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PubMed2002

Analysis of surface properties of fixed and live cells using derivatized agarose beads.

A novel assay has been developed for the histochemical characterization of surface properties of cells based on their adhesion to agarose beads derivatized with more than 100 types of molecules, including sugars, lectins and other proteins, and amino acids. The assay simply involves mixing small quantities of washed cells and beads in droplets on glass microscope slides and determining to which beads various cell types adhere. Distilled water was found to be the best medium for this assay because added ions or molecules in other media inhibit adhesion in some cases. Many cells, however, cannot tolerate distilled water. Here we show that cells fixed with either of two fixatives (1% formaldehyde or Prefer fixative) displayed similar bead-binding properties as did live cells. Specificity of cell-bead binding was tested by including specific free molecules in the test suspensions in hapten-type inhibition experiments. If a hapten compound inhibited live-cell adhesion to a specific bead, it also inhibited fixed-cell adhesion to a specific bead. The results of these experiments suggest that fixed cells display authentic surface properties, opening the door for the use of this assay with many cell types that cannot tolerate distilled water.

Cell Membrane↗

PubMedDate not supplied

Breath ammonia depletion and its relevance to acidic aerosol exposure studies.

It is thought that gaseous ammonia in breath neutralizes acidic air pollution and thereby potentially mitigates the pulmonary effects of pollution. The efficacy of breath ammonia depletion methods reported in recent acid aerosol exposure-health response studies was investigated. Fourteen subjects (21 to 54 y of age) performed one or more of the following hygiene maneuvers: (a) acidic oral rinse (pH 2.5); (b) tooth brushing, followed by acidic oral rinse; (c) tooth brushing, followed by distilled water rinse; and (d) distilled water rinse. Initial ammonia levels ranged from 120 to 1,280 ppb (147-1,570 micrograms/m3). Acidic rinsing resulted in an immediate 90% reduction in exhaled ammonia in all subjects, and a return to 50% of baseline levels occurred within 1 h. Depletion that resulted from tooth brushing or distilled water alone was not significant. It was concluded that acidic oral rinsing is an effective method of reducing airway ammonia, but repeated oral rinsing may be required to maintain consistent, low-breath-ammonia conditions during acid aerosol exposure studies.

Adult↗

PubMed2001

Migration of 4-nonylphenol from polyvinyl chloride food packaging films into food simulants and foods.

Migration of 4-nonylphenol (NP) from polyvinyl chloride (PVC) films for food packaging into food simulants and foods has been studied in domestic applications such as wrapping of food and reheating in a microwave oven. The migration of NP from the PVC films was determined by high-performance liquid chromatography with electrochemical coulometric-array detection (LC/ED). Twelve PVC films intended for commercial use and ten for domestic applications (total: 22 samples) were analysed. Some of the PVC films (two home-use and ten retail-use) contained NP at concentrations of between 500 and 3300 microg/g. Migration of NP from the films was influenced by the test conditions (n-heptane at 25 degrees C for 60 min, distilled water at 60 degrees C for 30 min and 4% acetic acid at 60 degrees C for 30 min). The amount of NP migrating from the PVC films into n-heptane (0.33-1.6 microg/cm2) was higher than the amount migrating into distilled water or 4% acetic acid (up to 9.7 ng/cm2) for the 11 films in which NP was detected. Up to 0.23% of the NP migrated into distilled water and 4% acetic acid and up to 62.5% into n-heptane. In addition, we investigated NP migration into cooked rice samples wrapped in PVC film. Using spiked samples the method gave an average recovery of 83.7% (n = 5) with a standard deviation of 2.5%. Migration of NP ranged from not detectable (< 1.0 ng/g) to 410.0 ng/g by reheating samples in a microwave oven for 1 min and from not detectable to 76.5 ng/g by keeping samples at room temperature for 30 min.

Chromatography, High Pressure Liquid↗

PubMed2000

The determination of total SO2 in grape juice. A comparison among five methods.

