Search PubMed⌕ Search

Biomedical subjects

E Zeiger

Publications and source records attributed to E Zeiger.

At least 127 records · Page 7Linked to original sources
PubMed1986

Mutagenesis, clastogenesis, carcinogenesis: expectations, correlations and relations.

The expectations of the ability of in vitro mutagenesis and clastogenesis assays to unambiguously detect chemical carcinogens have not been fulfilled. The results of this study show a constant error rate in the identification of noncarcinogens with mutagenic potential. Also, different chemical carcinogen classes showed different correlations with mutagenicity. The relationship is further complicated by differential patterns of tumor induction in rodents. Therefore, while in vitro mutagenicity and clastogenicity define important properties of chemicals, additional information is required in order to clearly relate such results to potential carcinogenicity.

Animals↗
PubMed1985

Development of a standard protocol for in vitro cytogenetic testing with Chinese hamster ovary cells: comparison of results for 22 compounds in two laboratories.

A major problem of cytogenetics testing in mammalian cells is lack of agreement of results among laboratories. Our objective was to develop a sensitive in vitro test protocol that was applicable to large-scale chemical screening and yielded comparable results in two laboratories. We used sister chromatid exchange (SCE) and chromosome aberration (CAb) tests in Chinese hamster ovary (CHO) cells. The initial protocol used standard cell densities, medium, batch of rat liver S9 for metabolic activation; positive, negative, and solvent controls; staining and scoring techniques; and fixation times. Treatment without S9 was for 8-12 hr (CAb) or 26 hr (SCE), and with S9 for 2 hr in serum-free medium. Bromodeoxyuridine (BrdUrd) (10 microM) was added to SCE cultures only, 2 hr after addition of the test chemical. Doses were based on the 50% toxicity level in a preliminary test of cell survival 24 hr after treatment. One hundred cells (CAb) or 50 cells (SCE) were scored from each control and from five dose levels. Five clastogens were tested in the first two-laboratory comparison: mitomycin-C, triethylenemelamine, N-methyl-N'-nitro-N-nitrosoguanidine, cyclophosphamide, and benzo(alpha)pyrene. There was quite good agreement between laboratories. Seventeen compounds were then tested "blind" in the two laboratories. As testing proceeded, some discrepancies occurred between the laboratories, and the protocol was modified in attempts to improve the resolution of marginal responses and make dose selection more consistent. The preliminary test for cell survival was omitted. A 10(5) dose range in a half-log series was tested, and cells were scored at the highest dose at which sufficient mitotic cells were obtained, and at the next two lower doses. By delaying fixation times, SCE and CAb were scored at doses that inhibited cell cycle progression. This protocol gave comparable results in the two laboratories in many cases and by testing up to a maximum dose, limited by solubility and/or toxicity, should detect a high proportion of clastogens and SCE inducers.

Animals↗
PubMed1985

Mutagenicity testing of di(2-ethylhexyl)phthalate and related chemicals in Salmonella.

Di(2-ethylhexyl)phthalate and 33 other phthalates, ethylhexanol derivatives, and related chemicals were tested for mutagenicity in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 without metabolic activation and in the presence of rat and hamster liver S-9 metabolic activation systems. No mutagenic activity was seen with any of the chemicals tested.

Animals↗
PubMed1985

The stability of mutagenic chemicals stored in solution.

Using the Salmonella/microsome assay, we have evaluated the stability of mutagenic responses of chemicals stored frozen over a period of 18 months. Each of the standard mutagens was prepared in January 1982, and aliquots were stored at -20 degrees C and at -80 degrees C. Sodium azide (NaN3) was dissolved in water; 4-nitro-o-phenylenediamine (4NOP), 4-nitroquinoline-N-oxide (4NQO), benzo(a)pyrene (B[a]P), and 2-aminoanthracene (2AA) were dissolved in DMSO, all at 100 micrograms/ml. At various times, aliquots were removed, thawed, and tested in parallel with freshly prepared mutagen samples using strain TA100 in a standard plate test and freshly prepared Aroclor 1254-induced rat liver S-9 mix where needed. 4NOP (2-10 micrograms/plate), 4NQO (0.01-0.10 micrograms/plate), B(a)P (0.5-2.5 micrograms/plate), and 2AA (0.25-2.0 micrograms/plate) showed no significant differences between the freshly prepared solutions and the solutions stored at -20 degrees C and -80 degrees C. NaN3 (0.1-0.8 micrograms/plate) did show a statistically significant difference, with the fresh samples giving the lowest mean responses (over all doses) and the -80 degrees C treatment giving the highest. The freezing of mutagen solutions is adaptable to routine use and provides the advantage of reducing the time required to prepare positive control chemicals and reducing the exposure of laboratory personnel to known mutagens.

