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Asthma in children exposed to nitrogen dioxide in ice arenas.

Very high concentrations of nitrogen dioxide (NO2) have been measured in arenas using combustion engine-powered resurfacing machines. This study was performed to compare the occurrence of asthma in children playing ice hockey in arenas using propane-powered machines and in children attending arenas using electric machines. Children regularly playing hockey in the arenas (nine propane, six electric) were sent a questionnaire, including questions on allergic disease and risk factors. Measurements of NO2 were performed with passive diffusion samplers during 3 consecutive days. The mean NO2 concentration in the propane arenas was 276 microg x m(-3) (range 28-1015 microg x m(-3)) and 11 microg x m(-3) (2-30) in the electric arenas. Questionnaires were answered by 1,536 children (78%), with an overall prevalence of asthma of 16%. The odds ratio (OR) for asthma was 0.9 (95% confidence interval (CI) 0.7-1.2) comparing propane arenas to electric. However, children in propane arenas with higher than median concentration of NO2 reported more wheezing (OR 1.4, 95% CI 1.0-1.9) and nasal symptoms (OR 1.7, 95% CI 1.3-2.3) than children in propane arenas with lower concentrations. In conclusion, children playing ice hockey in indoor arenas have a high prevalence of asthma, but it appears unlikely that increased exposure to combustion products, including nitrogen dioxide, is a major contributor to this excess risk.

Adolescent↗

The metabolism of 3-methoxyamphetamine in dog, monkey and man.

1. The metabolism of 3-methoxyamphetamine in vivo was examined in dog, monkey and man. 2. The metabolites identified in all three species were 3-O-methyl-alpha-methyldopamine, 1-(3-methoxy-4-hydroxyphenyl)propan-2-ol, 3-hydroxyamphetamine, 1-(3-hydroxyphenyl)propan-2-one, 1-(3-hydroxyphenyl)propan-2-ol, 1-(3-methoxyphenyl)propan-2-ol and 1-(3-methoxyphenyl)propane-1,2-diol. 3. 1-Hydroxyl-1(3-hydroxyphenyl)propan-2-one was tentatively identified in the urine of all three species. 4. 4-O-Methyl-alpha-methyldopamine was also found in the urine of dog and monkey but not in human urine.

Ammonium Hydroxide↗

[Study on the phase I metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD].

AIM: To study the phase I metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. METHODS: DDPH was administered i.p. to bile duct-cannulated rats. Bile samples were collected before administration and up to 12 h after administration. After being treated with beta-glucuronidase, the bile samples were purified and enriched with C-18 SPE columns, and then were analyzed by LC/DAD/MSD. The samples containing synthesized reference standards of DDPH metabolite 1-(2, 6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethylamino)-propane (M1), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxy-phenylethylamino)-propane (M2), 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3,4- methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3-hydroxy-4- methoxyphenylethylamino)-propane (M4), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2- (3-hydroxy-4-methoxyphenylethylamino)-propane (M5) and 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3-methoxy-4- hydroxyphenylethylamino)-propane (M6) were analyzed by LC/DAD/MSD under identical conditions. RESULTS: The retention times, UV spectra, molecular weights and production spectra (obtained by collision-induced dissociation) of the apparent ions of peak A, B, C, D, E and F in the total ion chromatogram of DDPH treated rat bile sample were consistent with those of M1, M2, M3, M5, M4 and M6, respectively. CONCLUSION: M1, M2, M3, M4, M5 and M6 were identified as the phase I metabolites of DDPH in the rat.

Animals↗

Temperature dependence of mature mouse oocyte membrane permeabilities in the presence of cryoprotectant.

