Environ Mutagen, 1984, 6(4), 497 - 515 Microbial mutagenicity of isomeric two-, three-, and four-ring amino polycyclic aromatic hydrocarbons; Later DW et al.; The isomers of various two-, three-, and four-ring amino polycyclic aromatic hydrocarbons were tested for mutagenic activity using a microbial plate incorporation test with four Salmonella typhimurium strains (TA98, TA100, TA1535, and TA1537) . All compounds were assayed with an S9 metabolic activating enzyme system . The two-ring compounds were tested only with TA98 . All were weakly mutagenic (1-10 rev/micrograms) except 2-aminobiphenyl, which was not mutagenic under these test conditions . All except two of the 13 fused three-ring compounds (aminofluorenes, aminoanthracenes, and aminophenanthrenes) were active frame shift mutagens; only the aminophenanthrenes were active base-pair mutagens . The potency of this group of isomeric compounds ranged from moderately (approximately 20 rev/microgram) to strongly (greater than 5,000 rev/microgram) mutagenic . As a group, the pericondensed four-ring amino compounds were the most mutagenic of the three groups tested . All of the aminofluoranthene and aminopyrene isomers showed significant mutagenic activity with TA98, TA100, and TA1537 . In general, the mutagenic potency of the amino polycyclic aromatic compounds tested was highly dependent on the structural position of the amino group. J Cancer Res Clin Oncol, 1984, 108(1), 76 - 80 Alcoholdehydrogenase as an activating enzyme for N-nitrosodiethanolamine (NDELA): in vitro activation of NDELA to a potent mutagen in Salmonella typhimurium; Eisenbrand G et al.; N-Nitrosodiethanolamine (NDELA), a potent carcinogen, has not so far been found to be mutagenic in a wide range of test systems . In particular, mutagenicity testing in Salmonella typhimurium with rat liver S-9 mix or microsomal fraction used for activation has failed to indicate mutagenicity . However, when incubated with alcohol dehydrogenase (ADH) in the presence of NAD, NDELA is converted to a potent mutagen . A possible mechanism of activation comprises the generation of an aldehyde as a primary metabolite formed by NAD/ADH and its subsequent rearrangement into cyclic intermediates . The latter might either be further metabolized or spontaneously decompose into various alkylating agents and glycolaldehyde . Standard test conditions used for the Ames test will not favor the detection of mutagens to be activated by NAD/ADH because they require the presence of NADPH, whereas ADH needs NAD to become an activating enzyme, as shown for NDELA. Environ Mutagen, 1984, 6(3), 343 - 54 Mutagenicity screening of foods . II . Results with fruits and vegetables; Stoltz DR et al.; A survey of the mutagenic potential of a wide variety of food products has been initiated with results for 28 different beverages reported previously {Stoltz et al, 1982b} . Here, results for samples of 46 widely consumed fruits and vegetables from six general categories are given . Each sample was concentrated and fractionated by polarity and solubility to give five fractions, each of which was assayed for mutagenic potential with Salmonella typhimurium TA98 and TA100 . Although statistical analysis of the data resulted in positive findings for 22 fruit and vegetable samples, only six products (grapes, onions, peaches, raisins, raspberries, strawberries) demonstrated potent mutagenic activity. Environ Mutagen, 1984, 6(3), 311 - 20 Chromium (VI) potentiates mutagenesis by sodium azide but not ethyl methanesulfonate; LaVelle JM et al.; A fluctuation test using Salmonella typhimurium strain 1535 has been used in an experimental protocol to assess biological effects of interactions between chromium (VI), such as K2CrO4, and two DNA-damaging agents, ethyl methanesulfonate (EMS), and sodium azide . Mutagenicity, expressed as the average number of mutations induced over a parallel control, was determined for the compounds alone and in combination . The significance of the differences between the "expected" response, calculated by simple addition of the responses from the individual tests, and the observed response when the combination was tested, were estimated by chi square . For the combination of K2CrO4 and NaN3, the response was significantly greater than expected suggesting a possible potentiation of mutagenesis . The opposite (a less-than-additive response) was found for the K2CrO4/EMS combination . Both effects were found to be dose related to the concentration of potassium chromate used . Toxicity of the compounds or their combinations to the bacteria could not explain the results. Drug Chem Toxicol, 1984, 7(3), 243 - 57 Evaluation of the mutagenic potential of corn (Zea mays L.) grown in untreated and atrazine (AAtrex) treated soil in the field; Sumner DD et al.; Bacterial assays using extracts from field corn plants (harvested at one month, silage and mature stages) do not indicate that soil treatment with atrazine, at its maximum use rate, alters the endogenous mutagens present in these extracts, nor that atrazine itself is degraded to mutagenic products . Extracts of corn grown in soil treated with AAtrex were equally mutagenic with those of corn grown in untreated soil when tested in Salmonella typhimurium TA-100 by a reversion assay or in Salmonella typhimurium TM-677 in a forward mutation assay . Higher concentrations of histidine in corn grown in AAtrex treated soil may interfere with the reversion assay, but do not affect the forward mutation assay . The nature of the agent(s) responsible for the positive response was not determined . The mutagenicity may be due to natural plant constituents, an artifact of the sample preparation, or mycotoxins from some unrecognized plant infection . The experimental results in these field studies do not show that atrazine is degraded or metabolized by corn plants to mutagens in this sensitive bacterial assay. Mol Gen Genet, 1984, 194(1-2), 219 - 26 Methionine and glutamine transport systems in D-methionine utilising revertants of Salmonella typhimurium; Poland J et al.; In Salmonella typhimurium, methionine auxotrophs such as metB can use D-methionine as a methionine source . MetP mutations prevent this growth since D-methionine can enter only via the metP high-affinity methionine transport system . D-methionine utilising revertants ( Dmu +) were selected from metB23 metP760 ( HU76 ) following nitrosoguanidine mutagenesis . The properties of two such revertants, HU206 and HU415 , indicated that reversion was not due to backmutation of the metP760 mutation . Genetic analysis indicated that each strain possessed two mutations, designated dmu and gln, in addition to the original metB23 and metP760 mutations . The dmu mutation restores ability to grow on D-methionine, partly restores D- and L-methionine transport activity, and makes the cells particularly sensitive to inhibition by L-glutamine while growing on D but not L-methionine . The growth inhibition by L-glutamine was shown to be caused by competition by L-glutamine for D-methionine transport by the high-affinity methionine system . The gln mutation greatly reduces activity of the high-affinity glutamine transport system . The Dmu + strains are also partly defective in the glutamine low-affinity