The EC official method of total SO2 analysis in grape juice was modified in 1990. The main improvements concerned the amount and concentration of H3PO4 used during the distillation to recover the combined SO2 and the standardization of the distillation time at 15 min. This comparative study evaluated the total SO2 level of 12 grape juices determined by five methods, including distillation, iodimetric and enzymatic-based methods. Attention was focused on the total SO2 legal limit of 10 mg/l fixed in Europe for grape juice. Analysis of variance disclosed a significant difference among the total SO2 content in grape juices determined by five methods. Each analytical method showed limits in relation of their ability to release the combined SO2. In particular, the SO2 bonded to phenolic compounds is partially released at low pH in the acidified juice leading to higher results.

Analysis of Variance↗

PubMed2003

Migration of 2-butoxyethyl acetate from polycarbonate infant feeding bottles.

An enforcement campaign was carried out to assess the migration of 2-butoxyethyl acetate (2-BEA) from polycarbonate infant feeding bottles intended for repeated use. Migration was measured by three successive migration tests into two of the European Union official food simulants: distilled water and 3% acetic acid testing at 40 degrees C for 10 days. The Danish Veterinary and Food Administration (DVFA) has assessed that a migration above 0.33 mg for 2-BEA and a group of eight related substances kg(-1) foodstuff from plastics articles used exclusively for infants is unacceptable. Migration of 2-BEA was found from eight of 12 bottles. However, migration above the target value of 0.33 mg kg(-1) was not observed in the third decisive test from any of the 12 different brands of polycarbonate feeding bottles. A migration of between 0.05 and 0.26 mg kg(-1) from seven of 12 bottles was measured to 3% acetic acid in the third test, whereas no migration to distilled water was observed in the third test. The average recovery of 2-BEA after the 10-day exposure at the target value of 0.33 mg kg(-1) was 77% into distilled water and 36% into 3% acetic acid. The limited recovery was understandable as 2-BEA was partly hydrolysed in the aqueous food simulants and 2-butoxyethanol, a hydrolysis product and one of the related substances was identified. Quantification was carried out using gas chromatography after liquid/liquid extraction of the food simulant.

Acetates↗

PubMed2005

Migration of styrene monomer, dimers and trimers from polystyrene to food simulants.

Migration experiments with polystyrene were performed in two-sided contact with n-heptane and distilled water as the food simulants at temperatures of 10, 24 and 40, and 40, 60 and 90 degrees C, respectively. The surface/volume ratios in the migration cell were set at 8.04 and 10.05 dm2 l(-1) for n-heptane and distilled water, respectively. Styrene monomer, styrene dimers and styrene trimers migrating to the food simulants were determined by GC-FID analysis. Heptane fully extracted the styrene monomer and the oligomers from the polystyrene sheet, whereas in the distilled water only the migrated styrene trimers could be detected. To determine the apparent diffusion coefficient, the migration process was analysed based on Fick's law. The higher the molecular weight of the oligomers, the more significant the reduction in the diffusion coefficient. Higher molecular weight oligomers also had lower activation energy of diffusion when the temperature dependence of diffusivity was analysed by the Arrhenius equation. The diffusion coefficient of the trimers was much higher for heptane contact than for water. The activation energy of the diffusion of trimers for water contact was higher than that for heptane.

Chromatography, Gas↗

PubMedDate not supplied

Food-contact epoxy resin: co-variation between migration and degree of cross-linking.

In order to predict the behaviour towards foodstuffs of an epoxy resin composed of bisphenol A diglycidyl ether (BADGE), 4,4'-methylenedianiline (MDA) and additives (plasticizers: dibutylphthalate (DBP), dioctylphthalate (DOP); accelerator: salicylic acid; inorganic fillers), a co-variation was established between the parameters evaluating the degree of cross-linking of the three-dimensional network and the migration of constituent molecules into various food simulants (distilled water, distilled water/ethanol/acetic acid, distilled water/ethanol). Varied degrees of cross-linking were obtained by subjecting the resin to different curing temperatures: respectively, 5 degrees C, 20 degrees C, 50 degrees C and 90 degrees C for 7 days. Irrespective of the food stimulant tested, specific migrations (DBP, DOP, salicylic acid, primary aromatic amines) diminished greatly as the curing temperature increased. At the same time, the degree of cross-linking increased with curing temperature, as indicated by the increase in glass transition temperature, the decrease in residual reaction exotherms and increased stability of the rubber storage modulus E'rub (increase in cross-link nodes), the fall in relaxation enthalpies (reduction in physical ageing) and the decreased amplitude of the loss-factor, tan delta (reduction in chain mobility). Maximum cross-linking was obtained in the resin cured at 90 degrees C (temperature above Tg infinity). In contrast to the degree of cross-linking, evaporation contributed little to the reduction of migration due to the elevation of curing temperature.

