Salmonella/microsome mutagenicity of monochloro derivatives of some di-, tri- and tetracyclic aromatic hydrocarbons

A series of 8 monochloroarenes have been tested for mutagenicity in the Salmonella/microsome assay. None of the compounds was detectably active in the absence of mammalian activation whereas, depending on structure, some of the compounds were mutagenic in its presence having responses higher than those reported for the parent compounds.     

Mutagenicity of benzylic acetates, sulfates and bromides of polycyclic aromatic hydrocarbons

Studies were performed to determine the direct mutagenicity of the acetates and some bromides and sulfates of hydroxymethyl polycyclic aromatic hydrocarbons in S. typhimurium strains TA98 and TA100. Benzylic acetates, bromides and sulfates were synthesized and characterized. The compounds tested were benzyl alcohol, 5-hydroxymethylchrysene, 1-hydroxymethylpyrene, 6-hydroxymethylbenzo[a]pyrene, 6-(2-hydroxyethyl)benzo[a]pyrene, 6-hydroxymethylanthanthrene, 9-hydroxymethylanthracene, 9-hydroxymethyl-10-methylanthracene, 7-hydroxymethylbenz[a]anthracene, 7-(2-hydroxyethyl)benz[a]anthracene, 12-hydroxymethylbenz[a]anthracene, 7-hydroxymethyl-12-methylbenz[a]anthracene, 12-hydroxymethyl-7-methylbenz[a]anthracene, 1-hydroxy-3-methylcholanthrene, 2-hydroxy-3-methylcholanthrene, 3-hydroxy-3, 4-dihydrocyclopental[cd]pyrene and 4-hydroxy-3, 4-dihydrocyclopental[cd]pyrene. The benzylic sulfate esters of 6-hydroxymethylbenzo[a]pyrene and 7-hydroxymethylbenz[a]anthracene were the most mutagenic compounds, whereas the aliphatic sulfate ester of 7-hydroxyethylbenz[a]anthracene did not cause an increase in mutations above background. All meso-anthracenic benzylic acetate esters were mutagenic in both strains with various degrees of activity, whereas the corresponding non-benzylic esters were inactive, as expected. Of the non-meso-benzylic acetate esters, only the 3-acetoxy-3, 4-dihydrocyclopenta[cd]pyrene was mutagenic. In the benzylic bromide series, only the eight mesoanthracenic were mutagenic, whereas benzyl bromide and 5-bromomethylchrysene were inactive. The aliphatic bromides, 6-(2-bromoethyl)benzo[a]pyrene and 7-(2-bromoethyl)benz[a]anthracene did not display significant activity. The potencies of the acetate esters more accurately reflect the mutagenicity because the rate of solvolysis did not compete with the reactivity of the esters with bacterial DNA. In the case of benzylic sulfates and bromides, the rate of solvolysis was very rapid and could have diminished the level of mutagenicity, depending on the assay conditions. These results demonstrate that meso-anthracenic benzylic acetates, sulfates and bromides are mutagenic, whereas benzylic acetate esters attached to other carbon atoms are inactive.     

Mono- bi- tri- and polypartite models in photosynthesis

Summary It is shown how energy fluxes in mono-bi-tri- and polypartite photosystems can be described. The derivation of the energy distribution term and the probability of spill over p₂₁ as proposed by W.L. BUTLER are reviewed.     

Mutagenicity of marijuana and Transkei tobacco smoke condensates in the salmonella/microsome assay

Extracts and smoke condensates of marijuana, Transkei home-grown tobacco and also commercial cigarette tobaccos were assayed for their mutagenic activity to Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538, both with and without metabolic activation. No mutagenic activity was detected in dichloromethane extracts of marijuana and tobacco per se, but all the smoke condensates exhibited mutagenicity with metabolic activation. The only strain not mutated by any of the pyrolyzates was TA1535. Transkei tobacco pyrolyzate proved to be the most mutagenic, followed by marijuana, pipe and cigarette tobacco. Mutagenicity was positively associated with the nitrogen content of the various products. The potent mutagenic action of marijuana smoke condensate, coupled with a condensate yield of more than 50% higher than that of cigarette and pipe tobacco, indicates a high carcinogenic risk associated with marijuana smoking.     