transport system, possibly because the partially-restored methionine high-affinity system, or a component of tis system, functions in the transport of glutamine by its low-affinity system. Gann, 1984 Jan, 75(1), 8 - 16 Comparison of mutagenicities of N-nitrosamines on Salmonella typhimurium TA100 and Escherichia coli WP2 uvrA/pKM101 using rat and hamster liver s9; Araki A et al.; The mutagenicities of twelve N-nitrosamines were tested on Salmonella typhimurium TA100 and Escherichia coli WP2 uvrA/pKM101 in the presence of rat liver S9 or hamster liver S9 by "preincubation" and "pour-plate" assays . The following eleven N-nitrosamines were mutagenic: N-nitroso-N-dimethylamine; N-nitroso-N-diethylamine; N-nitroso-N-di-n-propylamine; N-nitroso-N-di-n-butylamine; N-nitroso-N-methyl-n- amylamine ; N-nitroso-piperidine; N-nitrosomorpholine; N-nitroso-N-methyl-piperazine; N-nitroso-N-methyl-benzylamine; N-nitroso-N-methyl- phenylamine and N-nitroso-N-phenyl-benzylamine . N-Nitroso-N-diphenylamine was not mutagenic . E . coli WP2 uvrA/pKM101 was more sensitive than S . typhimurium TA100 to the mutagenic actions of N-nitroso-dialkyl amines, N-nitroso- aklyl -aryl amines and N-nitroso- diaryl amines, but S . typhimurium TA100 was more sensitive to those of N-nitroso-cyclic amines . Hamster liver S9 was better than rat liver S9 for metabolic activation of N-nitrosamines, and the preincubation step enhanced the mutagenicities of N-nitrosamines. Gann, 1984 Jan, 75(1), 1 - 3 Tyramine is a major mutagen precursor in soy sauce, being convertible to a mutagen by nitrite; Ochiai M et al.; Tyramine was identified as a new mutagen precursor in Japanese soy sauce, becoming mutagenic after treatment with nitrite under acidic conditions . The mutagenic compound was identified as 4-(2-aminoethyl)-6-diazo-2,4- cyclohexadienone , and its specific mutagenic activity was 112 revertants/micrograms towards Salmonella typhimurium TA100 without S9 mix. Int Arch Occup Environ Health, 1984, 53(4), 359 - 64 Lack of mutagenic activity of white spirit; Gochet B et al.; The mutagenic potential of white spirit, a typical mixture of primarily aliphatic hydrocarbons used as a solvent, was investigated using a battery of test systems . The ability of this compound to induce gene mutations was assayed by the Ames' test with different strains of Salmonella typhimurium; its potential clastogenicity was tested in vivo on mouse bone marrow cells; in vitro induction of sister chromatid exchanges was studied in human lymphocytes . Negative results were obtained in all test systems . It is concluded that, in spite of its evident toxicity, white spirit does not display mutagenic properties. Arch Microbiol, 1984 Jan, 137(1), 70 - 3 Transport and metabolism of trehalose in Escherichia coli and Salmonella typhimurium; Marechal LR; The metabolism of trehalose in wild type cells of Escherichia coli and Salmonella typhimurium has been investigated . Intact cells of Escherichia coli (grown on trehalose) accumulated {14C}-trehalose as {14C}-trehalose 6-phosphate . Toluene-treated cells catalyzed the synthesis of the {14C}-sugar phosphate from {14C}-trehalose and phosphoenolpyruvate; ATP did not serve as phosphoryl donor . Trehalose 6-phosphate could subsequently be hydrolyzed by trehalose 6-phosphate hydrolase, an enzyme which catalyzes the hydrolysis of the disaccharide phosphate into glucose and glucose 6-phosphate . Both Escherichia coli and Salmonella typhimurium induced this enzyme when they grew on trehalose . These findings suggest that trehalose is transported in these bacteria by an inducible phosphoenolpyruvate:trehalose phosphotransferase system . The presence of a constitutive trehalase was also detected. Environ Mutagen, 1984, 6(2), 145 - 51 Mutagenicity of some benzidine congeners and their N-acetylated and N,N'-diacetylated derivatives in different strains of Salmonella typhimurium; Reid TM et al.; The Ames Salmonella/microsome test was used to compare the mutagenic response of Salmonella typhimurium TA100, TA98, TA1538, and TA1535 to 12 benzidine derivatives, ie, benzidine, 3,3'-dimethoxybenzidine, 3,3'-dimethylbenzidine, 3,3'-dichlorobenzidine, and the corresponding N- and N,N'-diacetylated derivatives . With a few exceptions, the mutagenic response to this series of compounds varied in the order TA98 greater than TA1538 greater than TA100 greater than TA1535 = 0, and the N-monoacetylated derivatives were more mutagenic than either the parent diamines or the N,N'-diacetyl derivatives . The relative mutagenicities of the parent amines for TA98 were 3,3'-dichlorobenzidine much greater than 3,3'-dimethoxybenzidine greater than benzidine greater than 3,3'-dimethylbenzidine. Environ Mutagen, 1984, 6(2), 131 - 44 Contribution of nitropyrene to the mutagenic activity of coal fly ash; Harris WR et al.; Stack-collected coal fly ash from western low-sulfur coal was extracted with 60:40 benzene/methanol . This extract was fractionated by preparative-scale high performance liquid chromatography (HPLC) and the mutagenic activity of 14 fractions was evaluated by microbial assay with Salmonella typhimurium TA1538 . A widespread distribution of direct-acting mutagens, which probably includes both mono- and dinitroaromatics, was detected . HPLC methods were also used to isolate 1-nitropyrene from the total benzene/methanol extract . The identification of 1-nitropyrene was based on gas chromatographic and HPLC retention measurements and mass spectral data . The concentration of 1-nitropyrene in the ash extract was determined by quantitative HPLC analyses . Mutagenicity assays of the total extract and an authentic 1-nitropyrene standard with Salmonella strains TA1538, TA100, and TA98 indicated that the 1-nitropyrene accounts for approximately 0.03-0.16% of the total mutagenic activity of the extract. Am J Vet Res, 1984 Jan, 45(1), 59 - 66 Aromatic-dependent Salmonella typhimurium as modified live vaccines for calves; Smith BP et al.; Strains of Salmonella sp with complete nonreverting aromatic biosynthesis (aro) defects are expected to be nonvirulent, in respect to invasive infection, because they need the aromatic metabolites paraaminobenzoate (for making folate) and dihydroxybenzoate (for making enterochelin) which are not available in host tissues . Derivatives with transposon-generated complete nonreverting aro-defects were prepared from 3 mouse-virulent strains of S typhimurium, namely, FIRN, WRAY, and UCD . The latter 2 parent strains originally were isolated from calves and are known to be calf-virulent . The resultant aromatic-dependent (aro-) strains were used to vaccinate 27 calves (2 to 3 weeks old), usually giving 2 doses by the IM route (10(9) bacteria) or orally (1.5 X 10(11)) . Vaccination did not cause severe ill effects in any calf . Thus aro- defects cause loss of virulence for calves, as previously shown for mice . Vaccinated and control calves were challenge exposed, usually at 5 weeks of age, by feeding 1.5 X 10(11) cells of 1 of 2 calf-virulent S typhimurium strains, either UCD 108-11 or SL1323 . Of the 16 challenge-exposed control calves, all became anorectic and depressed (CNS), and 15 had diarrhea . Fourteen of the 16 died; all tested tissues were bacteriologically culture-positive for Salmonella at necropsy . Vaccination with the live UCD aro- vaccine strain, SL1479 by either of 2 schedules (IM or orally) appeared effective.