Knowledge of cell membrane permeability characteristics facilitates the design of cryopreservation protocols which minimize damage from osmotic stress and reduce the incidence of intracellular freezing. Such permeability characteristics can be determined for oocytes from volume measurements taken during exposure to cryoprotectant. Individual mouse oocytes were held using negative pressure applied to the zona pellucida by means of a micropipet. Each oocyte was perfused with 1 ml 1.5 mol liter-1 dimethyl sulfoxide (Me2SO) or propane-1,2-diol at 30, 23, or 10 degrees C. The osmotic response of each oocyte before, during, and after perfusion was recorded by videomicroscopy until equilibrium was reached. Mean cell diameter across three axes was used to calculate oocyte volume, assuming sphericity, and, using mathematical modeling, values for hydraulic conductivity (Lp) were found to be 0.64, 0.41, and 0.20 micron min-1 atm-1 in the presence of Me2SO and 0.53, 0.36 and 0.15 in the presence of propane-1,2-diol at 30, 23, and 10 degrees C, respectively. Cryoprotectant permeability (omega) was 0.37, 0.16, and 0.035 for Me2SO and 0.43, 0.24, and 0.04 for propane-1,2-diol, while the reflection coefficient was 0.98, 0.94, and 0.99 (Me2SO) and 0.76, 0.99, and 0.95 (propane-1,2-diol) all at 30, 23, and 10, respectively. The corresponding activation energies (Ea) were 11.65 and 12.23 kCal mol-1 for Lp and 23.52 and 22.48 kCal mol-1 for omega, in the presence of Me2SO and propane-1,2-diol, respectively. Values generated for Lp and associated Ea were similar to those found for mouse oocytes in the absence of cryoprotectant, while omega and its Ea were similar to those found for oocytes of other species.

Animals↗

Pentane-1,5-diol as a percutaneous absorption enhancer.

Propylene glycol (propane-1,2-diol) is the only diol widely used in dermatology. Pentane-1,5-diol is mainly used as a plasticizer in cellulose products and adhesives, in dental composites and in brake fluid compositions and as a preservative for grain. However, pentane-1,5-diol is also an effective solvent, water-binding substance, antimicrobial agent and preservative and may therefore replace several ingredients in a skin composition. The release of tri-iodothyroacetic acid (TRIAC) and percutaneous absorption of hydrocortisone and mometasone furoate with either pentane-1,5-diol or propane-1,2-diol and 2-methyl-pentane-2,4-diol (hexylene glycol), respectively, as enhancers was compared. The release of TRIAC was 21% higher when pentane-1,5-diol was used as an enhancer instead of propane-1,2-diol. The percutaneous absorption of hydrocortisone through the skin was increased 12 times with propane-1,2-diol compared to 4.4 times with pentane-1,5-diol. However, the percutaneous absorption of hydrocortisone into the skin was 50% higher with pentane-1,5-diol compared to propane-1,2-diol. There was no significant difference, between the original mometasone furoate cream, with 2-methyl-pentane-2,4-diol, and the new cream with pentane-1,5-diol in the amount of mometasone furoate that was absorbed into the skin and through the skin. However, the cosmetic properties of the new mometasone furoate cream was superior to the original mometasone furoate cream, for examples, no bad odour, more even texture, goes better into the skin and has less greasiness. Pentane-1,5-diol can be used as a technology platform, which adds a series of desirable properties to dermatological preparations and enhances product usability. This will result in improved formulations for a series of major and commonly used dermatological drugs. When used in pharmaceutical topical preparations, pentane-1,5-diol will increase the percutaneous absorption of the active substance and it is an efficient antimicrobial agent that will act as an effective preservative in topical formulations. Pentane-1,5-diol is cosmetically attractive, has low risk for skin and eye irritation compared to other diols, low toxicity risk and no bad odour.

Administration, Cutaneous↗

Effect of a 1 min hand wash on the bactericidal efficacy of consecutive surgical hand disinfection with standard alcohols and on skin hydration.