Lack of mutagenicity of some phytoestrogens in the salmonella/mammalian microsome assay

8 phytoestrogens were tested for mutagenicity using a variation of the Salmonella/mammalian microsome (or Ames) assay. Zearalenone is a mycotoxin produced by a grain contaminant, Fusarium graminearum (Gibberella zeae) and the isomers of zearalanol are reduced derivatives of this compound. The remaining compounds are all flavonoids which occur naturally at relatively high concentrations in many plants, particularly legumes. 4 of these flavonoids (daidzein, genistein, formononetin and biochanin-a) are isoflavones and the 5th, coumestrol, is a coumestan. Each compound was tested at several concentrations ranging from 1--500 micrograms per plate. The microsomal fracton was obtained from Aroclor 1254 (a PCB)-induced rat livers. None of the compounds tested was mutagenic to Salmonella strains TA1538, TA98 or TA100 at any concentration.     

Indirect- and direct-acting mutagenicity of diesel,coal and wood burning-derived particulates and contribution of polycyclic aromatic hydrocarbons and nitropolycyclic aromatic hydrocarbons

Particulates exhausted from two types of diesel engines (DEPs), burning-derived particulates from three types of coal (CBPs) and burning-derived particulates from three types of wood (WBPs) were separated into four fractions by silica-gel column chromatography using n-hexane, n-hexane–dichloromethane (3:1, v/v), dichloromethane and methanol, as the corresponding eluents. The indirect-acting mutagenicity of each fraction was assayed by the Ames test using the Salmonella typhimurium TA100 strain with S9 mix and the direct-acting mutagenicity was assayed using the S. typhimurium TA98 strain without S9 mix. The polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs) of each fraction were determined by high-performance liquid chromatography (HPLC). Both direct- and indirect-acting of mutagenicities were the highest in samples of DEPs. The contributions of PAHs in samples of WBPs and NPAHs in DEPs were the largest, respectively.     

Mutagenicity and polycyclic aromatic hydrocarbons analysis of airborne particulate matters from Taipei City

The mutagenic activity of dichloromethane extracts of 147 air particulate samples collected from 8 stations during December 1986-June 1987 in Taipei city was consistently higher in S. typhimurium strain TA 98 than in strain TA 100 in the presence of S9 mixture. Among the 8 stations, Nan Kang Police Station, Fu Hsing Elementary School, and Chung Hsing University which were located in the industrial district, downtown area, and heavy traffic zone, respectively, had significantly higher levels of PAHs than the other stations. In contrast, the levels of PAHs were much lower in the suburban station, near Pei Tou Elementary School. However, PAH contents of the air particulate samples collected from these stations did not show good correlation with their mutagenicity. The air particulates collected at some stations on Sunday when the traffic changed from heavy to light showed lower mutagenicity and PAH contents as compared with the other weekdays at the same stations. On the contrary, the samples collected at Pei Tou station in a suburban area where the traffic changed from light to heavy on Sunday showed higher mutagenicity and PAH contents. The monthly average of PAHs of air particulate samples collected over a 7-month period from 8 stations in Taipei city was lower than the average in 1980. Moreover, when compared with other countries, such as U.S.A., the Netherlands, West Germany, Italy, Norway, and Japan, the levels of PAHs and mutagenicity of air particulate matters in Taipei city were similar or slightly lower. The mutagenicity and contents of PAHs of air particulates collected from burnt ABS were significantly higher than those of burnt PVC. One sample PT-6-3 was collected while a nearby garbage collection area was on fire. The mutagenicity of that sample increased 3 to 16 fold and contained an 11 to 33 times higher content of the six PAHs (BaP, BeP, BbF, BaA, Chr, and DbA) as compared with the other samples collected at the same location at a different time. The higher mutagenicity and PAH contents of that sample might be due to the pollution of the air from combustion of the garbage containing products made of ABS.     