Chemical Phenomena↗

PubMed1998

Food-contact epoxy resin: co-variation between migration and degree of cross-linking. Part II.

The study of epoxy resin composed of bisphenol A diglycidylether (BADGE), bisphenol F diglycidylether (BFDGE) (base), and primary aliphatic polyamines (hardener), has confirmed the interest of measuring certain physical parameters in order to evaluate the density of cross-linking of the network and thus predict the risks of resin molecules migrating into foodstuffs. This suggestion had been made in a preceding study on an epoxy resin composed of bisphenol A diglycidylether (BADGE) and primary aromatic polyamines. Samples with different densities of cross-linking, obtained by subjecting the resin to different curing temperatures (5, 20, 50 and 90 degrees C) for 7 days, were studied. The density of cross-linking increased with curing temperature, as indicated by the increase in glass transition temperature, the increased stability of the rubber storage modulus E'rub (increase in cross-link nodes), the fall in relaxation enthalpies (reduction in physical ageing) and the decreased amplitude of the loss-factor tan delta (reduction in chain mobility). Maximum cross-linking was obtained in the resin cured at 90 degrees C (temperature above Tg infinity). Concurrently, tests of migration into different liquid food simultants (distilled water, distilled water/ethanol/acetic acid, distilled water/ethanol) revealed a considerable reduction in specific migrations of BADGE and BFDGE, and of unidentified peaks.

Chromatography, Gas↗

PubMed2003

Microbial and copper adsorption by smectitic clay--an experimental study.

The objective of this study was to quantify copper-, bacteria- and bacteriophage-binding capacities of natural clay with the aim of predicting the adsorption of heavy metals, human pathogenic bacteria and viruses by a clayey landfill liner. X-ray diffraction analysis of six natural clays showed that the dominant phase in all deposits consists of smectites together with illite, kaolin and, sometimes, palygorskite and sepiolite. The specific surface areas of different clay substrates were very high ranging from 293 to 351 m2 g(-1), and indicating a high proportion of phyllosilicates, consisting especially of smectites. The physico-chemical identification of separated smectites showed a high potential adsorbent character indicative of a large industrial use. The Kb12 smectite substrate chosen arbitrarily among six separated substrates, appeared as an excellent copper adsorbent. Copper was adsorbed to clay in a proportion ranging from 94.6 to 96.0% with an average of 95.1% and its adsorption occurred rapidly in less than 30 min. Organic contents of the clay substrate, evaluated as 17% of dry mass, may contribute and enhance copper adsorption. Different elution protocols using distilled water, 2 and/or 5% nitric acid revealed that while nitric acid resulted in the removal of more than 59% of the metal at the lower concentration and its complete depletion with a further elution at the higher concentration, distilled water alone was unable to remove more than 1% of adsorbed copper. This finding suggested that copper ions form high-energy bonds with layer-silicate surfaces. Interestingly, the use of a regenerated substrate as copper adsorbent subsequent to abundant washings of the used substrate consecutively, with 0.1 N HNO3 and distilled water, reduced copper adsorption by approximately 14%, suggesting a slight disturbance of clay initial structure. Batch adsorption experiments with phage T7 and raw clay Kb12 showed that the tested clay substrate appeared as a relatively moderate phage adsorbent since the quantity of adsorbed phage averaged 98.2 +/- 0.88% (2 log10 retention) as measured by infectivity for Escherichia coli ATCC 11303. As shown by two types of separating procedures, natural sedimentation and a low speed centrifugation, bacteriophage particles were bound essentially to fine and not to relatively coarse particles of the clay suspension. The retention capacity of purified clay Kb12 appeared low, with average values lower than 60 and 50%, for Pseudomonas aeruginosa ATCC 15442 and Bacillus cereus ATCC 1135, respectively. A significant increase of retention, in the order of 30%, was found for both bacteria when the mixture clay-bacteria was incubated at laboratory temperature for 6 hours.

