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D M Shankel

Publications and source records attributed to D M Shankel.

36 records · Page 2Linked to original sources

Antimutagens against spontaneous and induced reversion of a lacZ frameshift mutation in E. coli K-12 strain ND-160.

Isopropyl-beta-D-thiogalactoside (IPTG), at 0.5 or 1.0 mM, is shown to reduce spontaneous, and to virtually abolish caffeine- or quinacrine-induced reversions of the lacZ frameshift mutation in E. coli ND160. Guanosine at 200 micrograms/ml and Co2+ at 15 micrograms/ml had an erratic partial antimutagenic effect on spontaneous lac+ reversions. All 3 agents reduce caffeine-induced (500 micrograms/ml) mutagenesis. Spermine (250 micrograms/ml) also reduces quinacrine-induced Lac+ reversion frequencies in this system.

Caffeine↗

Effects of the antimutagen cinnamaldehyde on reversion and survival of selected Salmonella tester strains.

The antimutagenic activity of trans-cinnamaldehyde (C6H5CH = CHCHO) on chemically induced mutagenesis has been shown in E. coli. Using the Ames Salmonella typhimurium tester strains TA1535 (hisG46 uvrB rfa) and TA100 (TA1535/pKM101), the effects of cinnamaldehyde on spontaneous reversions and reversions induced by 4-nitroquinoline-N-oxide(4NQO) and ethyl methanesulfonate (EMS) have been examined. To observe the effect of cinnamaldehyde in the absence of functional muc genes, a third strain, TA1535/pGW201 (pKM101 muc140: :Tn5) was included in the testing. Modifications of the standard Ames test procedures and direct-plating techniques were employed to study the "antimutagenic" response exerted by cinnamaldehyde. In all strains tested, concentrations of cinnamaldehyde up to 25 micrograms/ml slightly decreased the number of spontaneous reversions and induced reversions were more markedly reduced. The decreases in the numbers of 4NQO-induced revertants were greater than those decreases which occurred for EMS-induced reversions. There was no effect on viability in 1% (v/v) nutrient broth supplemented minimal medium containing 5-25 micrograms/ml of cinnamaldehyde. Cinnamaldehyde did not display any mucAB dependent or independent specificity against the mutagens used. On minimal medium supplemented with histidine and biotin, concentrations of cinnamaldehyde above 10 micrograms/ml were lethal for the strains tested. When the test medium was supplemented with 1-5% (v/v) liquid nutrient broth, viability was not affected at concentrations up to 25 micrograms/ml. For both TA100 and TA1535 the presence of 20 micrograms/ml of cinnamaldehyde in 1% (v/v) liquid nutrient broth-supplemented minimal glucose broth extended the lag phase for 2-4 h with no effect on survival. Depending on the test procedure employed, decreases in numbers of revertants may reflect lethality rather than antimutagenesis. When used to test for antimutagenesis rather than mutagenesis, modifications of the standard Ames test procedure may mimic an antimutagenic response due to a decrease in the total number of revertants seen even though enough cells survive to produce a background lawn.

Acrolein↗

Effects of antimutagens on prophage induction in E coli lysogenic for lambda.

The effects of caffeine, acriflavine, spermine, quinacrine, and adenosine on spontaneous and UV-induced prophage induction in repair-proficient and -deficient strains of lysogenic Escherichia coli were examined. Caffeine caused a coinducing effect on UV induction in KMBL 160 (repair proficient) and KMBL 163 (uvrA), an anti-inducing effect in KMBL 161 (uvrE), and had no detectable effect in KMBL 164 (uvrC). Acriflavine increased UV induction in KMBL 160 and decreased it in all three repair-deficient mutant strains. Spontaneous induction was reduced by acriflavine in all four strains. Spermine caused a decrease in UV induction in KMBL 160 and KMBL 164, while it had limited effect in KMBL 161 and KMBL 163. For spontaneous induction spermine caused an increase in KMBL 160 and KMBL 163, a decrease in KMBL 164 and no detectable effect in KMBL 161. Quinacrine had no significant effect on spontaneous induction in KMBL 160 and KMBL 164, but caused a decrease in spontaneous induction in KMBL 161 and KMBL 163. Quinacrine produced no effect on UV induction in KMBL 160; however, it decreased UV induction in the three repair-deficient strains to some extent. A significant increase in spontaneous induction in KMBL 160 was caused by adenosine, while it had little effect in the repair-deficient strains. UV induction was not affected by adenosine.

Acriflavine↗

Caffeine as a comutagen for ethylmethanesulfonate in strains of phage T4.

The mutation frequency of DNA polymerase mutants of phage T4 treated with ethyl methanesulfonate (EMS) then incubated in the presence and absence of caffeine was studied using an rII reversion system. The DNA polymerase mutation is shown to be antimutagenic for EMS induction of reversions which occur by a GC to AT transition. Caffeine acts as a comutagen for the induction by EMS of mutant phages and produces a significant increase in the frequency of reversions from rII to r+. Caffeine is slightly mutagenic for the phage strain carrying the wild type polymerase and inhibits the action of the 3' leads to 5' exonuclease function of T4 DNA polymerase as measured in vitro. These findings suggest that caffeine acts by directly influencing nucleotide selection or the editing function of the DNA polymerase.

Caffeine↗

Antimutagens against mutT.

In E. coli strain 91P containing the mutator gene mutT-, caffeine, spermine and quinacrine, but not guanosine, act as antimutagens, reducing the frequencies of mutation from ara- leads to Ara.