(ABSTRACT TRUNCATED AT 250 WORDS) Toxicology, 1984 Jan, 29(3), 261 - 70 Quantitative correlation between the metabolism and the mutagenic activity of N-nitrosopyrrolidine; Gilbert PJ et al.; The metabolism of N-nitrosopyrrolidine (NPyrr) via alpha-hydroxylation is modified by pretreatments of the animals with compounds which affect the microsomal level of cytochrome P-450 and by addition, in vitro, of 2-diethylaminoethyl-2,2-diphenyl valerate hydrochloride (SKF 525-A), an inhibitor of cytochrome P-450 . This phenomenon is due exclusively to the induction or the inhibition of the enzymatic activity involved in the microsomal metabolism . After preincubation in liquid medium, the mutagenic activity of NPyrr towards the Salmonella typhimurium strain TA 1530 is similarly modified by these effectors . A similar effect is not observed when using the plate incorporation method . The mutagenic intermediate is formed by the microsomal fraction . The presence of the S . typhimurium strain TA 1530 decrease the transformation of NPyrr into its ultimate metabolite (1,4-butanediol); there is a relationship between the formation of 1,4-butanediol and the mutagenic activity of NPyrr . The S . typhimurium strain TA 1530 is able to partially transform 4-hydroxybutanal, the first identifiable microsomal metabolite of NPyrr, into its ultimate metabolite (1,4-butanediol). Mutat Res, 1984 Jan, 135(1), 31 - 47 The hair-dye reagent 2-(2',4'-diaminophenoxy)ethanol is mutagenic to Salmonella typhimurium; Venitt S et al.; A new hair-dye ingredient, 2-(2',4'-diaminophenoxy)ethanol (2,4-DAPE), was described as being devoid of any genotoxic activity on the basis of a multi-laboratory study . Since 2,4-DAPE is a close analogue of 2,4-diaminoanisole (2,4-DAA), which is mutagenic and carcinogenic, we tested this claim by assaying 2,4-DAPE for bacterial mutagenicity . Two samples of 2,4-DAPE X 2HCl were synthesized by reduction of the corresponding dinitrophenoxyethanol and identity and purity were established by elemental analysis, NMR spectrometry, mass-spectrometry, UV-spectrophotometry, TLC and HPLC . Fresh aqueous solutions of 2,4-DAPE X 2HCl were assayed in several separate plate tests using S . typhimurium TA1538, TA97, TA98 and TA100, and E . coli WP2uvrA (pKM101), 3 plates per dose and 0%, 4%, 10% and 30% Aroclor 1254-induced rat-liver S9 in S9 mixes . We obtained negative results in TA100 and E . coli . Reproducible, statistically significant dose-related increases in revertants (up to 14 times the background) were obtained in frame-shift mutants of S . typhimurium in the dose range 10-80 micrograms per plate . Mutagenicity was S9-dependent, significant increases in revertants being obtained only with 50 microliter per plate or more of S9 . 2,4-DAPE induced significant mutagenic effects at doses of less than 1 micrograms per ml in TA1538 and TA98 in fluctuation tests using 2% S9 in the S9 mix . In plate tests, 2,4-DAPE was less mutagenic (by a factor of about 8) than 2,4-DAA, which gave the highest mutant yields with 20 microliter S9 per plate (4% S9 in the S9 mix) . 2,4-DAPE obtained commercially was about 8 times more mutagenic than our sample of 2,4-DAPE . After purification, the commercial product, now chromatographically identical with our own sample, gave plate-test results close to those obtained for our samples of 2,4-DAPE . A review of the published reports (in which 2,4-DAPE was claimed to be inactive in a variety of short-term tests) revealed: (a) the use of protocols for bacterial mutagenicity testing which, in the light of our own results, were probably too limited in scope, especially in the choice of conditions for metabolic activation; (b) insufficient information on the identification and purity of the samples of 2,4-DAPE tested in the published collaborative study. Mutat Res, 1984 Jan, 135(1), 21 - 9 Some chloro derivatives of polynuclear aromatic hydrocarbons are potent mutagens in Salmonella typhimurium; Colmsjo A et al.; A series of chlorinations of some polynuclear aromatic hydrocarbons (PAH) were carried out and the products were tested for mutagenicity on Salmonella typhimurium TA98 and TA100 . We conclude that the chlorination of certain PAHs with low mutagenicity, such as pyrene and benzo{e}pyrene, resulted in the formation of two types of product . The chlorination of pyrene was studied in some detail . The major products of this chlorination were chloro-substituted pyrenes . These compounds showed an S9-dependent mutagenicity and were identified as 1-chloro-, 1,6-dichloro-, 1,8-dichloro- and 1,3-dichloropyrene . On tester strain TA100 the mutagenic effect ranged from 1.4 to 14 revertants/nmol, 1,3-dichloropyrene being the most potent of the isomers . Minor products eluting from a chromatograph in a more polar fraction than the major products were also formed . These compounds were less stable than the major products and were identified as pyrene with chloro additions in the 4- and 5-positions, with various chloro substituents at other positions . These minor products showed a high mutagenic effect on Salmonella in the absence of S9 . The mutagenic effect on strain TA100 ranged from 10 to 15 revertants per ng which is at least 40 and 4000 times higher than for 1-nitropyrene and pyrenequinones, respectively . These unstable chloro derivatives of pyrene are difficult to analyse chemically because they are easily degraded and give rise to the more stable 4-chloropyrene. Mol Gen Genet, 1984, 193(2), 332 - 9 Bifunctionality and polarized infidelity at the hisB locus of Aspergillus nidulans; Millington Ward AM et al.; The histidine (hisB) locus of Aspergillus nidulans is unusual in two ways . Firstly, it is bifunctional; besides coding for imidazole glycerol phosphate (IGP) dehydrase, it is required for the production of ascospores (fertility) . It appears, therefore, to be partly homologous to the hisB locus of Salmonella typhimurium, which codes for IGP dehydrase and histidinol phosphate phosphatase . Secondly, during meiosis it is often inaccurately transmitted to the progeny (infidelity) . This phenomenon may be akin to the aberrant recombination events which cause Bar reversion in Drosophila, "selfing" in Salmonella and Neurospora, and gene fusions of the haemoglobin lepore type . A molecular model is proposed to account for the results. Arch Oral Biol, 1984, 29(1), 59 - 63 The bone-resorbing activities in tissue culture of lipopolysaccharides from the bacteria Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Capnocytophaga ochracea isolated from human mouths; Iino Y et al.; The activities of lipopolysaccharides (LPS) were assessed by measuring the calcium release from mouse calvaria in vitro and compared to that of LPS from Salmonella typhimurium . Stimulation of bone resorption was maximal at an LPS concentration of 10 micrograms/ml and at this dose all oral LPS preparations showed similar levels of activity and less than that of LPS from S . typhimurium . Only S . typhimurium LPS and B . gingivalis LPS retained bone-resorbing activity at 0.1 microgram/ml . No bone-resorbing activity was observed against killed bone and histochemical observations