Adsorption↗

PubMed1986

Biological availability of nickel arsenides: cellular response to soluble Ni5As2.

Particulate Ni5As2 has been shown to be highly cytotoxic and carcinogenic. By measuring the solubility of Ni5As2 particles in a variety of aqueous solutions, we have determined that particulate Ni5As2 that might be produced during oil-shale retorting could be mobilized to the environment and made available to the cells of living organisms, including humans. Ni5As2 was five times more soluble in ground water taken from aquifers surrounding a major oil-shale source in Colorado, U.S.A., than in distilled water. It was also two times more soluble in oil-shale product water from an above-ground retort than in distilled water. Thus, it is possible that Ni5As2 could be solubilized and mobilized to the environment by the flooding of abandoned in situ retorts with ground water or by the disposal of oil-shale product water by spraying it on spent shale beds. Particulate Ni5As2 was found to be 12 times more soluble in culture growth medium than in distilled water, and much more soluble in solutions of amino acids, inorganic salts, organic constituents of culture medium, and 15% calf serum. These observations suggest Ni5As2 particles in airborne dust would be dissolved when they came in contact with the biological fluids of the lung and gastrointestinal tract. The availability to cells of the soluble products of Ni5As2 was demonstrated by measuring its effects on cell proliferation. As little as 1 ppm soluble Ni5As2 retarded Chinese hamster (CHO) cell proliferation in culture, and 4 ppm resulted in cell death. Flow cytometry measurements indicated there was a preferential cytotoxic effect on S-phase cells. Despite this, many cells survived to form colonies, causing concern that Ni5As2 might cause genetic damage that could be passed on to future cell generations. This did not appear to be the case, however, for no mutations could be detected at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus in cells that survived the cytotoxic effects. This suggests that Ni5As2 carcinogenesis might be caused by epigenetic rather than mutagenic mechanisms.

Animals↗

PubMedDate not supplied

Effect of acipimox on plasma lipids and glucose/insulin in pregnant rats.

To determine how a reduction in maternal hypertriglyceridemia during late pregnancy may affect glucose/insulin relationships, pregnant and virgin rats were orally treated with acipimox, a potent antilipolytic agent. In 20-day pregnant rats receiving 80 mg of acipimox, plasma triglycerides (TG), free fatty acids (FFA), and glycerol decreased more than in virgin rats shortly after the drug (up to 7 hours), when compared with animals treated with distilled water, whereas plasma glucose level was unaffected by the treatment in either group of rats. When acipimox was given every 12 hours from day 17 to day 20 of pregnancy, plasma TG, FFA, and glycerol levels progressively increased, whereas they either decreased or did not change in virgin rats receiving the same treatment, with no effect in plasma glucose levels in either group. Fetal body weight was lower than in controls in 20-day pregnant rats that received acipimox for 3 days. On day 20 of pregnancy, 3 hours after receiving acipimox or distilled water, rats received a 2 g glucose/kg oral load and it was found that the change in plasma glucose was similar in both groups, whereas the increase in plasma insulin was greater in pregnant rats treated with acipimox. However, no difference was found in either variable after the oral glucose load in virgin rats receiving acipimox or distilled water. No differences in plasma glucose levels were found after intravenous (i.v.) administration of insulin in pregnant rats treated or not treated with acipimox. In conclusion, present results show that administration of acipimox during the last days of gestation inhibited lipolysis and decreased fetal weight. Over a short period of time, in pregnant rats, reductions of plasma FFA and TG after acipimox treatment improved the glucose-induced insulin release, but did not seem to have any effect in peripheral insulin resistance.

Animals↗

PubMed1951

The mechanism of virus attachment to host cells. I. The role of ions in the primary reaction.

