Intracellular survival of viral and rickettsial agents in acetone at -60 C.
Viral and rickettsial agents, located intracellularly in cell monolayers, varied in their ability to survive exposure to acetone at -60 C.
Acetone↗
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Vaccination against Legionella pneumophila: serum antibody correlates with protection induced by heat-killed or acetone-killed cells against intraperitoneal but not aerosol infection in guinea pigs.
An aerosol model of Legionella infection has been established in guinea pigs. Infected animals showed growth of Legionella in their lungs, dissemination of organisms to the spleen, development of pneumonia and fever, and weight loss. Vaccination studies using heat-killed or acetone-killed cells were carried out, and guinea pigs were challenged intraperitoneally or by using the aerosol model of infection. Both vaccines were shown to give moderately high levels of protection against intraperitoneal challenge (28 to 145 50% lethal doses). Protection was found to be dose dependent and correlated with antibody levels as measured by enzyme-linked immunosorbent assay to an outer membrane antigen and by indirect immunofluorescence to heat-killed cells. In contrast, the same vaccination regimens that protected against intraperitoneal challenge failed to protect guinea pigs against aerosol challenge with comparable doses of Legionella, despite the presence of serum antibody. The results are discussed in terms of the possible requirements for immunity to aerosolized Legionella, including secretory immunoglobulin or cell-mediated immunity.
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The orphan response regulator HP1021 of Helicobacter pylori regulates transcription of a gene cluster presumably involved in acetone metabolism.
Helicobacter pylori is a gastric pathogen for which no nonhuman reservoir is known. In accordance with the tight adaptation to its unique habitat, the human stomach, H. pylori is endowed with a very restricted repertoire of regulatory proteins. Nevertheless, the three complete two-component systems of H. pylori were shown to be involved in the regulation of important virulence traits like motility and acid resistance and in the control of metal homeostasis. HP1021 is an orphan response regulator with an atypical receiver domain whose inactivation has a considerable impact on the growth of H. pylori. Here we report the identification of HP1021-regulated genes by whole-genome transcriptional profiling. We show that the transcription of the essential housekeeping genes nifS and nifU, which are required for the assembly of Fe-S clusters, is activated by HP1021. Furthermore, we demonstrate that the expression of a gene cluster comprising open reading frames hp0690 to hp0693 and hp0695 to hp0697 which is probably involved in acetone metabolism is strongly upregulated by HP1021. Evidence is provided for a direct regulation of the hp0695-to-hp0697 operon by the binding of HP1021 to its promoter region.
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Marking planes of surgical excision on breast biopsy specimens: use of artists' pigments suspended in acetone.
The performance of carbon and metallic inks, silver nitrate solution, and artists' pigments mounted in acetone was compared for marking the surface of surgical biopsy specimens. Using India ink is an unsatisfactory procedure because of slow drying, messiness, and spreading of the ink. It is concluded that use of artists' pigments has many advantages over other reagents, because of their rapid drying, resistance to tissue processing, and the ability to mark simultaneously many different planes of excision. Furthermore, the pigments are readily visible, are distinguishable from each other on microscopical examination, and the method entails little extra cost.
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Uptake, distribution, metabolism, and elimination of styrene in man. A comparison between single exposure and co-exposure with acetone.
Six male subjects were exposed for two hours during light physical exercise to 2.81 mmol/m3 (293 mg/m3) of styrene on one occasion and to a mixture of 2.89 mmol/m3 (301 mg/m3) of styrene and 21.3 mmol/m3 (1240 mg/m3) of acetone on another (combination study). About 68% of the dose (somewhat more than 4 mmol) of styrene was taken up. The arterial blood concentration of styrene reached a relatively stable level after about 75 minutes of exposure of about 18 and 20 mumol/l after the single and combined exposure, respectively. Calculated values of mean blood clearance were 1.9 l/min in the styrene study and 1.6 l/min in the combination study; the half life of styrene in blood was about 40 minutes in both studies. The concentration of non-conjugated styrene glycol increased linearly during exposure and reached about 3 mumol/l at the end of exposure and was eliminated with a half life of about 70 minutes. Styrene-7,8-oxide was detected and quantified in the blood in a complementary study. The half lives for the excretion of mandelic and phenylglyoxylic acid in the urine were about four and nine hours, respectively, in both studies.
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Labelled acetone and levulinic acid are formed when [14C]acetate is being converted to mycophenolic acid in Penicillium brevicompactum.
Evidence is presented that is compatible with the hypothesis that the farnesyl unit involved in the biosynthesis of mycophenolic acid in Penicillium brevicompactum is degraded by two successive oxidative cleavages at the double bonds distal from the phthalide nucleus. Acetone and levulinic acid are two non-aromatic degradation products.
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The extent to which resin can infiltrate dentin by acetone-based adhesives.