BACKGROUND: In most surgical theatres, a 1 min or even longer hand wash is routine as part of the pre-operative hand disinfection. But its benefit has recently been seen critically. METHODS: We have therefore investigated the effect of a 1 min hand wash on skin hydration and on the efficacy of consecutive surgical hand rubbing with three standard alcohols (60% propan-1-ol, 60% propan-2-ol, 80% ethanol; all v/v) on the resident hand flora. Three types of treatment were performed: (i) a 1 min pre-wash before surgical hand disinfection, (ii) no pre-wash before surgical hand disinfection and (iii) no pre-wash but use of a brush for 1 min during disinfection procedure. The efficacy of the alcohols was determined according to prEN 12791 with the same 20 volunteers in paired groups. To assess the effect of the hand wash on skin hydration, 10 volunteers washed their hands with sapo kalinus for 1 min and dried hands with a paper towel. Skin hydration was measured with a corneometer before the hand wash and subsequently up to 10 min thereafter both on the palm and dorsum of hands. We also tested the reduction of bacterial spores by a 15 s hand wash according to EN 1499 after artificial contamination of hands of 14 volunteers with spores of B. stearothermophilus. RESULTS: Propan-1-ol (60%) was most effective with a mean log10 reduction of 2.11, followed by ethanol (80%) with a mean log10 reduction of 1.76 and propan-2-ol (60%) with a mean log10 reduction of 0.57 (all immediate effect without hand wash). The efficacy of the alcohols was neither significantly improved nor impaired by a preceding 1 min hand wash, but there is a trend towards better efficacy on dry hands. Using a brush for 1 min during disinfection resulted in a better efficacy with all alcohols. An anaylsis of variance revealed that the immediate effect of ethanol (p = 0.013) and propan-2-ol (p = 0.001) is significantly influenced by the variation of treatments which is mainly explained by the effect of brushing during disinfection. But no significant difference between treatment variations was found in the sustained effect with any of the alcohols. Skin hydration increased significantly by a 1 min hand wash for up to 10 min despite drying hands with a paper towel. A 15 s hand wash reduced the number of bacterial spores significantly from log10 3.84 to log10 1.99 (p = 0.001). CONCLUSIONS: There is no benefit of a hand wash as part of surgical hand disinfection except that a short hand wash of 15 s can effectively reduce spores. The best time for this short hand wash is at the beginning of work in hospital, but at the latest in the sluice of the operating theatre about 10 min before applying an alcohol-based hand rub to give the skin enough time to dry.

Adult↗

Efficacy of three ethanol-based hand rubs against feline calicivirus, a surrogate virus for norovirus.

We investigated the efficacy of three ethanol-based hand rubs (Sterillium Virugard, 95% ethanol; Sterillium Rub, 80% ethanol; Desderman N, 75.1% ethanol) against feline calicivirus (FCV), the surrogate virus for norovirus, on artificially contaminated hands of healthy volunteers. The ASTM E 1838-02 standard was used. Experiments were controlled with 70% ethanol and 70% propan-1-ol which were previously found to have maximal efficacy against FCV. In the first step, three different organic loads (5% fetal bovine serum, 5% faecal suspension and the tripartite ASTM load) were compared. A significant influence of the type of organic load was found (P<0.001, ANOVA). In the second step, the hand rubs were investigated with a 5% faecal suspension as a challenging organic load. The hand rub based on 95% ethanol was more effective than those based on 70% ethanol (mean log10 reduction factor: 2.17 vs. 1.56; P=0.17) and 70% propan-1-ol (mean RF: 1.63 vs. 0.95; P=0.0003). The hand rub based on 80% ethanol was also more effective than those based on 70% ethanol (mean RF: 1.25 vs. 1.03: P=0.20) and 70% propan-1-ol (mean RF: 1.43 vs. 1.09; P=0.03). The hand rub based on 75.1% ethanol was less effective than those based on 70% ethanol (mean RF: 1.07 vs. 1.27; P=0.47) and 70% propan-1-ol (mean RF: 0.78 vs. 0.97; P=0.35). Based on our data, ethanol has superior efficacy against FCV than propan-1-ol. In addition, a higher ethanol concentration in three commercially available hand rubs was associated with better efficacy against FCV.

1-Propanol↗

Adaptation of the membrane fatty acid composition by growth in the presence of n-alkanols influences glycosyltransferase expression in Streptococcus salivarius.