T1 virus does not attach to its host cell, E. coli B, in distilled water. By the proper addition of salts the rate of attachment can be adjusted to any desired value up to the maximum limit set by the diffusion rate of the virus. Salts of Ca(++), Mg(++), Ba(++), and Mn(++) bring about a reaction rate representing 100 per cent collision efficiency in a concentration of 5 x 10(-4)M. Both greater and smaller concentrations depress the attachment velocity. Salts of Na(+), K(+), NH(4) (+), and Li(+) display a similar pattern but require a tenfold greater concentration than that of the previous group to produce the same effect. Moreover, the maximum velocity attainable in solutions containing only monovalent cations is only half that achieved by the divalent salts. The trivalent cations Al(3+), Cr(3+), Fe(3+) permanently inactivate the virus. Activation by Mg(++) of an inert mixture of virus and host cells in distilled water is so rapid as to be beyond the limit of the resolving time of the experimental procedure, which is 20 seconds. The temperature dependence curve of virus-cell adsorption exhibits a maximum at 37 degrees C. and falls to a value representing approximately 3 per cent collision efficiency at 1 degrees C. Identical curves are obtained in nutrient broth and in 10(-)M MgCl(2) solution. Bacteriophage can be quantitatively adsorbed on to glass filters. In a study of several viruses this attachment reaction was found to require the same cofactors-both organic, like ltryptophane, as well as inorganic-which each specific virus required for its attachment to its host cell. The suggestion is made that the attachment of viruses to these filters is a useful model for their attachment to host cells. Virus attachment to glass filters is reversible. Such adsorbed virus can be recovered almost quantitatively by washing the filter with a solution in which the attachment reaction does not occur. Virus attachment to host cells is similarly reversible at least in its primary step. Distilled water at 0 degrees C. can produce almost complete liberation of T2 virus from host cells infected in 0.02 M NaCl solution. Two significant differences between the behavior of glass filters and host cells toward T1 virus are: (a) an excess ion concentration fails to inhibit virus attachment to the glass as it does to the host cell; and (b) no decrease in efficiency of attachment to glass occurs at low temperatures. These facts suggest that the inhibiting action on the infective process of excess cations and low temperatures involves chemical groupings on the cell surface, rather than on the virus. There is no detectable attachment whatever of T1 virus to E. coli cells specifically resistant to it, though still susceptible to other viruses. This experiment indicates that the ion-controlled attachment forces here considered are involved in the host-virus specificity. This conclusion is strengthened by the fact that several different viruses with different host-cell specificities have different ionic requirements for cell attachment. All of these observations lend themselves to explanation by a mechanism which pictures an initial addition reaction of cations to specific sites on the surface of the virus in particular, and possibly also of the host cell. Two complementary electrostatic configurations are so produced which can unite in a reaction with a high biological specificity, which yet exhibits 100 per cent collision efficiency. An excess of ions may cover up some of the attachment sites and so inhibit the reaction. By this picture the specificity of virus-cell invasion depends upon the binding energies of sites on both bodies for various ions, and the distribution of these sites over the two surfaces. Possible relationships of such a process to other biological systems are discussed.

Escherichia coli↗

PubMed2004

Three-dimensional visualization and thickness estimation of aortic valve cusps using high-frequency ultrasound.

High-frequency ultrasound techniques are introduced for three-dimensional imaging and thickness estimation of fresh heart valve cusps. Images of porcine aortic valve specimens were acquired within a 12 x 8 x 8 mm3 volume using a VisualSonics VS40 micro-imaging system operating at a 40 MHz centre frequency. Two image volumes were obtained from each of six left coronary cusps. One volume was acquired with the specimen submerged in distilled water and the second volume was acquired through either Hanks physiologic solution or coronary perfusion solution (CPS). The fibrosa, spongiosa and ventricularis were most readily distinguished when the specimen was imaged in distilled water. Colour thickness maps were computed from B-mode image data, and the mean and standard deviations of the thickness were determined for each cusp. In 11 of 12 trials, the image analysis algorithm yielded valid thickness estimates over greater than 98% of the region examined. Mean thickness estimates obtained with specimens submerged in Hanks solution or CPS ranged from 0.66 to 1.03 mm, and submersion in distilled water increased the mean thickness by 20-40%. This observation suggests that the cusps osmotically absorbed water. Information provided by high-frequency ultrasound is expected be valuable for characterizing the morphological properties of heart valves.