of stable acid phosphatase activity indicated both mononuclear and multinuclear cells participating in bone removal . Addition of indomethacin to the culture medium did not inhibit calcium release from the bones by any of the LPS preparations except for that from A . actinomycetemcomitans . Fetal calf serum completely blocked the activities of all the LPS preparations whereas human serum did not inhibit the action of B . gingivalis LPS . Thus this particular LPS could be important in mediating bone loss in chronic periodontitis. Mutat Res, 1984 Jan, 125(1), 95 - 104 Comparison of the frequency of diphtheria toxin and thioguanine resistance induced by a series of carcinogens to analyze their mutational specificities in diploid human fibroblasts; Aust AE et al.; The mutagenic specificities of ethylnitrosourea (ENU), X-rays (+/-)7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7, 8,9,10-tetrahydrobenzo{a}pyrene (BPDE), ICR-191, and N-acetoxy-2-acetylaminofluorene (N-AcO-AAF) were analyzed and compared in diploid human fibroblasts and Salmonella typhimurium . In the human fibroblasts, we compared the frequency of diphtheria toxin (DT)-resistant mutants, presumably induced in the gene coding for elongation factor-2, with the frequency of 6-thioguanine (TG) resistance induced by mutations in the gene coding for hypoxanthine(guanine)phosphoribosyltransferase (HPRT) . Recovery of DT-resistant (DTr) cells requires that the mutant EF-2 retain the ability to carry on protein synthesis since the normal EF-2 will be inactivated by DT selection . Therefore, the DTr mutation cannot involve major changes in the gene . In contrast, cells can acquire TG resistance by any mechanism which eliminates HPRT activity, e.g., base substitution, frameshift, deletion, loss of chromosomes . Each agent was assessed by calculating the ratio of the slopes of the dose-response plots (induced variant frequency as a function of dose of the agent used) for the two markers (DTr/TGr variants.) . In S . typhimurium we examined the reversion frequency in four histidine-requiring strains bearing forward mutations of the frameshift (TA1538, TA98) or missense (TA1535, TA100) type . ENU, which was predominantly a base substitution mutagen in the bacteria, gave a ratio of DTr to TGr variants of 1.5 . As expected of an agent inducing gross chromosomal changes, X-rays induced no revertants in bacteria and in human cells gave a ratio of 0.1 . ICR-191 which was predominantly a frameshift mutagen in bacteria gave a ratio of 0.15 . In the set of bacterial strains containing the plasmid pKM101, BPDE reverted both frameshift and base substitution mutations . It did not cause reversions in the other set of strains . In human cells BPDE gave a response similar to ENU, i.e., a ratio of DTr/TGr variants of 1.5 . As reported by others, N-AcO-AAF was predominantly a frameshift mutagen in bacteria . However, in the human cells it gave a ratio of DTr/TGr variants of 1.5, similar to ENU and BPDE . These results suggest that in human cells, BPDE and N-AcO-AAF, like ENU, yield predominantly base substitutions, while ICR-191 and X-rays largely produce mutations by mechanisms which result in more extensive alterations in the gene. Mutat Res, 1984 Jan, 125(1), 23 - 31 Liver, kidney and small-intestine microsomal-mediated mutagenicity of carcinogenic aromatic amines; Fouarge M et al.; The mutagenicity of 2-aminofluorene, 4-aminobiphenyl and 3,2'-dimethylaminobiphenyl towards Salmonella typhimurium was studied in the presence of microsomes from liver, kidney and small intestine of untreated and pretreated rats . The aim was to study a possible correlation between the organotropism of these amines and their activation into mutagenic intermediates by these three tissues . Pretreatment of the rats with phenobarbital, Aroclor 1254 and 3-methylcholanthrene injected intraperitoneally increased the liver microsomal-mediated mutagenic activity of the three amines but remained without effect on the activating capacity of microsomes from the kidney and small intestine . However, pretreatment with 3-methylcholanthrene administered intragastrically increased the small-intestine microsomal-mediated mutagenicity of 2-aminofluorene almost 3-fold but remained without effect on the mutagenicity of 4-aminobiphenyl and 3,2'-dimethylaminobiphenyl . No mutagenic effect was observed with 4-aminobiphenyl in the presence of kidney microsomes or with 4-aminobiphenyl and 3,2'-dimethylaminobiphenyl in the presence of small-intestine microsomes, obtained from either untreated or pretreated animals . It is concluded that no relationship exists between the mutagenic activities of the three amines, as detected in the Ames test, and their carcinogenic organotropisms. J Immunol, 1984 Jan, 132(1), 347 - 53 In vitro stimulation of C3H/HeJ spleen cells and macrophages by a lipid A precursor molecule derived from Salmonella typhimurium; Vogel SN et al.; C3H/HeJ mice possess a genetic lesion that renders them significantly less responsive to the biologic effects of protein-free lipopolysaccharide (LPS) preparations, and more specifically, to the lipid A region of the LPS molecule . The in vivo manifestations of this mutation are also reflected in vitro in that cells derived from this mouse strain fail to respond to LPS when compared with cells derived from fully endotoxin-responsive mouse strains . The precise nature of this gene defect has not yet been established . In this study, we have examined in vitro the biologic activities of a structurally less complex "lipid A precursor" molecule, produced by a conditionally lethal, temperature-sensitive mutant of Salmonella typhimurium . In contrast to the intact LPS or wild-type lipid A extracted from the parental strain of Salmonella typhimurium, the lipid A precursor induced a highly significant, polymyxin B-inhibitable mitogenic response in splenic cultures derived from LPS-hyporesponsive C3H/HeJ and C57BL/10ScN (nu/nu) mice . In addition, the lipid A precursor was found to stimulate cultures of C3H/HeJ macrophages to produce significant levels of both interleukin 1 (IL 1, previously referred to as "lymphocyte activating factor" or "LAF") and prostaglandins of the E series (PGE) . These findings suggest the possibility that the defect in endotoxin responsiveness exhibited by C3H/HeJ mice may be related to a defect in the processing of wild-type lipid A or LPS to a suitably stimulatory form that is structurally related to the lipid A precursor molecule. J Bacteriol, 1984 Jan, 157(1), 171 - 8 Cloning and characterization of gdhA, the structural gene for glutamate dehydrogenase of Salmonella typhimurium; Miller ES et al.; Glutamic acid is synthesized in enteric bacteria by either glutamate dehydrogenase or by the coupled activities of glutamate synthase and glutamine synthetase . A hybrid plasmid containing a fragment of the Salmonella typhimurium chromosome cloned into pBR328 restores growth of glutamate auxotrophs of S . typhimurium and Escherichia coli strains which have mutations in the genes for glutamate dehydrogenase and glutamate synthase . A 2.2-kilobase pair region was shown by complementation analysis, enzyme activity measurements, and the maxicell protein synthesizing system to carry the entire glutamate