(s) into the medium. These results confirm the utility of the neutrophil as a model system for the study of chemiexcitation processes induced at the cellular level. They also provide the first demonstration that an excited species formed at the cellular level may induce release of nucleic acids, thus reflecting the occurrence of deleterious processes in situ.

1-Propanol↗