Animals↗

PubMed2003

A new in vitro assay of cholesterol adsorption by food and microbial polysaccharides.

The adsorption of total cholesterol by polysaccharides was measured in vitro by enzymatic reactions, including the polysaccharide precipitation procedure. Total cholesterol adsorption capacities, in a mixture of polysaccharide and total cholesterol, were compared for apple pectin, gelrite gellan gum, xanthan gum, high-methoxyl pectin, citrus pectin, high-viscous alginate, low-viscous alginate, dextran, and zooglan. Acidic polysaccharides such as pectins, alginate, and xanthan gum at concentrations of 0.1% (wt/vol) were able to adsorb over 90% of the total cholesterol when dissolved in distilled water, sodium acetate buffer (pH 4.6), or sodium phosphate buffer (pH 7.0). However, total cholesterol adsorptions by gellan and zooglan were dependent upon the salt concentration and pH value, which decreased cholesterol adsorption in the following order by degree: distilled water, acidic pH, and alkaline pH. In particular, total cholesterol adsorption of zooglan was greatly decreased by the addition of sodium chloride. With 0.1% (wt/vol) polysaccharide dissolved in distilled water, the adsorption capacities of alginate, pectins, gellan gum, xanthan gum, and zooglan were 2.9, 2.88, 2.5, 2.9, and 2.4 mg/dL, respectively. However, 0.2% of zooglan was able to completely adsorb the cholesterol (3 mg/dL), whereas dextran did not adsorb cholesterol at all, producing no precipitate with hexadecyltrimethylammonium bromide.

Adsorption↗

PubMed1997

Sensitivity of the cough reflex in young and elderly subjects.

OBJECTIVE: To compare the sensitivity of the cough reflex--which is said to be normal in elderly people--in elderly and young subjects. SUBJECTS: 20 elderly (14 female) subjects, mean (SEM) age 83 (1) years and 20 young (nine female) subjects, mean (SEM) age 27 (1) years, who were all non-smokers. None of the subjects was taking antitussive drugs and none suffered from clinically evident lung, cardiac or neurological disease. Five elderly subjects were unable to perform adequate spirometry and were excluded from analysis. DESIGN AND OUTCOME MEASURES: Each subject inhaled 10 ml of nebulized distilled water and isotonic saline (as placebo) for 30 s, 10 min apart in a randomized double-blind crossover fashion. The cough frequency induced with each treatment was recorded on a click counter. RESULTS: Cough frequency on inhaling distilled water was significantly lower in the elderly group than in the younger group, with a difference of 9.53 (95% confidence intervals: 3.63, 15.4; P < 0.001). None of the subjects coughed when inhaling placebo solution, resulting in significant differences in cough frequencies between distilled water and placebo of 5.87 (2.82, 8.92; P < 0.05) for the elderly group and 15.4 (11.0, 19.8; P < 0.0005) for the younger group. CONCLUSIONS: The sensitivity of the cough reflex appears to be significantly reduced in elderly subjects. This may increase the risk of aspiration and bronchopulmonary infection in old age, even in the absence of respiratory disease.

Adult↗

PubMed1992

Influence of fruit and vegetable juices on the endogenous formation of N-nitrosoproline and N-nitrosothiazolidine-4-carboxylic acid in humans on controlled diets.