dehydrogenase structural gene, gdhA . Glutamate dehydrogenase encoded by gdhA carried on recombinant plasmids was elevated 5- to over 100-fold in S . typhimurium or E . coli cells and was regulated in both organisms . The gdhA promoter was located by recombination studies and by the in vitro fusion to, and activation of, a promoter-deficient galK gene . Additionally, S . typhimurium gdhA DNA was shown to hybridize to single restriction fragments of chromosomes from other enteric bacteria and from Saccharomyces cerevisiae. J Bacteriol, 1984 Jan, 157(1), 100 - 8 Hook-associated proteins essential for flagellar filament formation in Salmonella typhimurium; Homma M et al.; The hooks of the flagella of Salmonella typhimurium were purified by a newly developed method, using a flaL mutant without a filament, and the hook components were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . As a result, we detected three protein species in addition to hook protein . We call these three proteins hook-associated proteins (HAPs) . Their molecular weights were 59,000 for HAP1, 53,000 for HAP2, and 31,000 for HAP3 . The HAP1/hook protein/HAP3/HAP2 molar ratio, calculated from their relative amounts and their molecular weights, was 1:10:1.1:0.53 . The compositions of HAPs were analyzed in the hooks from the other filamentless mutants which were defective in H1 H2, flaV, flaU, or flaW . Hooks from the H1 H2 mutant had the same HAP composition as hooks from the flaL mutant . Hooks from the flaV mutants contained HAP1 and HAP3 . Hooks from the flaU mutants contained HAP1 . Hooks from the flaW mutants contained a very small amount of HAP3 . From these results, the process of hook morphogenesis and the genes responsible for each step were postulated . Electron micrographs of hooks from the filamentless mutants showed that hooks which contained all three HAPs had a sharp clawlike tip, whereas hooks lacking any HAP had a flat tip . Electron micrographs of hooks treated with antibody against the hook protein showed that each claw-shaped end was not covered with antibody . These results strongly suggest that all three HAPs or at least some of them are located at the claw-shaped end and play an essential role in filament formation. Gene, 1984 Jan, 27(1), 47 - 54 Cloning and characterization of the gene for Salmonella typhimurium serine hydroxymethyltransferase; Urbanowski ML et al.; A plasmid containing the glyA gene of Salmonella typhimurium LT2 was constructed in vitro using plasmid pACYC184 as the cloning vector and a lambda gt7-glyA transducing phage as the source of glyA DNA . The recombinant plasmid (pGS30) contains a 10-kb EcoRI insert fragment . Genetic and biochemical experiments established that the fragment contains a functional glyA gene . From plasmid pGS30 we subcloned a 4.4-kb SalI-EcoRI fragment containing the glyA gene and its neighboring regions (plasmid pGS38) . The location and orientation of the glyA gene within the 4.4-kb insert fragment was determined in four ways: (1) comparison of the physical map of the 4.4-kb SalI-EcoRI fragment with the physical map of a 2.6-kb SalI-PvuII fragment that carries the Escherichia coli glyA gene; (2) deletion analysis; (3) transposon Tn5 insertional inactivation experiments; (4) deoxyribonucleic acid sequencing and comparison of the S . typhimurium DNA sequence with the E . coli DNA sequence . A presumptive glyA-encoded polypeptide of Mr 47000 was detected using plasmid pGS38 as template in a minicell system, but not when the glyA gene was inactivated by insertion of a Tn5 element. Scand J Infect Dis, 1984, 16(1), 111 - 9 Effects of Streptococcus pneumoniae, Salmonella typhimurium and Francisella tularensis infections on oxidative, glycolytic and lysosomal enzyme activity in red and white skeletal muscle in the rat; Friman G et al.; Since opinions differ as to whether the oxidative and glycolytic capabilities of skeletal muscle are altered in acute infection, enzyme activities in oxidative, glycolytic and degradative (acid hydrolases) pathways and total protein and DNA were determined in skeletal muscle of rats infected with Streptococcus pneumoniae, Salmonella typhimurium or Francisella tularensis . Studies were performed separately in red (slow twitch) and white (fast twitch) muscle tissue because these fibers function during different types of exercise . In the salmonella- and tularemia-infected rats, the intramitochondrially located oxidative enzymes of muscle were decreased to 56-83% of controls whereas the glycolytic enzyme situated in the cytosol showed an earlier and more pronounced loss of activity, 30-75% of controls . In the pneumococcal infection, only reduced glycolytic activity was significant . DNA concentrations were unchanged in any infection . Reductions during tularemia were statistically correlated with whole-cell protein degradation, while that of the glycolytic enzyme was parallelled by activation of lysosomal enzymes . Red and white muscle tissues responded similarly, in contrast to several other pathologic states that involve a catabolic component of muscle with a predominant response (or damage) in one or the other fiber type. J Bacteriol, 1984 Jan, 157(1), 158 - 64 Isolation and characterization of the Salmonella typhimurium LT2 xylose regulon; Ghangas GS et al.; Salmonella DNA was partially digested with EcoRI, and the digest was fractionated to obtain fragments larger than 8 kilobases (kb) . These were ligated into EcoRI-cut pBR322, and the mixture was used to transform Salmonella Xyl- cells selecting for ampR xyl+ transformants . A 21- and a 27-kb plasmid were isolated, both of which contained the entire xylose regulon . The xylose regulon was localized to a 6.3-kb segment of a 13.5-kb EcoRI fragment . Subclones were constructed which contained either the genes for D-xylose isomerase and D-xylulokinase or the genes for the D-xylose transport and the D-xylose regulatory factors . The gene order determined by the subcloning experiments is consistent with that determined by genetic mapping . The spots corresponding to D-xylose isomerase and D-xylulokinase subunits were identified in two-dimensional gels of several xylose-induced strains . Each of them had a molecular weight of 45,000 and an isoelectric point of 6.2 +/- 0.1. Mol Gen Genet, 1984, 195(1-2), 256 - 9 Application of phage lambda technology to Salmonella typhimurium . Construction of a lambda-sensitive Salmonella strain; Harkki A et al.; We have previously constructed a novel strain of S . typhimurium carrying the E . coli lamB gene and shown that this strain adsorbs phage lambda and, for example, can be used for transposon mutagenesis with lambda vectors . In this study, we show that this strain can support the lytic growth of phage lambda nin derivatives, but not growth of wild-type lambda . However, lysogenization with lambda nin does not occur . Using this strain as starting material we took the construction one step further by introducing the E . coli nusA gene in a multicopy plasmid to this strain . We could show that this new Salmonella derivative can support both the lytic and lysogenic mode of growth of several different lambda derivatives . Using the same approach it should be possible to construct lambda-sensitive derivatives of other enteric