Arabinose↗

The effect of streptomycin resistance, caffeine and acriflavine on ultraviolet light-induced reversion to tryptophan independence in strains of Escherichia coli B/r.

When the excision proficient strain E. coli WP2 Hcr+ trp- was grown to stationary phase by glucose starvation in M-9 minimal medium before UV -irradiation, the ability of nutrient broth enrichment of minimal medium to enhance trp- leads to Trp+ reversion was greatly reduced. Less than 50% of the Trp+ revertants were found to be ochre suppressors. However, in the WWP2 Hcr- strain, 75-86% of the tested revertants were ochre suppressors. This indicates that, under the cultural conditions employed, many potential suppressor mutations were removed by excision repair in the presence of broth enrichment. Broth enhancement of reversion also occurred in the Hcr- strain, which indicates that a less error-prone mode of recombination repair functions under minimal growth conditions. An Hcr+ strr derivative of WP2 Hcr+ was more resistant than its strs parent to the lethal effect of UV light and showed a lower UV-induced Trp+ reversion frequency. The percentage of Trp+ revertants that were due to ochre suppressors was markedly reduced in the strr strain. The Hcr- strr strain also had a lower UV-induced Trp+ reversion frequency than its strs parent. The excision repair inhibitor caffeine had little effect at lower UV doses on increasing Trp+ reversion in both Hcr+ strains. Acriflavine, however, was effective at lower UV doses in enhancing reversiin of the Hcr+ strains and the degree of enhancement increased with the dose. Acriflavine appeared to specifically enhance the number of ochre suppressing Trp+ revertants. In both Hcr- strains (strs and strr) caffeine (500 mug/ml) had no effect on survival but reduced the UV-induced Trp+ reversion frequency acting as an antimutagen. In contrast, acriflavine (2 mug/ml) decreased survival and increased the Trp+ reversion frequency of the Hcr- strains. The data on spontaneous Trp+ reversion frequencies show that the Hcr+ strs strain had a higher spontaneous reversion frequency than the Hcr- strs strain on all plating media. Further, caffeine was shown to reduce spontaneous Trp+ reversion in both Hcr+ and Hcr- strains while acriflavine increased the spontaneous reversion frequencies of both strains.

Acridines↗

Isolation and characterization of two cell types of Coxiella burneti phase I.

Two morphologically distinct cell types of Coxiella burneti phase I have been separated on the basis of unique buoyant densities. When centrifuged to equilibrium in cesium chloride or density gradients of sucrose or Renografin, the cells band in two zones. Electron micrographs of ultrathin sections of the two cesium chloride-separated cell types indicate a considerable number of morphological differences. The lower-density cells are small, compact, and rodshaped and have very dense nucleoids. The cell type of highest density is larger, rounded, and more pleomorphic, and the nucleoid filaments are more dispersed. The two cell types are nearly identical in sedimentation rates, and both infect chick yolk sac cells and are lethal to chick embryos. They convert to a mixture of cell types when cultured separately. Treatment with Formalin induces all cells to band at the same position when centrifuged to equilibrium in cesium chloride. The cell type variance was found to be independent of the antigenic phase phenomenon of C. burneti.

Animals↗

Ultrastructure of Nocardia cell growth and development on defined and complex agar media.

The cell growth of Nocardia strain 721-A on Brain Heart Infusion Agar (BHIA), nutrient agar, and chemically defined agar media was studied by light and electron microscopy. Light microscopy revealed a change in cell morphology induced by growth on BHIA. Electron microscopy demonstrated a concurrent change in intracellular complexity. On BHIA, the cells became bulbous and developed irregularly branched filaments which fragmented by multiple and random septation. These fragments appeared to undergo a secondary stage of development similar to that described for Arthrobacter. Cells grown on defined or nutrient agar did not become bulbous and lacked the unusual complexity found in cells grown on BHIA. Intracytoplasmic membranes were altered by the nutritional state of the cell and changed during cell development.

Agar↗

Mechanism of caffeine enhancement of mutations induced by sublethal ultraviolet dosages.

Certain chemical compounds increase mutation frequency of Escherichia coli B/r significantly when used in conjunction with nonlethal ultraviolet (UV) dosages. Studies were done to elucidate the mechanism of this enhancing mutational effect. Dark survival curves showed that 500 mug of caffeine per ml in the postirradiation medium markedly decreased survival to 60 ergs/mm(2) of UV in strain B/r. Caffeine did not markedly decrease survival to UV in strain B/r WP-2 hcr(-). At least 90% of the mutations induced to streptomycin resistance by UV and 85% of those induced by UV with caffeine could be photoreversed. Experiments with thymine analogues suggested that thymine dimerization at the streptomycin locus was the primary premutational photoproduct induced by sublethal UV dosages. Caffeine did not interfere with the photoreversal of induced mutants, indicating that it probably does not bind to the photoreactivating enzyme or to a UV-induced lesion in the DNA. Addition of DNA or irradiated DNA with 500 mug of caffeine per ml resulted in no loss of the caffeine activity. The excision of UV-induced thymine-containing dimers from E. coli B/r T(-) was investigated in the presence and absence of caffeine. Our results indicated that caffeine prevents excision of thymine dimers, presumably by binding to the excising enzyme. This binding results in an impairment of repair, which produces the increase in mutant numbers.

Acridines↗