Growth of Streptococcus salivarius ATCC 25975 in the presence of n-alkanols in the series methanol to decan-1-ol led to a decrease in the unsaturated to saturated fatty acid ratio. Each member of the set of n-alkanols which was examined over a range of concentrations possessed a point at which extracellular glucosyltransferase (GTF) production was minimal; increasing the concentration of the n-alkanol past this point stimulated GTF production. This effect was greatest with hexan-1-ol although it was observed to a lesser extent with pentan-1-ol and heptan-1-ol. Reduced cell-associated fructosyltransferase activity was observed with increasing concentrations of each n-alkanol. Growth in the presence of 25 mM-propan-1-ol gave rise to a fatty acid profile in which 55% of the fatty acids were of an odd chain length. S. salivarius ATCC 25975 was shown to be able to utilize ethanol in a similar manner to propan-1-ol by growing it in the presence of 400 mM-[14C]ethanol. Analysis of the membrane lipids at the stationary phase of growth indicated that 17.6% of the carbon of the fatty acids was derived from ethanol. A leaky adh mutant, S. salivarius MJ 37501, was isolated. The leaky nature of the mutant enabled it to incorporate reduced levels of odd-chain-length fatty acids into its membrane lipids when grown in the presence of 100 mM-propan-1-ol, but not when grown in the presence of 25 mM-propan-1-ol. S. salivarius ATCC 25975 therefore metabolized propan-1-ol (and ethanol) via a constitutive alcohol dehydrogenase.

Adaptation, Biological↗

Effects of ethanol and other aliphatic alcohols on NO-mediated relaxations in rat anococcygeus muscles and gastric fundus strips.

1. In anococcygeus muscles, ethanol (20-500 mM) slightly increased the tone and inhibited relaxations elicited by nitrergic nerve stimulation (0.5-5 Hz) in a concentration-dependent manner. 2. Other aliphatic alcohols decreased the tone but had inhibitory effects similar to ethanol on stimulation-induced relaxations, the EC50 (mM) values being: methanol 280, ethanol 80, propan-1-ol 20, propan-2-ol 55, propan 1,2-diol 135, butan-1-ol 120, butan-2-ol 15 and pentan-1-ol 3. 3. Relaxations induced by sodium nitroprusside (SNP, 10 nM) were inhibited by ethanol (20-500 mM) in a concentration-dependent manner and by propan-2-ol (100 mM). Relaxations induced by NO (1 microM) were inhibited by high concentrations of ethanol (200-300 mM) and by propan-2-ol (100 mM). 4. In gastric fundus strips, ethanol (60-200 mM) did not affect the resting tone but inhibited NO-mediated relaxations elicited by low frequency (1 Hz) field stimulation and reduced the initial relaxation by high frequency field stimulation (10 Hz) and by SNP (50 nM). The relaxant action of isoprenaline (10 nM) was not reduced although it was slightly slower in onset. Other aliphatic alcohols tested decreased the tone and inhibited relaxations elicited by field stimulation. 5. Acetaldehyde (1-10 mM) inhibited relaxations elicited by field stimulation and SNP in both the rat anococcygeus muscles and gastric fundus strips. The tone of gastric fundus strips was decreased by acetaldehyde but it was transiently increased in anococcygeus muscles. 6. The dehydrogenase inhibitors, 4-methylpyrazole (100 MicroM) and disulfiram (100 MicroM) did not significantly affect the inhibition by ethanol of nitrergic nerve stimulation-induced relaxations in anococcygeus muscles, which suggests that ethanol acts directly.7. The inhibition of NO-mediated relaxations by alcohols is attributed to sequestration of NO to form nitroso-alcohols.

Acetaldehyde↗

The sensitizing potential of di-(meth)acrylates based on bisphenol A or epoxy resin in the guinea pig.

Most composite materials in dentistry used today, contain resins based on dimethacrylates. BIS-GMA [2,2-bis-(4-(2-hydroxy-3-methacryloxypropoxy)phenyl)propane], the addition reaction product of bisphenol A and glycidyl methacrylate or an epoxy resin and methacrylic acid, is used most extensively. More recently, dimethacrylates based on bisphenol A, with various chain lengths have appeared on the market as a substitute for or in addition to BIS-GMA. Such compounds are BIS-MA [2,2-bis-(4-(methacryloxy)phenyl)propane], BIS-EMA [2,2-bis-(4-(2-methacryloxyethoxy)phenyl)propane] and BIS-PMA [2,2-bis-(4-(3-methacryloxypropoxy)phenyl)propane]. Increasing interest in the radiaton cure of coatings and printing inks have focused attention on these substances and on epoxy diacrylates as radiation-curable resins. The sensitizing capacity of the different acrylates based on bisphenol A or epoxy resin have been investigated with the guinea pig maximization test. The pattern of simultaneous reactivity of the compounds was also studied. Epoxy diacrylate [2,2-bis-(4-(2-hydroxy-3-acryloxy-propoxy)phenyl)propane], BIS-EMA and BIS-MA are shown to be strong sensitizers, while the linear fraction of BIS-GMA and its isomers and BIS-PMA have none or a low sensitizing capacity. The impurities in the BIS-GMA and BIS-MA batches seem to have high allergenic potential. These results stress the importance of a pure substance when discussing allergenicity and cross reactions.