bacteria thus rendering them more amenable to in vivo genetic manipulation. Environ Mutagen, 1984, 6(5), 757 - 62 Mutagenicity of lichen constituents; Shibamoto T et al.; Usnic acid (the most abundant lichen constituents), physodic, and physodalic acids isolated from Hypogymnia enteromorph (Ach.) Nyl . were tested for mutagenicity in the Ames Salmonella/microsome assay . Physodalic acid exhibited clear dose-related mutagenicity against Salmonella typhimurium strain TA 100 with or without S9 mix in both plate-incorporation and preincubation assays . The addition of S9 mix increased the number of revertants approximately threefold and fourfold in preincubation and plate-incorporation assays, respectively. Teratog Carcinog Mutagen, 1984, 4(3), 273 - 83 Mutagenic, cytotoxic, and teratogenic effects of 2-acetylaminofluorene and reactive metabolites in vitro; Faustman-Watts EM et al.; The embryotoxic, mutagenic, and cytotoxic properties of 2-acetylaminofluorene (AAF) and two of its reactive metabolites, N-acetoxy-2-acetylaminofluorene (AAAF) and 2-nitrosofluorene (NF) were assessed in vitro . A combined embryo culture/biotransformation system was used to determine the ability of these compounds to produce embryonic malformations, growth retardation, and/or embryolethality . Salmonella typhimurium auxotrophs (his-) were utilized to measure the mutagenic and cytotoxic potentials of these compounds . The parent compound, AAF, did not produce embryonic malformations or mutagenicity in the absence of an added cytochrome P-450-dependent monooxygenase system . Both metabolites produced each of the measured toxic effects without supplementation of a bioactivation system . However, the three chemicals each elicited a different spectrum of malformations . Bioactivated AAF produced neural tube abnormalities, whereas embryos treated with AAAF primarily exhibited prosencephalic malformations, and NF produced abnormalities of axial rotation or flexure . NF was approximately ten times more potent than AAAF as a direct-acting mutagen but only slightly more active in producing embryonic malformations in vitro . The results indicated that differential effects on the various measured parameters could be produced by these chemicals . The results indicated further that neither NF nor AAAF appeared to be individually responsible for the neural tube abnormalities generated by biotransformed AAF. Mol Gen Genet, 1984, 193(1), 135 - 42 Evidence that nitrogen regulatory gene ntrC of Salmonella typhimurium is transcribed from the glnA promoter as well as from a separate ntr promoter; Krajewska-Grynkiewicz K et al.; Previous work has indicated that nitrogen regulatory genes ntrB and ntrC of Salmonella typhimurium are closely linked to glnA, the structural gene encoding glutamine synthetase; proceeding clockwise the order of genes in the 86 U region of the map is polA...ntrC ntrB glnA glnA promoter...rha . To study ntrC transcription we have constructed operon fusions of ntrC to lacZ using the Casadaban Mu d1 (Apr lac) phage so that we can measure beta-galactosidase activity as a reflection of ntrC transcription and we have introduced into fusion strains promoter constitutive mutations at glnA {glnAp(Con)} . The glnAp(Con) mutations, which elevate glnA expression in fusion strains, also elevate beta-galactosidase activity, indicating that ntrC is cotranscribed with glnA . Consistent with this interpretation, polar insertion mutations in glnA decrease beta-galactosidase activity of fusion strains carrying glnAp(Con) mutations . However, glnA insertions do not eliminate beta-galactosidase activity of glnAp(Con) ntrC::Mu d1 strains and they have little effect on beta-galactosidase activity of the original ntrC::Mu d1 fusion strains . The latter results confirm that ntrC can also be transcribed from an ntr promoter downstream of glnA . Polar insertion mutations in ntrB eliminate beta-galactosidase activity of both the original fusion strains and fusion strains carrying glnA(Con) mutations, indicating that the ntr promoter lies between glnA and ntrB. IARC Sci Publ, 1984, (63), 59 - 88 Liver carcinogenesis in tropical Africa; Uwaifo AO et al.; The geographical pathology of hepatocellular carcinoma (HCC) was essentially anecdotal until systematic cancer registration was introduced, but it is now clear that sub-Saharan Africa is a high-incidence area . The disease is multifactorial in etiology, the possible etiological agents including hepatitis B virus and a number of chemical carcinogens, among which the most important appear to be the aflatoxins and N-nitroso compounds . Medicinal plants and herbal teas used in the tropics also contain compounds such as furocoumarins that are mutagenic and/or carcinogenic . A modified form of the Ames Salmonella typhimurium assay was used to study the mutagenicity of aflatoxins B1 and M1 and palmotoxin B0, a co-metabolite of aflatoxin B1, and also of chamuvaritin and chamuvarin, two benzyldihydrochalcones derived from the roots of Uvaria chamae, which are used for medicinal purposes in West Africa . The mutagenicity of a number of furocoumarins isolated from Nigerian medicinal plants was studied by means of the same assay as well as with Chinese hamster V79 cells and C3H 10T1/2 cells; in the last two systems the studies were carried out both with and without photoactivation . The same compounds, and 8-methoxypsoralen, were also investigated by means of cell transformation studies. Mol Gen Genet, 1984, 195(1-2), 219 - 27 Isolation and characterization of lac fusions to two nitrogen-regulated promoters; Stern MJ et al.; Mud1 (Ap, lac, cts)-mediated fusions to argTr and dhuA, two transport operon promoters in Salmonella typhimurium, were isolated and characterized in order to investigate the regulation of these promoters . Using these fusions we showed that these promoters are under nitrogen regulation and that this effect, as well as the response to a promoter-up mutation in dhuA, is at the transcriptional level . We utilized the fusions to determine that the histidine transport operon does not contain any internal promoters . The fusions were also used to screen the promoters for additional modes of regulation: argTr was found to respond to carbon regulation in addition to nitrogen regulation, while dhuA does not . The argTr promoter contains a sequence with good homology to the consensus sequence determined for the cAMP receptor protein binding site . Neither promoter responds to sulfur or phosphate regulation. Biochemistry, 1983 Dec 20, 22(26), 6130 - 4 Phosphorus-31 nuclear magnetic resonance investigation of membrane vesicles from Escherichia coli; Hunt AG et al.; Phosphorus-31 nuclear magnetic resonance studies of isolated membrane vesicles prepared from Escherichia coli PSM116 as described by Hunt and Hong {Hunt, A . G., & Hong, J.-S . (1981) J . Biol . Chem . 256, 11988-11991; Hunt, A . G., & Hong, J.-S . (1983) Biochemistry 22, 844-850} are detailed here . This strain harbored a recombinant plasmid containing the phosphoglycerate transport system from Salmonella typhimurium (pJH7) . Evidence indicating a surprising metabolic diversity, such as the presence of the enzymes enolase and phosphoglycerate mutase, is presented . The nature of the energization of these membrane vesicles for transport as described by Hugenholtz et al . {Hugenholtz, J., Hong, J.