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Metabolism of N-nitroso-1,3-thiazolidine in the rat.

[2-14C]N-nitroso-1,3-thiazolidine (125 mg/kg body weight) administered by gavage to rats was excreted mostly (81% of the dose) in the urine with a smaller amount (7%) in respired air. The major metabolite in the urine (77.2%) was 2-hydroxy-1,3-thiazolidine and minor quantities of the isomers 4- and 5-hydroxy-1,3-thiazolidine were also detected. The presence of hydroxy-1,3-thiazolidine, determined using mass spectrometry and high-performance liquid chromatography, indicated that alpha-hydroxylation and denitrosation are major metabolic pathways of N-nitroso-1,3-thiazolidine. Analysis of the non-acidic urine extract with gas chromatography-mass spectrometry (GC-MS) using untreated on-column injection and heated split injection gave differing results. Compounds detected using the latter technique may have been artefacts formed by pyrolysis in the heated injection port. Analysis of the metabolites by direct probe mass spectrometry (unheated) and Fourier transform infra-red spectroscopy supported the results seen with on-column GC-MS.

Animals↗

Modified mutagen activation in hepatic fractions from rats fed dietary rutin--interaction between gut flora and host metabolism.

Male Sprague-Dawley rats were fed a purified diet or one supplemented with the glycosidic plant flavonoid (+)rutin for 14 days. Rutin treatment significantly increased caecal bacterial beta-glucosidase activity (responsible for the conversion of rutin to the flavonoid quercetin) and there was an associated increase in the capacity of hepatic fractions (S-9) to activate the food pyrolysis products IQ, MeIQ and MeIQx to bacterial mutagens in vitro. Hepatic conversion of aflatoxin B1 to a mutagen was unaltered while in vitro activation of quercetin was significantly lower in tissue fractions from the rutin-fed rats compared with those from controls. Rutin treatment was without effect, however, on a number of hepatic cytochrome P-450-dependent mixed-function oxidase activities. The results suggest that products of bacterial metabolism of rutin formed in the hindgut may influence the activity of hepatic enzymes involved in the activation of certain classes of mutagen.

Animals↗

Applications of the in vitro aryl hydrocarbon hydroxylase induction assay for determining "2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents": pyrolyzed brominated flame retardants.

The pyrolysis of brominated flame retardants FR 300 BA (decabromobiphenyl) ether, FireMaster BP-6 (polybrominated biphenyls), Bromkal 70-5-DE (primarily pentabromodiphenylether), Bromkal 70-DE (primarily penta and tetrabromodiphenylether) and Bromkal G1 (pentabromodiphenylether) resulted in the formation of relatively high levels of polybrominated dibenzofurans (PBDFs) and dibenzo-p-dioxins (PBDDs as determined by gas chromatography-mass spectrometric analysis. The dose response EC50 values for the induction of aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase (EROD) by the flame retardant pyrolysates was determined in rat hepatoma H-4-II E cells and compared to the relative induction activities of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the concentrations of "2,3,7,8-TCDD equivalents" were calculated. The range of "2,3,7,8-TCDD equivalents" levels (micrograms/g or ppm) derived from values obtained from the AHH and EROD bioassays for each of the pyrolyzed flame retardant samples was: 174-194, 480-1400, 2140-4680, 6740-8780 and 3920-5260 ppm for FR 300 BA, FireMaster BP-6, Bromkal 70 DE, Bromkal 70-5 DE and Bromkal G1, respectively. The in vivo dose-response effects of 2 pyrolyzed flame retardants were determined in immature male Wistar rats and compared to the dose-response activities of 2,3,7,8-TCDD. The in vivo responses which were measured included hepatic microsomal AHH and EROD induction, body weight loss and thymic atrophy. For the pyrolyzed FireMaster BP-6 and Bromkal 70-5 DE samples, the range of calculated in vivo "2,3,7,8-TCDD equivalents" (ppm in sample) for the 4 in vivo bioassays was 520-1780 ppm and 3860-8960 ppm, respectively. The excellent overlap between the in vivo and in vitro 2,3,7,8-TCDD equivalents for the 2 flame retardant pyrolysate extracts supports the utility of the in vitro induction bioassay for quantitatively determining "2,3,7,8-TCDD equivalents" for mixtures containing toxic halogenated aryl hydrocarbons.