4-Nitroquinoline-1-oxide↗
PubMed1985

Strategies to reduce the cost of mutagenicity screening with the Salmonella assay.

An extensive Salmonella assay database was analyzed in order to develop strategies to reduce costs of screening chemicals for mutagenicity. This database was obtained from testing 941 samples (representing 799 chemicals), 36% of which were judged mutagenic. Strains TA98, TA100, TA1535, and TA1537 without activation, with rat liver S-9, and with hamster liver S-9, make up the 12 strain/activation combinations considered here. The testing strategies examined consist of two or three stages; a positive result at any stage is regarded as definitive and stops the testing. Sequential testing improves efficiency by eliminating the need for further experimentation once a chemical has been found to be mutagenic. Consequently, costs and effort are reduced. For screening chemicals in the Salmonella assay, it is our recommendation that a sequential testing scheme be adopted whose initial stage consists of TA100.

Animals↗
PubMed1985

Reproducibility of microbial mutagenicity assays: II. Testing of carcinogens and noncarcinogens in Salmonella typhimurium and Escherichia coli.

A total of 63 chemicals were tested for mutagenicity in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538, and Escherichia coli WP2 uvrA in a four-laboratory study. Sixty of the chemicals had been tested for carcinogenicity by the National Cancer Institute or the National Toxicology Program. All chemicals were tested for mutagenicity without metabolic activation and with liver S-9 preparations from uninduced and Aroclor 1254-induced F344 rats, B6C3F1 mice, and Syrian hamsters. The intra- and interlaboratory reproducibility of the Salmonella assay with regard to the overall judgment of mutagenic or nonmutagenic was good. The results in the E coli strain, however, exhibited a high degree of variability between laboratories. With one or two exceptions, the mutagens were detected with S-9 preparations from all three species. The uninduced liver S-9 preparations did not activate any chemicals to mutagens that were not also activated by induced S-9, but some chemicals were detected as mutagens only when induced S-9 was used. A positive mutagenic response in Salmonella was predictive of carcinogenicity 69% of the time; when equivocal carcinogens and borderline mutagens were included, the predictivity increased to 83%. Conversely, 76% of the carcinogens were mutagens. When the equivocal carcinogens were included, the proportion dropped to 75%. Relatively few chemicals (18%) were mutagenic in E coli. Not all the carcinogens induced tumors in both rats and mice, and the species-specific carcinogenicity could not be predicted from the S-9-specific mutagenicity.

Animals↗
PubMed1985

Mutagenicity of pyrene in Salmonella.

Pyrene was tested for mutagenicity in Salmonella typhimurium strains TA97, TA98, TA100 and TA1537. Mutagenicity was seen in all strains when S9 was present.

Animals↗
PubMed1985

Kinetic properties of the blue-light response of stomata.

The stomatal response to blue light was analyzed with gas-exchange techniques in Commelina communis L. leaves by using high-fluence-rate short pulses. Pulses of blue light were given under a background of high-fluence-rate red light, which maintained photosynthesis at near saturation and stomatal conductance at a steady state. A single blue light pulse of 1-100 sec induced an increase in stomatal conductance, which peaked after 15 min and then returned to the initial steady-state level within 50-60 min after the pulse. The response could be repeatedly induced in the same leaf. Red light pulses on a red background did not induce any comparable response. The stomatal response quantified by integrating the conductance increases after pulse application approached saturation with increasing pulse duration (t((1/2)) approximately 9 sec with 250 mumol.m(-2).sec(-1) of blue light). After a saturating pulse, sensitivity to a second pulse was restored slowly. This recovery response, quantified from the conductance increases caused by the two pulses, approached saturation with a t((1/2)) of approximately 9 min. These results were used to test a model in which a molecular component in the sensory transduction process is considered to exist in two interconvertible forms, A and B. If B is the physiologically active form inducing stomatal opening, then A is the inactive form. The A to B conversion is a light-induced reaction and the B to A conversion is a thermal reaction. Rate constants for these reactions were estimated from single- and double-pulse experiments (at a fluence rate of 250 mumol.m(-2).sec(-1), k(1) = 0.075 sec(-1); thermal rate constant k(d) = 0.0014 sec(-1)), allowing the calculation of steady-state concentration of B under continuous irradiation. The calculated values accurately predicted the steady-state stomatal conductances under continuous blue light.