Acrylates↗

Push-pull tests for assessing in situ aerobic cometabolism.

Three types of single-well push-pull tests were developed for use in assessing the feasibility of in situ aerobic cometabolism of chlorinated aliphatic hydrocarbons (CAHs). These included transport tests, biostimulation tests, and activity tests. Transport tests are conducted to evaluate the mobility of solutes used in subsequent tests. These included bromide or chloride (conservative tracers), propane (growth substrate), ethylene, propylene (CAH surrogates), dissolved oxygen (electron acceptor), and nitrate (a minor nutrient). Tests were conducted at an experimental wellfield of Oregon State University. At this site, extraction phase breakthrough curves for all solutes were similar, indicating apparent conservative transport of the dissolved gases and nitrate prior to biostimulation. Biostimulation tests were conducted to stimulate propane-utilizing activity of indigenous microorganisms and consisted of sequential injections of site ground water containing dissolved propane and oxygen. Biostimulation was detected by the increase in rates of propane and oxygen utilization after each injection. Activity tests were conducted to quantify rates of substrate utilization and to confirm that CAH-transforming activity had likely been stimulated. In particular, the transformation of injected CAH surrogates ethylene and propylene to the cometabolic byproducts ethylene oxide and propylene oxide provided evidence that activity of the monooxygenase enzyme system, responsible for aerobic cometabolic transformations of CAHs, had likely also been stimulated. Estimated zero-order transformation rates decreased in the order propane > ethylene > propylene. The series of push-pull tests developed and field tested in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ aerobic cometabolism of CAHs.

Bacteria, Aerobic↗

Cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum: effect of surfactants and organic solvents on activity.

We have studied systematically the effect of the non-ionic surfactants Thesit and Triton X-100, and of propan-2-ol (used as a substrate solubilizer) on the activity of the cholesterol oxidases from Streptomyces hygroscopicus (SCO) and Brevibacterium sterolicum (BCO). Low concentrations of Thesit lead to an activity increase with both enzymes; at higher surfactant concentrations the opposite effect occurs. Triton X-100 inactivates both enzymes at all concentrations. It is deduced that these surfactants exert their effects by interaction with the enzymes and not by affecting micellar phenomena. The effect of propan-2-ol on SCO, in contrast with that on BCO, depends on the buffer concentration (potassium phosphate). Other organic solvents induce results similar to those obtained with SCO and propan-2-ol. A significant difference between the two cholesterol oxidases emerges when stability is tested at 25 degrees C and in the presence of different concentrations of propan-2-ol: BCO activity is rapidly inactivated, whereas SCO still has 70% of the initial activity after 5 h in the presence of 30% propan-2-ol. From our results, SCO seems to be the catalyst of choice in comparison with BCO for the exploitation of cholesterol oxidases in biotechnology and applied biochemistry.

Brevibacterium↗

Determination of VOC source signature of vehicle exhaust in a traffic tunnel.