-S., & Kaback, H . R . (1981) Proc . Natl . Acad . Sci . U.S.A . 78, 3446-3449} is also discussed . Membrane vesicles prepared from the PSM116 strain do not form a transmembrane pH gradient when phosphoenolpyruvate is added . The present results show that phosphorus-31 nuclear magnetic resonance spectroscopy is an excellent tool to investigate the metabolism of membrane vesicles. Pharm Weekbl Sci, 1983 Dec 16, 5(6), 302 - 7 Correlations between phototoxicity of some 7-chloro-1,4-benzodiazepines and their (photo)chemical properties; De Vries H et al.; In relation to the phototoxicity of 7-chloro-I,4-benzodiazepine-N-oxides the photostability of some of these N-oxides and the thermostability of their photoproducts, the oxaziridines, were studied . Rather than a consequence of a direct phototoxic effect by the N-oxides the ultimate toxic effect in the test system Salmonella typhimurium appeared to be caused by products formed during and after the irradiation . For chlordiazepoxide (CDZ) and its main metabolites in man desmethylchlordiazepoxide (DES) and demoxepam (DEM) the formation of an oxaziridine is indeed a crucial factor for the onset of the toxic effect . However, DEM oxaziridine (DEM OX.) being very thermolabile in protic medium forms non-toxic conjugates with the solvent . CDZ OX . and DES OX . are thermally converted into their N-oxide, although DES OX . partly decomposes into 6-chloro-4-phenylquinazoline-2-carboxaldehyde as well . This compound proved to be an important factor in the toxic action of DES after irradiation. Biochem Pharmacol, 1983 Dec 15, 32(24), 3791 - 5 Relative mutagenicity and teratogenicity of cyclophosphamide and two of its structural analogs; Hales BF; In this report, cyclophosphamide was compared to two of its structural analogs, 5,5-dimethylcyclophosphamide and diethylcyclophosphamide, with respect to mutagenic and teratogenic activities . Mutagenicity was assessed using Salmonella typhimurium TA 1535; teratogenicity was assessed in Sprague-Dawley rats on day 20 of gestation after intra-amniotic drug administration on day 13 . After metabolic activation, cyclophosphamide caused base substitution mutations in S . typhimurium TA 1535 and major structural defects in both intra-amniotically injected and contralateral uninjected fetuses . 5,5-Dimethylcyclophosphamide was neither mutagenic nor teratogenic . Diethylcyclophosphamide was not mutagenic but was teratogenic . However, diethylcyclophosphamide was less potent as a teratogen than cyclophosphamide and, unlike cyclophosphamide, caused malformations only in the intra-amniotically injected fetuses . Diethylcyclophosphamide does liberate acrolein after metabolic activation . If acrolein is responsible for the teratogenic effects of diethylcyclophosphamide, the other major cytotoxic metabolite of cyclophosphamide, phosphoramide mustard, may account for the difference in teratogenic potency between cyclophosphamide and diethylcyclophosphamide . These results would suggest that acrolein, although apparently not mutagenic, mediates the teratogenicity of diethylcyclophosphamide and a significant proportion of the teratogenicity of cyclophosphamide. Biochem Pharmacol, 1983 Dec 15, 32(24), 3755 - 63 Genetic expression of aflatoxin metabolism . Effects of 3-methylcholanthrene and beta-naphthoflavone on hepatic microsomal metabolism and mutagenic activation of aflatoxins; Raina V et al.; The effects of pretreatment with 3-methylcholanthrene (MC) and beta-naphthoflavone (beta NF) on the hepatic microsome-mediated mutagenesis of aflatoxin B1 (AFB1) and benzo{a}pyrene, and on the metabolism of aflatoxins B1 and B2, were investigated in inbred mouse strains . The inbred strains of mice studied included Ah nonresponsive strains (DBA/2Ha, AKR/Sn and RF/J), which were also nonresponsive to the induction of the metabolism of AFB1 to AFM1 (AFB1-4-hydroxylase activity), and Ah responsive strains (C57BL/6Ha, ICR/Ha, C3H/St, A/St, Balb/cCr, C57e/Ha and CBA/Pi), which were also responsive to the induction of AFB1-4-hydroxylase activity . The hepatic microsome-mediated enzyme activities studied included: mutagenic activation of AFB1 and benzo{a}pyrene in the Ames Salmonella typhimurium TA-98 system; metabolism of AFB1 and AFB2 to AFM1 and AFM2, respectively; and benzo{a}pyrene metabolism measured as the formation of fluorescent phenolic metabolites, i.e . aryl hydrocarbon hydroxylase (AHH) activity . Time-course and dose-response studies in C57BL/6Ha mice revealed that the metabolism of aflatoxin B1/B2 to aflatoxin M1/M2 (AFB1/B2-4-hydroxylase activity) was induced by both MC and beta NF . In the nonresponsive strains studied, pretreatment with MC or beta NF produced essentially little alteration of AFB1-4-hydroxylase activity or AHH activity or the mutagenic activation of AFB1 and benzo{a}pyrene . On the other hand, AFB1-4-hydroxylase activity in the responsive strains was induced 4- to 10-fold by MC (60 mg/kg) and 2.5- to 7-fold by beta NF (150 mg/kg) . Also in the responsive strains, induction of AFB1-4-hydroxylase activity was strongly associated with (a) the depression of the mutagenic activation of AFB1, and (b) with the induction of both AHH and the mutagenic activation of benzo{a}pyrene . In summary, the results described in this report suggest that: (a) induction of AFB1-4-hydroxylase activity by MC (or beta NF) is associated with the depression of AFB1 mutagenesis and with the induction of benzo{a}pyrene mutagenesis; and (b) induction by MC (or beta NF) of AHH activity, AFB1-4-hydroxylase activity and AFB2-4-hydroxylase activity is controlled by either the same or closely linked genetic factors. Food Chem Toxicol, 1983 Dec, 21(6), 815 - 23 Safety evaluation of thaumatin (Talin protein); Higginbotham JD et al.; Thaumatin, the sweet proteinaceous extract of the arils of Thaumatococcus daniellii (Benth.) has been studied for its subacute toxicity in rats and dogs and its ability to produce anaphylactic antibodies following oral administration to rats and normal human subjects . Thaumatin was readily digested prior to absorption in rats and no adverse effects resulted from its continuous administration to rats and dogs at dietary concentrations of 0, 0.3, 1.0 and 3.0% for 13 wk . It was not teratogenic when administered orally to rats at 0, 200, 600 and 2000 mg/kg body weight/day from day 6 to 15 of gestation and was without effect on the incidence of dominant lethal mutations when administered on five consecutive days to male mice at 200 and 2000 mg/kg/day . The lack of mutagenic potential was confirmed in bacterial mutagenic assays with Salmonella typhimurium (strains TA1535, TA1537, TA1538, TA98 and TA100) and Escherichia coli WP2, at levels of addition of 0.05-50 mg/plate . In rats, thaumatin was found to be a weak sensitizer, comparable with egg albumen, when administered systemically but to be inactive when administered orally . Prick testing of laboratory personnel who had been intermittently exposed by inhalation to thaumatin for periods up to 7 yr showed that 9.3% (13/140) responded positively to commercial thaumatin, while 30.7% were positive to Dermatophagoides pteronyssinus (house dust mite) . None of the