Animals↗

Formation of mutagens in cooked foods. III. Isolation of a potent mutagen from beef.

The major mutagenic component of fried beef has been isolated using a series of chromatographic steps. The pure compound has been analyzed by low and high resolution mass spectroscopy and nuclear magnetic resonance spectroscopy. The results indicate that the molecular weight of this extremely mutagenic compound is 198, with an elemental composition of C11H10N4. The compound is different from the known mutagenic pyrolysis products of amino acids or proteins.

Animals↗

Induction of 8-azaguanine or ouabain resistant somatic mutation of Chinese hamster lung cells by treatment with tryptophan products.

The basic fraction of tryptophan pyrolysis products (TBF) showed strong mutagenic activity on somatic cells of the lung of Chinese hamsters. In this somatic mutation test, TBF was demonstrated to have 5.6 times higher mutagenicity than diethylnitrosamine (DEN) when mutants were selected with 8-azaguanine, and 13.5-fold higher mutagenicity than DEN when mutants were selected with ouabain. From these findings, it is suggested that pyrolyzates of amino acids may have mutagenic actions on somatic cells of animals, as well as carcinogenic actions.

Animals↗

Detection of potent mutagens, Trp-P-1 and Trp-P-2, in broiled fish.

The potent mutagens Trp-P-1 (3-amino-1, 4-dimethyl-5H-pyrido-[4,3-b]-indole) and Trp-P-2 (3-amino-1-methyl-5H-pyrido[4,3-b]indole) are known to be produced by pyrolysis of tryptophan [8]. To determine whether such mutagens are produced by cooking foods, the fractions obtained from broiled sardines cooked in the ordinary way were analysed by gas chromatography/mass spectrometry. The results showed that 13.3 ng of Trp-P-1 and 13.1 ng of Trp-P-2 were, in fact, present per gram of broiled sardines.

Animals↗

Metabolic formation of pyrenequinones as enhancing agents of mutagenicity in Salmonella.

Pyrene and 1-methylpyrene in the pyrolysis product of cellulose, which enhances mutagenicity of 2-acetylaminofluorene, must be metabolized to expose enhancing activity. 1,6-Pyrenequinone and 1,8-pyrenequinone, having high enhancement activity, were first isolated and identified in the metabolites of pyrene by rat liver microsomal fraction.

2-Acetylaminofluorene↗

Mutagenicity of pyrolysates of salt-tolerant bacteria from food and cigarettes.

Smoke condensates obtained by pyrolysis of the cells of microorganisms isolated from food and cigarettes and of laboratory cultures were shown to be mutagenic to Salmonella typhimurium TA98 and TA100 in incorporation with liver microsomal fraction S-9 mix. The smoke condensates from salt-tolerant bacteria, which belonged to Micrococcus, Bacillus and Corynebacterium spp., showed higher mutagenic activity than those from other less salt-tolerant organisms, fish flesh or pork. Smoke deriving from microorganisms, especially salt-tolerant ones, in food or cigarettes as well as from food components might take part in human carcinogenesis because of the correlation between mutagenicity and carcinogenicity.

Bacteria↗

Recent methods for the determination of volatile and non-volatile organic compounds in natural and purified drinking water.