Journal Article↗
PubMed1985

Stomatal Limitation to Carbon Gain in Paphiopedilum sp. (Orchidaceae) and Its Reversal by Blue Light.

Leaves from Paphiopedilum sp. (Orchidaceae) having achlorophyllous stomata, show reduced levels of stomatal conductance when irradiated with red light, as compared with either the related, chlorophyllous genus Phragmipedium or with their response to blue light. These reduced levels of stomatal conductance, and the failure of isolated Paphiopedilum stomata to open under red irradiation indicates that the small stomatal response measured in the intact leaf under red light is indirect.The overall low levels of stomatal conductance observed in Paphiopedilum leaves under most growing conditions and their capacity to increase stomatal conductance in response to blue light suggested that growth and carbon gain in Paphiopedilum could be enhanced in a blue light-enriched environment. To test that hypothesis, plants of Paphiopedilum acmodontum were grown in controlled growth chambers under daylight fluorescent light, with or without blue light supplementation. Total photosynthetic photon flux density was kept constant in both conditions. Blue light enrichment resulted in significantly higher growth rates-of up to 77%-over a 3 to 4 week growing period, with all evidence indicating that the blue light effect was a stomatal response. Manipulations of stomatal properties aimed at long-term carbon gains could have agronomic applications.

Journal Article↗
PubMed1985

Stomatal Responses to CO(2) in Paphiopedilum and Phragmipedium: Role of the Guard Cell Chloroplast.

A role of the guard cell chloroplasts in the CO(2) response of stomata was investigated through a comparison of the leaf gas exchange characteristics of two closely related orchids: Paphiopedilum harrisianum, which lacks guard cell chloroplasts and Phragmipedium longifolium, which has chlorophyllous guard cells. Leaves of both species had an apparent quantum yield for assimilation of about 0.05, with photosynthesis saturating at 0.300 to 0.400 millimoles per square meter per second. CO(2) curves were obtained by measuring steady-state assimilation and stomatal conductance under 0.180 or 0.053 millimoles per square meter per second white light, or darkness, at 0 to 400 microliters per liter ambient CO(2). The response of assimilation to changes in CO(2) was similar in the two species, but the response of conductance was consistently weaker in Paphiopedilum than in Phragmipedium. The data suggest involvement of guard cell chloroplasts in the stomatal response to CO(2) and in the coupling of assimilation and conductance in the intact leaf.

Journal Article↗
PubMed1985

Cyclic and Noncyclic Photophosphorylation in Isolated Guard Cell Chloroplasts from Vicia faba L.

High rates of both cyclic and noncyclic photophosphorylation were measured in chloroplast lamellae isolated from purified guard cell protoplasts from Vicia faba L. Typical rates of light-dependent incorporation of (32)P into ATP were 100 and 190 micromoles ATP per milligram chlorophyll per hour for noncyclic (water to ferricyanide) and cyclic (phenazine methosulfate) photophosphorylation, respectively. These rates were 50 to 80% of those observed with mesophyll chloroplasts. Noncyclic photophosphorylation in guard cell chloroplasts was completely inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea supporting the notion that photophosphorylation is coupled to linear electron flow from photosystem II to photosystem I. Several lines of evidence indicated that contamination by mesophyll chloroplasts cannot account for the observed photophosphorylation rates.A comparison of the photon fluence dependence of noncyclic photophosphorylation in mesophyll and guard cell chloroplasts showed significant differences between the two preparations, with half saturation at 0.04 and 0.08 millimole per square meter per second, respectively.

Journal Article↗
PubMed1984

Suppressive effects of chemicals in mixture on the Salmonella plate test response in the absence of apparent toxicity.