The combined methodologies of fractography and laser-Raman spectroscopic analysis were used for evaluation of the resin-dentin bonds made with wet and dry bonding. Resin-dentin-bonded beams were produced by means of 2 acetone-based adhesives (One-Step and Prime & Bond NT). The micro-tensile bond test was conducted, and the fractured surfaces of all specimens were examined by SEM and an image analyzer. The amount of resin infiltration within the hybrid layer was quantified by means of a laser-Raman spectroscope. In Raman analysis, the amount of resin impregnation within the hybrid layer of the dry bonding was found to be significantly lower (approximately 50%) than that in the wet one. Under fractographic analysis, a correlation was found between the bond strength and the failure mode. Based on those findings, it was suggested that the integrity between the bonding resin and the top of the hybrid layer played a major role in bond strength.
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Demonstration of species difference of placental alkaline phosphatase isozymes in acetone-fixed paraffin-embedded tissues.
The best preservation and discrimination of alkaline phosphatase isozymes by means of amino acid inhibition and heat treatment was obtained in sections of acetone-fixed paraffin-embedded tissues as compared to tissues fixed either in 95% ethanol or standard formalin. The results are illustrated in a study differentiating the isozymes of human and mouse placenta.
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Immunohistochemical demonstration of surface antigen of human lymphocytes with monoclonal antibody in acetone-fixed paraffin-embedded sections.
Although the majority of reported studies have used fresh-frozen sections in detecting surface antigen of lymphocytes in tissue via monoclonal antibody, detailed histological figures can not be obtained by this method. Nor can the antigenicity be preserved for any length of time. A new method for detecting the surface antigen of lymphocytes using fixed and embedded material is presented. Human spleens were fixed in cold acetone, embedded in low melting point paraffin wax, and the thin sections treated with hyaluronidase. Anti-T lymphocyte monoclonal antibody (anti-Leu-1, anti-Leu-2, anti-Leu-3) and anti-HLA-DR were applied on these sections, and the antigen was detected by the ABC (avidin-biotin-peroxidase complex) method. The results were then compared with those of fresh-frozen sections. There was no great difference in detecting T and B cells or their subsets, but the histological figures were substantially better preserved in sections prepared by the present method. Furthermore, the antigenicity was retained in the materials fixed and embedded for more than two years.
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Expression of 20 KD homologous restriction factor of complement on myocardial cells: an immunohistochemical study using acetone-fixed and paraffin-embedded tissues.
Expression of 20 KD homologous restriction factor of complement (HRF20) (CD59) in normal human heart was immunohistochemically examined with monoclonal antibody (MAb) 1F5. HRF20 was clearly demonstrated on the cell surface membrane and intercalated discs of myocardial cells throughout the ventricular walls. Therefore, this factor may protect normal cardiomyocytes from complement deposition, which has been demonstrated on infarcted cardiomyocytes. Expression of HRF20 was also observed on endothelial cells of the endocardium and on blood vessels, including arteries, capillaries, and veins, and on the Schwann-cell sheath of peripheral nerve fibers. In the present study, we found that the epitopes of HRF20 were well preserved for immunohistochemistry even in acetone-fixed and paraffin-embedded tissues, as well as in frozen tissues that have been conventionally used for HRF20. This method using paraffin sections allows for easier staining and greater accuracy in histological examination.
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Ab initio molecular orbital study of the reactivity of active alkyl groups. V. Nitrosation mechanism of acetone with syn-form of methyl nitrite.
The mechanisms of nitrosation of acetone through sodium enolate [CH3CO1CH2]-Na+ (1) or naked enolate [CH3CO1CH2]- (2) with methyl nitrite CH3O3NO2 (3), and the reactivity of the syn-form of 3 (syn-3) during the C-N bond formation process were investigated using ab initio molecular orbital (MO) methods. Our results have demonstrated the predominant formation of E-1-hydroxyimino-2-oxo-propane CH3COCH=NOH (4E) when the complex [CH3CO1CH2NO2(O3CH3)]-Na+ was produced kinetically via a metal-chelated pericyclic transition state (TS(CHELATED)), in which the O3 atom of syn-3 was coordinated to the Na+ atom of 1.
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[Sensitive screening of antifungal compounds from acetone extracts of medicinal plants with a Bio-Cell Tracer].
Antifungal activity of medicinal plants against Aspergillus niger was evaluated using Bio-Cell Tracer (BCT). By this system enabled the real time determination of hyphal growth rate could be done in the presence or absence of Chinese herbal extracts. Of the 41 herbal extracts with 60% acetone extracts tested by the present method, 26 were found to contain active components against A. niger hyphal growth. In contrast, by conventional methods, no active component was detected from every herbal extract except 3 herbal extracts. From the extract of Anemarrhena asphodeloides, one of active components was isolated and its structure determined by NMR, UV, and mass spectroscopy. The compound was identified as broussonin A (2-3-(4-hydroxyphenyl)propyl)-5-methoxy-phenol) which was formerly reported as a phytoalexin of Broussonetia papyrifera Vent.