Four analytical protocols for the extraction and preconcentration of organic residues in natural or purified drinking water were investigated and compared: closed loop stripping analysis; simultaneous extraction-distillation; purge and trap analysis; continuous liquid-liquid extraction. Organic extracts were submitted to a variety of separation and identification techniques. Volatiles were determined by conventional capillary column gas chromatography with tandem mass spectrometry, using triple-stage quadrupole instruments. Non-volatile and thermally labile molecules were investigated by several different techniques (high-temperature gas chromatography, capillary column supercritical fluid chromatography, pyrolysis gas chromatography-mass spectrometry, thermospray liquid chromatography with tandem mass spectrometry and conventional fast-atom bombardment with tandem mass spectrometry). Several samples recently examined in the laboratory provide examples of this multitechnique approach for a more complete knowledge of the organic carbon distribution in water-dissolved organic matter, taking into account organic substances with widely different volatilities, polarities and thermal stabilities.

Chlorides↗

Gas chromatographic-mass spectrometric method for determination of anabasine, anatabine and other tobacco alkaloids in urine of smokers and smokeless tobacco users.

A selected ion monitoring method for determination of the tobacco alkaloids anabasine, anatabine, nornicotine, metanicotine, dihydrometanicotine, and 2,3'-bipyridyl in urine of smokers and smokeless tobacco users is described. The method involves conversion of the secondary amine alkaloids to tertiary amine derivatives by reductive alkylation using an aldehyde and sodium borohydride, and chromatography on a 5% phenylmethylsilicone capillary column. These derivatives have good chromatographic properties, allowing determination of concentrations as low as 1 ng/ml. The alkaloid 2,3'-bipyridyl is unaffected by the derivatization procedure and may be determined simultaneously with the other alkaloids. The structural analogues 2-(3-pyridyl)hexahydroazepine, 5-methyldihydrometanicotine, and 6-methyl-2,3'-bipyridyl were synthesized for use as internal standards. Using the method, concentrations and 24 h excretion of anabasine, anatabine, and nornicotine in urine of twenty-two smokers, eight chewing tobacco users, and six oral snuff users were determined and compared with concentrations and excretion of nicotine and its metabolite cotinine. Excretion of nicotine and cotinine was similar in all tobacco users, but excretion of anabasine, anatabine and nornicotine was substantially greater in urine of smokeless tobacco users, presumably due to absence of pyrolysis of these alkaloids in smokeless tobacco products.

Alkaloids↗

Determination of manganese in human brain samples.

A method is presented for the determination of manganese (Mn) in human tissue samples (especially brain) by graphite furnace atomic absorption spectrophometry (GFAAS). After complete digestion by a mixture of concentrated nitric acid (HNO3)/concentrated perchloric acid (HClO4) (50:50, v/v), the samples are assayed on a Perkin-Elmer 5100 PC apparatus, equipped with transversal graphite tubes and a Mn-specific hollow cathode lamp. The furnace conditions are as follows (for each step: temperature (degree C)/ramp (s)/duration (s)) dry 120/1/40; char 1200/5/10; atomization 2250/0/4; pyrolysis 2400/1/1. Zeeman correction is employed. The method is linear over the range 0.05 to 5.00 micrograms/g wet tissue, and the limit of detection for Mn is about 0.01 microgram/g wet tissue. This simple and rapid method may be of value for the post-mortem assessment of Mn accumulation in brain structures due to occupational or iatrogenic exposure. An application is presented in which elevated levels of Mn were determined in the brain samples of a 63-year-old female deceased after long-term total parenteral nutrition involving Mn supplementation.

Brain Chemistry↗

Simple methods for the qualitative identification and quantitative determination of macrolide antibiotics.

Pyrolysis-gas chromatography is shown to be a rapid straightforward method for the qualitative differentiation of the macrolide antibiotics erythromycin, oleandomycin, troleandomycin, spiramycin and tylosin. Organic salts do not interfere and identification of erythromycin and troleandomycin in commercial products is viable. Spectrophotometric quantitation of these same five antibiotics after reaction with concentrated sulphuric acid is studied at about 470 nm. Reaction conditions such as acid concentration, time and temperature are provided. The sugar moieties of the antibiotics are proposed as the reactive sites. Detection limits are about 0.2-1.0 microg ml-1 [corrected] and analysis of pharmaceutical products should be possible.