There are chemicals that affect the number of his+ revertant colonies of Salmonella in the plate test at doses that are apparently nontoxic, but may be causing nonlethal, toxic effects. When mixed with mutagens, these chemicals reduce the numbers of his+ revertant colonies on the plate with no accompanying visible toxic effect on the background lawn. Some of these plates are indistinguishable from spontaneous control plates, leading to the possibility that the mutagens under test would be evaluated as nonmutagenic, or that the mutagenic response would be underestimated. The reduction in mutagen-induced revertant colonies in most cases is equivalent to the reduction in spontaneous revertants in the absence of mutagen. A spot test that permits a rapid screen of chemicals for inhibitory effects has been developed; a plate incorporation assay is used to confirm the effect. Toxic effects can be seen in the background lawns of plates examined at magnifications of 100X or greater.

Animals↗
PubMed1984

Mutagenicity testing of agent orange components and related chemicals.

Components of the herbicide Agent Orange--2,4-dichlorophenoxyacetic acid (2,4,-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and their esters, and the contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)--and related chemicals were tested for mutagenicity using Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537. No mutagenic activity was observed for any of the chemicals tested.

2,4,5-Trichlorophenoxyacetic Acid↗
PubMed1984

Sources of variability in Ames Salmonella typhimurium tester strains: analysis of the International Collaborative Study on 'genetic drift'.

Data from 38 laboratories using 5 strains of Salmonella typhimurium (TA98, TA100, TA1535, TA1537, and TA1538) were analyzed to determine sources and magnitudes of test data variability. Each laboratory tested the mutagenicity of 4-nitroquinoline-N-oxide by the same protocol, using both its in-house cultures and a set of reference cultures provided to all laboratories. It was found that neither plate-to-plate nor day-to-day variability within a laboratory differed substantially between the in-house and reference cultures for any strain; this indicated no difference in the laboratories' handling of the two cultures. Not surprisingly, on average, plate-to-plate variability was substantially smaller than day-to-day variability within a laboratory, which, in turn, was substantially smaller than inter-laboratory variability. The solvent DMSO was found to have a small (6-7%) but statistically significant depressive effect on the spontaneous mutant frequency for the two plasmid-containing strains, TA98 and TA100, but not for the other three strains. When the mean value and variance of all laboratories for the in-house culture were compared with the corresponding reference culture values for each dose and strain, no major differences were seen. Any increase in mean or variance in the distribution of laboratory means in one of the two cultures could be ascribed largely to a small number of laboratories. Laboratories that reported 'high' or 'low' levels of spontaneous or induced revertants per plate tended to deviate in the same direction for most strains and for both in-house and reference cultures. If 'genetic drift' contributed to the inter-laboratory variability in this collaborative study, it was a minor component that went undetected in our analyses.

4-Nitroquinoline-1-oxide↗
PubMed1983

The intrasanguineous host mediated assay procedure using Saccharomyces cerevisiae: comparison with two other metabolic activation systems.

3 metabolic activation systems--organ homogenates, perfused liver, and the intrasanguineous host mediated assay (IHMA)--were compared in their abilities to activate demethylnitrosamine (DMN) and induce gene conversion in Saccharomyces cerevisiae D4. Both rats and mice were used for the organ homogenates and IHMA studies. All activation systems were able to activate DMN; where the different organs were compared, liver was more active than kidney, followed by lung. The IHMA was the most sensitive of the systems examined.

Animals↗
PubMed1983

Specificity of rat liver cytochrome P-450 isozymes in the mutagenic activation of benzo[a]pyrene, aromatic amines and aflatoxin B1.

The ability of three purified forms of rat liver cytochrome P-450 to metabolically activate benzo[a]pyrene, trans-benzo-[a]pyrene-7,8-dihydrodiol, 2-aminofluorene, aflatoxin B1, dimethylnitrosamine, and a pyrolysis product of tryptophan(3-amino-1-methyl-5H-pyrido(4,3-b)indole) (Trp-P-2) to mutagenic products was examined using Salmonella typhimurium strains TA98 and G46 in a reconstituted monooxygenase system. The isozymes examined were cytochrome P-450-PB (the major phenobarbital inducible form), and the two major 3-MC inducible forms (cytochromes P-448(52) and P-448(55)). Cytochromes P-448(52) and P-448(55) preferentially metabolize 2-aminofluorene and Trp-P-2 to mutagenic products. However, only cytochrome P-448(55) metabolizes benzo[a]pyrene and its 7,8-dihydrodiol derivative to mutagenic products. Both cytochrome P-448(52) and P-448(55) metabolize aflatoxin B1 to mutagenic products at a much faster rate than cytochrome P-450-PB. Dimethylnitrosamine was not activated by any of the isozymes tested.

Aflatoxin B1↗