This study describes the methodology used to obtain the volatile organic compound (VOC) source signature of vehicle exhaust. To accomplish this, C(2)-C(9) VOCs were measured in a traffic tunnel located in Seoul, South Korea. The effect of VOC concentrations from the outside ambient air was considered in the determination of the source signature. To examine the effects of ambient air on VOC concentrations inside the tunnel, the ratio of propane to the total VOC concentrations was compared between the entrance and middle sites in the tunnel. Propane was used as a standard not only because of its insignificant contribution to vehicle exhaust gas, but also the fact that propane is the most abundant VOC in the atmosphere of Seoul. The ratio of propane to the total VOC concentrations was higher at the entrance site than at the middle location by, on average, 60%. This suggests that ambient air affects the inside tunnel air to a greater extent at the entrance site as compared to the middle site. The contribution of ambient air to the air inside the tunnel at the entrance location varied from 30% to 67%, with an average of 55%. This is 1.5 times higher than the value measured at the middle location, which ranged from 20% to 48%, with an average of 36%. This shows that ambient air substantially affects the inside air of the tunnel. Excluding the effects of ambient air on the air inside the tunnel can provide an improved chemical composition for vehicle exhaust using tunnel measurements. We believe that the concentration difference between the two sites within the tunnel provides a more accurate chemical composition of vehicle exhaust as compared to that obtained from a measurement taken at only one point inside the tunnel.

Environmental Monitoring↗

Liquefied petroleum gas (LPG) poisoning: report of two cases and review of the literature.

Two autopsy cases of men who died while connecting a liquefied petroleum gas (LPG) pipe are reported. Their blood concentrations of propane (the main content of LPG) were 0.12 and 3.40 mg/100 g, respectively. The cause of death after exposure of LPG has generally been considered to be asphyxia from hypoxia. The large differences in the blood propane levels found here and reported in the literature, however, suggest that direct toxic effects of propane poisoning may be the cause of death in some cases. Propane concentrations and the cause of death are reviewed and discussed.

Adult↗

A novel gene encoding a 54 kDa polypeptide is essential for butane utilization by Pseudomonas sp. IMT37.

Twenty-three propane- and butane-utilizing bacteria were isolated from soil samples collected from oilfields. Three of them have been identified as Rhodococcus sp. IMT35, Pseudomonas sp. IMT37 and Pseudomonas sp. MT40. SDS-PAGE analysis of the membrane of Rhodococcus sp. IMT35 revealed the presence of at least four polypeptides induced by propane. Polyclonal antibody raised against a 58 kDa polypeptide from Rhodococcus sp. IMT35 specifically detected bacteria which were actively utilizing propane or butane. Immunoscreening of a genomic library in lambdagt11 with this antibody resulted in isolation of a clone containing a 4.9 kb EcoRI genomic DNA fragment. This 4.9 kb DNA fragment was found to hybridize specifically with organisms which could grow on propane or butane. This fragment could therefore be used as a probe for detection of such bacteria. A 2.3 kb fragment having an ORF encoding a polypeptide of 54 kDa was identified by screening a genomic library of Pseudomonas sp. IMT37 with this 4.9 kb EcoRI fragment. The sequence of the ORF (designated orf54) was found to be novel. Primer extension and S1 nuclease mapping showed that transcription of the ORF starts at base 283 and it had sequences upstream similar to that of a Pseudomonas promoter (-12, -24 type). Disruption of the ORF by a kanamycin ('kan') cassette prevented the organism from growing on any alkane but did not affect its ability to utilize the respective alkanols and acids, indicating that alcohol dehydrogenase and subsequent steps in the pathway remained unaltered. The mutants had no detectable level of butane monooxygenase activity. Therefore, the product of this gene plays a crucial role in the first step of the pathway and is an essential component of monooxygenase. The findings imply that this bacterium either employs a common genetic and metabolic route or at least shares the product of this gene for utilization of many alkanes.

Bacterial Proteins↗

The relative efficiency of cryogens used for plunge-cooling biological specimens.

Coolants used for freezing biological specimens were tested for cooling performance in the continuous plunge mode. Results from bare thermocouples showed that ethane cooled faster than propane or a propane:pentane mixture, even when warmed to 25 K above its freezing point. Propane coolants were more efficient than Freon 22 and the slowest cooling occurred in boiling liquid nitrogen. Hydrated gelatin specimens showed similar results with ethane cooling about 33% faster than propane. Epoxy resin specimens cooled faster than hydrated gelatin specimens of similar size. Hydrated and resin specimens cooled over increasing distances as plunge velocity increased. A bare thermocouple, however, cooled over a constant distance when plunged above a critical velocity. This phenomenon may reflect vapour formation and its suppression at high plunge velocities. The rate of cooling in hydrated specimens is shown to have an absolute limit and cannot be modelled by bare thermocouples or resin specimens.