Anti-Bacterial Agents↗

Peroxyl radical formation in aqueous solutions of N,N-dimethylformamide, N-methylformamide, and dimethylsulfoxide by ultrasound: implications for sonosensitized cell killing.

Sonodynamic therapy, which refers to a synergistic effect of drugs and ultrasound, is a promising new modality for cancer treatment. The sonodynamic effect was found for a number of structurally unrelated compounds, and the underlying mechanisms are still unknown. Recently, Jeffers et al. (J. Acoust. Soc. Am. 97:669-676; 1995) have shown that the sonodynamic action of nontoxic concentrations of N,N-dimethylformamide (DMF), N-methyl formamide (MMF), and dimethylsulfoxide (DMSO) combined with ultrasound, on killing of cultured HL-60 human promyelocytic leukemia cells, and attributed this toxic effect to unknown short lived reactive species produced from these solutes by ultrasonic cavitation. Using the spin trap 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) in nitrogen-saturated aqueous solutions of DMF, MMF, or DMSO exposed to 50 kHz ultrasound, we detected formation of .CH3 and .CH2N(CH3)CHO radical adducts for DMF, mostly .CH2NHCHO adducts for MMF, and .CH3 adducts for DMSO. These radicals were formed either by reactions of the solutes with ultrasound-generated .H and .OH radicals (such as .CH2R-type radicals in DMF and MMF, and .CH3 radicals in DMSO), or by direct pyrolysis of the weak bonds in the solute molecules (e.g., .CH3 radicals from DMF). In air-saturated sonicated solutions these carbon centered radicals were converted to the corresponding peroxyl radicals and spin trapped with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO); .OOCH2N(CH3)CHO radicals were identified in DMF, .OOCH2NHCHO radicals in MMF, and .OOCH3 radicals in DMSO solutions. We suggest that these radical species by virtue of their longer lifetimes and higher selectivity, compared to .OH radicals, which are also formed in sonicated solutions, are the species responsible for sonodynamic cell killing by the combined effect of ultrasound with DMF, MMF, or DMSO.

Benzenesulfonates↗

The chemical characterization of melanin contained in substantia nigra of human brain.

The pigment of substantia nigra human brain has been extracted by a mild procedure consisting of washes with phosphate buffer, methanol and incubation with SDS-proteinase. Pyrolysis gas chromatography mass spectrometry, infrared spectrometry, termogravimetric analysis and elemental analysis were the techniques used for the chemical characterization. An indole moiety bound to a sulfur containing amino acid and to palmitic acid were the main aspects found in the structure. The presence of a 7% inorganic component was observed. This probably contains Fe, Cu, Zn and Cr which are also relevant, for the formation and the role of melanin in substantia nigra neurons. The fatty acid moiety is chemically bound to the indole structure as it was not eliminated by repeated methanol washing. The same situation occurs for the sulfur containing group. Considering these data and the most abundant molecules present in substantia nigra the precursor of neuromelanin seems to be a cysteinyl-catechol, to which is then bound a palmityl group.

Adult↗

The resistance to carbon dioxide laser ignition of a new endotracheal tube: Xomed Laser-Shield II.