Mutagenicity of the anticancer drug, caracemide, and related compounds for salmonella

Caracemide, MeCON(CONHMe)(OCONHMe) (I), is a novel anticancer drug. Since it was derived from acetohydroxamic acid (II), a known mutagen, its potential metabolites and related compounds were synthesized and tested for mutagenicities in S. typhimurium TA98 and TA100. These compounds were: MeNHCONH(OCONHMe) (III), MeCONH(OCONHMe) (IV), MeCONOH(CONHMe) (V), MeNHCOONH2 X HCl (VI), MeNHCONHOH (VII), MeNHCOON(CONHMe)2 (VIII), and NOH(CONHMe)2 (IX). The mutagenicities in the absence of rat liver homogenate were: (VI) much greater than (IV) greater than (II), (III), (V). The other compounds were not mutagenic. (I) was mutagenic only in the presence of rat liver homogenate. The doses required to demonstrate mutagenicities of these compounds were from 0.05 to 5 mumoles/plate. The major hydrolytic products at 25 degrees C, pH 7, were (III), (IV), and (V) from (I); (II) and (III) from (IV); and (II), (III), (VII) and MeNHCONH(OCOMe) (X) from (V). (III) was stable at pH 7. Treatment of (IV) with HCl yielded (VI). Hydrolysis of (III) or (V) with ammonia yielded (VII). These results suggest that caracemide may be activated enzymatically or nonenzymatically by deacetylation or decarbamoylation, and its anticancer activity may be related to the reactivity of its metabolites with DNA. The synthetic procedures and characterizations of new compounds (IV), (V) and (X) are described.     

Direct-acting mutagenicity of extracts of coal burning-derived particulates and contribution of nitropolycyclic aromatic hydrocarbons

Benzene-ethanol extracts from particulates produced by coal burning were separated into four fractions by silica-gel column chromatography using n-hexane (240 ml), n-hexane-dichloromethane (3:1, v/v) (200 ml), dichloromethane (200 ml) and methanol (450 ml), as the corresponding eluents. The mutagenicity of each fraction was assayed by the Ames test using the Salmonellatyphimurium YG1024 strain. The nitropolycyclic aromatic hydrocarbons (NPAHs) of each fraction were assayed by high-performance liquid chromatography with chemiluminescence detection. The highest activity was observed in the n-hexane-dichloromethane fraction (Fr. 2). The mutagenic contribution of this fraction was 69.9% of the total of the four fractions. Ten of 11 NPAHs detected were in Fr. 2 and one (1-nitropyrene) was most concentrated in Fr. 3. Among the NPAHs examined, 3-nitrobenzanthrone made the largest mutagenic contribution. This is the first report of detection of 3-nitrobenzanthrone in coal burning-derived particulates.     

Analysis and tracing of polycyclic aromatic hydrocarbons and mutagenicity of airborne particulates from the Taipei area

In a 3-year study, we determined the mutagenicity and polycyclic aromatic hydrocarbons (PAHs) of airborne particulates collected during December 1987-September 1988 (216 samples), October 1988-January 1989 (81 samples), and October 1989-April 1990 (52 samples) from 9 locations in the Taipei area. We found that dichloromethane extracts of all the samples were mutagenic to Salmonella typhimurium in the Ames test. Moreover, the mutagenicity was much higher in the presence of rat liver microsomal fraction (S9 mixture) than that observed in its absence, which indicates that airborne particulates contained both direct and indirect mutagens. The average mutagenicity of the samples collected in the 3-year period was 137, 127, and 118 histidine revertants/10 m3 air, respectively. On the other hand, we found that dichloromethane extracts of each airborne particulate sample contained 14 PAHs with wide variations in concentration and relative distribution. The levels of Pha, Flu, Pyr, and Ben were much higher than the PAHs with higher ring numbers such as BaP, BeP, Pr, IP, and DbA. The average PAH content was 8.0, 5.0, and 7.8 ng/m3 air for airborne particulates collected during December 1986-September 1987, October 1988-January 1989, and October 1989-April 1990, respectively. Among the 9 stations, Fu Hsing Elementary School and Chung Hsing University (Taipei campus), which are, respectively, located in the downtown area and a heavy traffic zone, had significantly higher levels of mutagenicity and PAHs than did the other stations. Moreover, comparative analysis of PAH levels of airborne particulates over the 3-year period revealed an interesting season-dependent change of PAH content in airborne particulates from the Taipei area. The concentrations of individual and total PAHs were consistently lower in the summer than those in the winter. A similar pattern of seasonal change was also observed in the mutagenicity of airborne particulate samples examined. It is worth mentioning that neither PAH level nor mutagenicity of airborne particulates showed significant yearly change over the 3-year period of study. As part of an effort to identify pollution sources, we examined the mutagenicity and PAH compounds of air particulates collected from the burning of garbage (14 samples) and motor-vehicle exhaust in the Hsin Hai Tunnel (17 samples), Taipei. The results showed that garbage burning gave rise to air particulates containing several hundred times higher levels of PAHs and about 20 times stronger mutagenicity, while the motor-vehicle exhaust contained about ten times higher PAH content and mutagenicity as compared with those of airborne particulates of the Taipei city.(ABSTRACT TRUNCATED AT 400 WORDS)     