Water, green pepper, pineapple, tomato, strawberry, carrot, and celery juices were made 46 mg/100 ml in ascorbic acid by the addition of distilled water or ascorbate. The ability of each juice to inhibit endogenous formation of N-nitrosoamino acids (NAA) in humans was determined in controlled experiments. Sixteen men consumed a standard diet low in nitrate and ascorbic acid for 18 consecutive days. Nitrate (5.24 mmol) and L-proline (4.35 mmol) were given orally on days 3, 5, 7, 9, 11, 13, 15, 17 and 18. On days 3 and 18, L-proline was immediately followed by 100 ml distilled water (positive control). On days 5, 7, 9, 11, 13, 15 and 17, L-proline was immediately followed by 100 ml juice or 46 mg ascorbate in 100 ml distilled water (treatment). Only diet was given in between dosing days to ensure baseline levels of NAA excretion. Urine was collected for 24 h following treatments and analyzed for NAA. Green pepper, pineapple, tomato, strawberry and carrot treatments significantly inhibited N-nitrosoproline (NPRO) formation relative to the positive control. Also, green pepper, pineapple and tomato juices significantly inhibited NPRO formation relative to ascorbic acid alone. Green pepper significantly inhibited N-nitrosothiazolidine-carboxylic acid formation relative to ascorbic acid alone. These data demonstrate that green pepper, tomato, pineapple, strawberry and carrot juice have greater ability to inhibit endogenous nitrosation than would be expected based solely on their ascorbate content.

Adult↗

PubMed1983

Ornithine decarboxylase induction by chemically complex liquids from two solvent refined coal processes.

Studies with a variety of chemically purified substances have suggested that induction of the enzyme ornithine decarboxylase (ODC) in mouse epidermal cells may be a reliable indicator of neoplastic transformation. In an effort to extend these observations on ODC to chemically complex materials, we examined ODC induction by carcinogenic and non-carcinogenic mixtures and compared these results with tumorigenicity data for these materials. For these studies several boiling range fractions and several solvent-derived subfractions from two solvent-refined coal processes (SRC-I and SRC-II) were evaluated for their ability to induce ODC. Single applications of heavy distillate (HD), the SRC-II high-boiling fraction and a potent mouse skin carcinogen, produced ODC induction kinetics which were similar to that for 12-O-tetradecanoylphorbol-13-acetate (TPA). Both HD and TPA stimulated maximal ODC activity 3-5 h after application, with epidermal ODC levels returning to basal levels within 12 h. The magnitude of ODC induction after multiple applications of HD was not as great as that observed for TPA. Single skin applications of TPA and HD also transiently elevated hepatic ODC levels 27- and 7-fold, respectively; however, liver ODC activity did not increase following multiple applications of either chemical. Further, ODC induction by HD was also dose-dependent. Relative to controls, single applications of HD and process solvent (boiling range greater than 250 degrees C) elevated ODC levels 145- to 205-fold, light distillate and light oil (boiling range less than 180 degrees C) increased ODC levels 23- to 32-fold, and middle distillate and wash solvent (boiling range 180-250 degrees C) stimulated less than 2- to 8-fold increases in ODC. Single applications of three solvent-derived subfractions of HD, which are complete carcinogens, induced 3- to 7-fold ODC elevations over background levels; multiple applications of two of these subfractions elevated ODC levels 10- to 22-fold. Of the complex mixtures evaluated during this study, all complete carcinogens induced ODC; however, the magnitude and temporal pattern of induction varied with the material tested.

Animals↗

PubMed2002

Semiquantitative determination of off-notes in mint oils by solid-phase microextraction.

Mint essential oils are produced by the steam distillation of dried or partially dried harvested plant material. In the United States, harvesting is done mechanically so that any weeds found in the field are concomitantly harvested. Steam distillation of contaminated plant material leads to off-notes in the oil, which are currently determined by a sensory panel. Furthermore, nonoptimized distillation conditions can lead to the thermal degradation of carbohydrates and proteins resulting also in the formation of very volatile off top-notes. As a result, the use of a nonequilibrated solid-phase microextraction (SPME) procedure to determine the off-notes is evaluated. The results of this evaluation include a combination of semiquantitative data, odor threshold data, and mathematical data manipulation to ascertain the capabilities of a SPME approach. The results are correlated with sensory panel data to yield a relatively rapid analytical methodology that can be used either in place of or in support of sensory analyses. The main advantage of the technique described is to provide some semiquantitative data in support of the odor-panel screening of mint oils for off-notes. Based on the data presented in this report, it is believed that this has been successfully demonstrated.

Gas Chromatography-Mass Spectrometry↗