STUDY OBJECTIVE: To assess the resistance of a new endotracheal tube to penetration and ignition by a carbon dioxide (CO2) laser at a clinically relevant power setting. DESIGN: An unblinded, in vitro trial simulating clinical conditions. SETTING: The operating room at a university medical center. PATIENTS: None. INTERVENTIONS: A CO2 laser at 20W with a beam diameter of 1 mm was directed onto the Xomed Laser-Shield II at several points: both perpendicular and tangential to the portion of the tube covered by the protective wrap, to the unwrapped silicone tubing just proximal to the cuff, to the cuff filled with saline and methylene blue, and to the unwrapped silicone tubing distal to the cuff. Trials were performed with or without supplemental oxygen (O2) blown onto the study site and with a 6 L/min flow through the tube containing one of four concentrations of O2 in O2-air admixtures: 21%, 30%, 40%, and 100%. MEASUREMENTS AND MAIN RESULTS: Under all conditions, the laser did not penetrate the protective wrap, but the overlying polytetrafluoroethylene (Teflon) sheet was vaporized to an area equal to the size of beam application. The cuff was penetrated by the laser within 5 seconds. The unprotected silicone protein of the tube was penetrated by the laser in approximately 5 seconds, and time to formation of a blowtorch phenomenon shortened with increasing the percentage of O2 flowing through the tube. CONCLUSIONS: The protective coating of the Xomed Laser-Shield II is laser-resistant, but the unprotected silicone proximal and distal to the cuff is laser-vulnerable and can, under certain conditions, promote a blowtorch phenomenon. This tube would be acceptable for use in oral and pharyngeal laser surgery, but we recommend its use only in well-ventilated areas, out of concern for exposure to the products of the pyrolysis of Teflon, specifically the development of polymer-fume fever.

Equipment Design↗

The effects of ultraviolet light on the degradation of organic compounds: a possible explanation for the absence of organic matter on Mars.

The analysis of the top layer of the Martian regolith at the two Viking landing sites did not reveal any indigenous organic compounds. However the existence of such compounds at deeper layers cannot be ruled out. Cosmochemical considerations indicate various potential sources for organic matter on Mars, such as comets and meteorites. Its disappearance from the top layer could be caused by degradation processes on the surface of the planet. Possible destructive agents include ultraviolet light, oxygen and metal oxides. In this study we tested the stability of a sample of the Murchison meteorite and various organic substances which have been detected in carbonaceous chondrites, such as glycine, adenine and naphthalene, to the action of ultraviolet light. The compounds were adsorbed on powdered quartz and on California desert soil and were irradiated in the presence or absence of oxygen. The organic content, before and after irradiation, was measured by carbon elementary analysis, UV-absorption, amino acid analysis or pyrolysis-gas chromatography-mass spectrometry. In the absence of oxygen, adenine and glycine appear to be stable over the given period of irradiation. A definite degradation was noticed in the case of naphthalene and the Murchison meteorite. In the presence of oxygen in amounts comparable to those on Mars all compounds were degraded. The degree of degradation was influenced by the irradiation time, temperature and oxygen content.

Adenine↗

Silicon oxycarbide glasses for blood-contact applications.

Silicon oxycarbide (SiOxCy) glass compositions are shown to exhibit a variable propensity to contact activate coagulation of whole human blood plasma that depends on X:Y surface stoichiometry. SiOxCy exhibit activation properties similar to pyrolytic carbon (PC) over a broad range of X:Y ratios. Surface composition of SiOxCy glass powders prepared by pyrolysis of thermosetting polysilsequioxanes roughly correlated with total carbon concentration of precursor resins and could be significantly modified by etching in alkaline solutions. Results suggest that SiOxCy may offer unique properties as a substitute for PC in medical-device applications demanding excellent tribological properties, such as artificial heart valves.

Biocompatible Materials↗

Novel technology for rapid species-specific detection of Bacillus spores.

There is an urgent need for a small, inexpensive sensor that can rapidly detect bio-warfare agents with high specificity. Bacillus anthracis, the causative agent of anthrax, would be a perilous disease-causing organism in the event of a release. Currently, most anthrax detection research is based on nucleic acid detection, immunoassays and mass spectrometry, with few detection levels reported below 10(5) spores. Here, we show the ability to distinguish Bacillus spores to a level approaching 10(3) spores, below the reported median infectious dose of B. anthracis, using pyrolysis--micromachined differential mobility spectrometry and novel pattern recognition algorithms that combine lead cluster mapping with genetic algorithms.

Algorithms↗