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Utilization of excess sludge by acetone-butanol-ethanol fermentation employing Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564).
Clostridium saccharoperbutylacetonicum N1-4 could not grow or produce butanol in excess sludge medium. However, adding glucose to the excess sludge medium resulted in a specific growth rate and butanol productivity of 0.29 h(-1) and 0.55 g/l/h, respectively, and the final butanol production reached 9.3 g/l. Since the content of suspended solids in medium reduced to less than 50% of the initial content during acetone-butanol-ethanol (ABE) fermentation, the sludge was quantitatively decreased by this fermentation employing this strain.
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Substrate specificity of alpha-glucuronidase isolated from snail acetone powder.
The substrate specificity of a p-nitrophenyl alpha-D-glucopyranosyl-uronic acid-hydrolyzing enzyme (PNP-GAase) isolated from snail acetone powder has been investigated with various substrates, such as p-nitrophenyl alpha-D-glucopyranosyluronic acid (PNP-GA), 2-O-alpha- D-glucopyranosyluronic acid-D-xylose (GA-2X), 2-O-(4-O-methyl-alpha-D-glucopyranosyluronic acid)-D-xylose (MeGA-2X), and O-alpha-D-glucopyranosyluronic acid-alpha-D-glucopyranosiduronic acid (GA-GA). The Km (mM) and Vmax (mumol of glucuronic acid formed/mg of enzyme protein/min) toward these substrates were as follows; 0.13 and 3.21 for PNP-GA, 0.33 and 0.089 for GA-2X, 17.6 and 0.094 for MeGA-2X, and 0.36 and 0.015 for GA-GA, respectively. The results indicate that the PNP-GAase specifically hydrolyzed PNP-GA, however, the enzyme had broad substrate specificity.
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Total protein extraction with TCA-acetone.
We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneous precipitation and denaturation with TCA and 2ME in cold acetone. We also describe protein solubilization prior to IEF, either in classical rod gels or in IPGs, using two different solutions. The procedure is easy to carry out. The major caveats are (1) keep samples at low temperature during extraction, and then (2) manage protein samples at about 22 to 25 degrees C to avoid urea precipitation.
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Production of acetone butanol ethanol from degermed corn using Clostridium beijerinckii BA101.
In this article we report on acetone butanol ethanol (ABE) fermentation characteristics of degermed corn when using Clostridium beijerinckii BA101. Recent economic studies suggested that recovery of germ from corn and hence corn oil would help to make the ABE fermentation process more economical. C. beijerinckii BA101 ferments corn mash efficiently to produce ABE under appropriate nutritional and environmental conditions. Corn mash contains germ/corn oil that is, possibly, ancillary to the production of butanol during the ABE fermentation process. Since the presence of corn oil is not a critical factor in solvent fermentation, it can be removed and this will allow for byproduct credit. Batch fermentation of degermed corn resulted in 8.93 g/L of total ABE production as compared with 24.80 g/L of total ABE when supplemented with P2 medium nutrients. During the course of the germ separation process, corn steeping is required prior to grinding and removing the germ. It is likely that some nutrients from the corn are leached out during the steeping process. This may reduce the rate of fermentation and impact the final concentration of butanol/ABE that can be achieved. Fermentation of degermed corn with corn steep liquor resulted in the production of 19.28 g/L of ABE.
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Photodynamics of the bacteriochlorophyll-carotenoid system. 1. Bacteriochlorophyll-photosensitized oxygenation of beta-carotene in acetone.
Carotenoids are well-known physical quenchers of chlorophyll excited states and reactive oxygen species both in vivo and in vitro. They may also be involved in chemical quenching undergoing, e.g. isomerizations or oxidations. We have found that beta-carotene (Car) in aerobic acetone is rapidly oxygenated under strong illumination with red light (lambda exc > or = 630 nm) in the presence of bacteriopheophytin a. At the same time the photosensitizer undergoes only slight (< 10%) photodegradation. By preparative high-performance liquid chromatography as many as seven major products of oxygen attachment to Car have been isolated. Their molecular masses show that Car sequentially accumulates up to six oxygen atoms while its C40-skeleton remains intact.
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Acetone-dependent regulation of cytochrome P-450j (IIE1) and P-450b (IIB1) in rat liver.
1. Concomitant changes in the concentration of P450 IIB1 (P-450b) and the rate of O-depentylation of 7-pentoxyresorufin was observed in rat liver microsomes after a single intragastric dose of acetone. 2. In contrast the concentration of P450 IIE1 (P-450j) did not coincide with changes in the rate of P450 IIE1-dependent p-nitrophenol hydroxylation or metabolic activation of carbon tetrachloride. 3. Quantification of the proteins in lysosomes indicated that both P450 IIB1 and P450 IIE1 are degraded via an autophagosomal/autolysosomal pathway. 4. It is concluded that P450 IIE1 is catalytically inactivated in microsomes prior to the degradation of this protein.
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