Enzymatic characterization of the polynuclear aromatic hydrocarbons activating rat-liver preparations used in the mutagenicity test of Ames

Stability studies were performed on the mono-oxygenase system involved, in particular, in the activation of polynuclear aromatic hydrocarbons (PAHs) present in rat-liver preparations used in the Ames mutagenicity test. The results indicated a good stability of the spectral response of the cytochrome-P-450 system, but a much lower stability of its enzymatic activities measured with various substrates, thus showing the inadequacy of the spectral response to characterize the PAH mono-oxygenase activity of the preparations. Epoxide hydrolase activity was found to be stable. Various mono-oxygenase activities were measured in preparations induced with phenobarbital, 3-methylcholanthrene or Aroclor 1254. The activities of two enzymes, benzo[a]pyrene hydroxylase and ethoxyresorufin-O-dealkylase, were found suitable to characterize the capacity of the preparations to metabolize PAH to mutagens. The efficiency of the same preparations to promote the mutagenicity of benzo[a]pyrene and aflatoxin B1 in the Ames test was determined. There was an excellent general correlation between the efficiencies for mutagenic activation of the preparations and the two enzymatic activities mentioned above. Determination of ethoxyresorufin-O-dealkylase (or benzo[a]pyrene hydroxylase) and benzo[a]pyrene 4,5-oxide hydrolase activities is proposed for characterizing the rat-liver preparations used in the Ames test.     

Mutagenicity of aryl propylene and butylene oxides with salmonella

10 aryl propylene oxides and 6 aryl butylene oxides were synthesized. Dose-mutagenicity relationships were studied for these compounds and for 1,2-epoxybutane, using both the preincubation and plate incorporation Ames tests with Salmonella typhimurium strains TA100 and TA1535. Structure-mutagenicity relationships were further examined by concurrent testing at single doses with the plate incorporation assay in strain TA100. In both series of compounds, mutagenicity showed very correlation to chemical reactivity, molar volume and partition values. However, all compounds were mutagenic in at least one system with the propylene oxides being more mutagenic than the corresponding butylene oxide derivatives. The naphthyl derivatives in each series were the most mutagenic.     

Radiation-induced mutagenicity and lethality in ames tester strains of salmonella

Mutation and killing induced by X radiation and 60CO gamma radiation were studied in six different histidine-requiring auxotrophs of Salmonella typhimurium. Strain TA100, which is sensitive to base-pair substitutions, and strains TA2637 and TA98, which are sensitive to frameshifts, carry the pKM101 plasmid and exhibit significantly higher radiation-induced mutations compared to their plasmidless parent strains TA1535, TA1537, and TA1538, respectively. Among the plasmid-containing strains, TA98 and TA2637 are much more sensitive to the mutagenic action of radiation than is TA100 based on a comparison with their respective spontaneous mutation rates; however, no uniformity was observed in the responses of the strains to the lethal action of ionizing radiation. The pKM101 plasmid provides partial protection against lethality in TA100 and TA2637, whereas the same plasmid enhances the lethal action of ionizing radiation in TA98. The following conclusions are consistent with these observations: (1) the standard Ames Salmonella assay correctly identifies ionizing radiation as a mutagenic agent; (2) frameshift-sensitive parent strains are more sensitive to the mutagenic effects of ionizing radiation than is the only strain studied that is sensitive to base-pair substitutions; and (3) enhancement of mutagenesis and survival is related to plasmid-mediated repair of DNA damage induced by ionizing radiation and does not involve damage induced by Cerenkov-generated uv radiation which is negligible for our irradiation conditions.