Epoxy Resins↗

Enzymatic activities of rat liver cytosol 10-formyltetrahydrofolate dehydrogenase.

10-Formyltetrahydrofolate dehydrogenase (10-FTH-FDH: EC 1.5.1.6) catalyzes the NADP(+)-dependent oxidation of 10-formyltetrahydrofolate (10-HCO-H4PteGlu) to tetrahydrofolate (H4PteGlu) and CO2 and the NADP(+)-independent hydrolytic cleavage of 10-HCO-H4PteGlu to H4PteGlu and formate. 10-FTHFDH has a 485 amino acid domain at the C-terminus which is 46% identical to aldehyde dehydrogenase (ALDH: EC 1.2.1.3) and contains a conserved active site cysteine (Cys-707). 10-FTHFDH catalyzed NADP(+)-dependent oxidation of propanal and the hydrolysis of p-nitrophenyl acetate (pNPA) in a similar fashion to ALDH. Initial rate studies gave Km values of 46 and 636 microM, respectively, for NADP+ and propanal, while pNPA had a Km of 220 microM. Propanal was able to compete with 10-HCO-H4PteGlu for NADP(+)-dependent oxidation but had no effect on the NADP(+)-independent hydrolase reaction. N-Ethylmaleimide inhibited NADP(+)-dependent 10-HCO-H4PteGlu oxidation but only partially inhibited (65%) hydrolase activity. Disulfiram, a potent inhibitor of cytosolic ALDH, inhibited NADP(+)-dependent propanal oxidation by 10-FTHFDH. We propose that the dehydrogenase reaction of 10-FTHFDH has a mechanism which proceeds through thiohemiacetal and thioester intermediates, similar to that described for aldehyde dehydrogenase. 10-FTHFDH hydrolase activity was dependent on 2-mercaptoethanol and is probably an artifact of the assay system. The N-terminal domain of 10-FTHFDH shows identity to glycinamide ribonucleotide transformylase (EC 2.1.2.2) and contains a putative 10-HCO-H4PteGlu binding site but shows no GAR-TF activity. NADP(+)-dependent oxidation of 10-HCO-H4PteGlu by 10-FTHFDH was inhibited by the folate anti-metabolite, 5,10-dideazatetrahydrofolate, a known GAR-TF inhibitor.

Aldehydes↗

Phenylhydrazine-induced lipid peroxidation of red blood cells in vitro and in vivo: monitoring by the production of volatile hydrocarbons.

Human red blood cells and male Sprague-Dawley rats were treated in vitro and in vivo, respectively, with phenylhydrazine in order to determine whether the release of volatile hydrocarbons can serve as a suitable index for phenylhydrazine-induced red blood cell peroxidation. Lipid peroxidation following phenylhydrazine administration (in vitro experiments: dosage calculated at 0.5-50 mM; in vivo experiments: intraperitoneal injection of 2.8 mg/100 g body wt) was monitored by the release of ethane and pentane measured by gas chromatography. Further hydrocarbons such as ethylene, propane, n-butane, iso-butane and iso-butene were monitored to form a basis of comparison. In vitro haemolysis was also determined during the course of incubation. Red blood cell suspensions yielded more than 15-fold concentrations of propane and more than 2-fold concentrations of iso-butane compared to pentane and ethane yields. Haemoglobin solutions also produced propane and iso-butane in the presence of phenylhydrazine, whereas pentane and ethane were not detectable. Time-course studies revealed that ethane and pentane reached maximum in vitro levels after red blood cell suspensions had been incubated for 2 hr whereas the maximum degree of haemolysis (approximately 60%) was attained between 60 and 90 min following the beginning of phenylhydrazine treatment. The dosage did not affect the final degree of haemolysis. Rats treated with phenylhydrazine exhaled greater concentrations of ethane (6-fold increase) and pentane (2-fold increase) compared to control animals. Exhaled propane showed a 30-fold increase in concentration following drug treatment. Our results suggest that the release of pentane and ethane may be useful in assessing red blood cell lipid peroxidation in the presence of phenylhydrazine in vitro and in vivo.

Alkanes↗