subjects who gave a positive skin reaction to commercial thaumatin responded to the plant components remaining after removal of the specific sweet Thaumatin proteins . Challenge tests in man did not demonstrate any oral sensitization . The results indicate that thaumatin when used as a flavour modifier and extender, and partial sweetener, is unlikely to be hazardous at the anticipated level of consumption. Mutat Res, 1983 Dec, 124(3-4), 213 - 24 Comparative genotoxicity studies of the flame retardant tris(2,3-dibromopropyl)phosphate and possible metabolites; Holme JA et al.; Tris(2,3-dibromopropyl)phosphate (Tris-BP) was activated to mutagens in the Salmonella/microsome quantitative test system . Liver microsomes from rats pretreated with phenobarbital (PB) increased the mutagenicity of 0.05 mM Tris-BP to 186% of the activity obtained with liver microsomes from untreated rats . The addition of 0.02 mM Tris-BP to V79 Chinese hamster cells co-incubated with liver microsomes from PB-pretreated rats increased the number of mutants by a factor of 9.7 . Tris-BP also caused genotoxic and cytotoxic responses in primary monolayers of rat hepatocytes . The relative increase in unscheduled DNA synthesis after treatment with 0.05 mM Tris-BP was 2.3-fold as measured by scintillation counting of radiolabelled thymidine incorporated into DNA of isolated nuclei . The use of hepatocytes isolated from PB-pretreated rats reduced the increases in DNA repair synthesis relatively to that in control cells . Monolayers of hepatocytes from untreated rats co-cultured with Salmonella typhimurium TA100 activated Tris-BP to mutagenic intermediates which were released into the culture medium . The studies with the V79 and liver-cell systems indicate that the reactive intermediates formed from Tris-BP are sufficiently stable and lipophilic to traverse the various membranes from the site of generation to the respective cellular targets . The relative degree of genotoxic responses of bis(2,3-dibromopropyl)phosphate, 2,3-dibromopropylphosphate, tris(2,3-bromopropyl)phosphate, tris(2-bromopropyl)phosphate and 2,3-dibromopropanol in the systems studied did not indicate that these compounds were proximate or ultimate reactive metabolites of Tris-BP in liver-derived activation systems. J Med Chem, 1983 Dec, 26(12), 1715 - 9 Heterocyclic Quinones . 4 . A new highly cytotoxic drug: 6,7-bis(1-aziridinyl)-5,8-quinazolinedione; Renault J et al.; With the aim of obtaining new antitumoral agents, a series of 5,8-quinazolinediones was prepared . 5-Amino-6-methoxyquinazoline was oxidized by Fremy's salt to give 6-methoxy-5,8-quinazolinedione . Nucleophilic substitution reaction at C6, electrophilic substitution at C7, and synthesis of 7-amino-6-methoxy-5,8-quinazolinedione, the parent compound of streptonigrin, were studied . These compounds were tested for cytotoxic properties on L1210 leukemia cells in vitro . One of them, 6,7-bis(1-aziridinyl)-5,8-quinazolinedione, which exhibits a high cytotoxic activity (ID50 = 0.08 microM), was further screened in standard antitumor systems, including L1210 leukemia, P388 lymphocytic leukemia, sarcoma 180, and B16 melanocarcinoma . This drug gives a significant antitumoral effect on P388 leukemia but is inactive on other experimental models . Moreover, this compound was found to be highly mutagenic for Salmonella typhimurium TA98 and TA100 strains (Ames test), suggesting that DNA damage could be responsible for its cytotoxicity. Cancer Res, 1983 Dec, 43(12 Pt 1), 5768 - 74 Microsomal activation of 2-amino-3-methylimidazo{4,5-f}quinoline, a pyrolysate of sardine and beef extracts, to a mutagenic intermediate; Yamazoe Y et al.; The mechanism involved in the metabolic activation of 2-amino-3-methylimidazo{4,5-f}quinoline, which is a pyrolysate isolated from broiled foods, to a mutagenic intermediate was studied in vitro . In a system containing hepatic microsomes and reduced nicotinamide adenine dinucleotide phosphate, 2-amino-3-methylimidazo{4,5-f}quinoline was converted to a product which was directly mutagenic to Salmonella typhimurium . The structure of the mutagenic metabolite was determined as the 2-N-hydroxy derivative on the basis of the chemical properties and the mass spectral evidence of the azoxy adduct with o-nitrosotoluene . The activation reaction was mediated by microsomal enzymes and was inhibited by carbon monoxide, 7,8-benzoflavone, and other chemicals which were known to inhibit the cytochrome P-450-dependent reaction . With the use of four forms of purified cytochrome P-450, the N-hydroxylation of 2-amino-3-methylimidazo{4,5-f}quinoline and the induction of the reverse mutation of the bacteria were clearly demonstrated to be catalyzed mainly by a high-spin form of cytochrome P-450, P-448 II-a. J Appl Toxicol, 1983 Dec, 3(6), 317 - 20 The genetic activity of anthramycin, tomaymycin and sibiromycin in bacterial forward- and reverse-mutation assays and in the mouse bone-marrow micronucleus test; Gairola C et al.; The genetic activity of the structurally similar antitumor antibiotics anthramycin, tomaymycin and sibiromycin was evaluated in the standard Ames Salmonella/microsome mutagenicity assay, a Salmonella typhimurium forward-mutation assay and the micronucleus test . None of the test drugs showed any significant genetic activity in forward or reverse Salmonella mutation assays . The ability of mouse-liver enzymes to produce mutagens from the drugs was examined in the Salmonella reverse-mutation assay and was generally negative . As the concentrations of sibiromycin increased, some activity was detected in the presence of liver S-9 fractions from Aroclor-induced mice . This observation could not be verified at higher concentrations in the reverse-mutation assay due to cytotoxicity, and in the forward-mutation assay due to interference with the selection process by S-9 . Cytogenetic evaluation of anthramycin and tomaymycin in the micronucleus test also gave negative results . However, significant increases in the frequency of micronucleated polychromatic erythrocytes were observed in the bone marrow of sibiromycin-treated mice . The results suggest that, except for some possible activity of sibiromycin, these drugs are generally devoid of any marked genetic activity in the test systems employed. Bangladesh Med Res Counc Bull, 1983 Dec, 9(2), 37 - 42 Effect of vitamin-A deficiency on plaque forming response of antibody producing spleen cells against Salmonella typhimurium in rats; Faruque SM et al.; Plaque forming response of antibody producing spleen cells against Salmonella typhimurium was studied in vitamin-A deficient and normal rats after 3, 6, 9 and 12 days of injecting the antigen . Vitamin-A deficient rats were found to have significantly decreased (P less than 0.001) number of antibody plaque forming cells in the spleen as compared to normal rats in all cases . Serum total protein and serum Vitamin-A levels were significantly (P less than 0.001) lower in the vitamin-A deficient rats as compared to the controls and immunization caused no significant change in these parameters . The average spleen weights were increased in both the groups on immunization but this increase was comparatively more in case of the control rats. Proc Natl Acad Sci U S A, 1983 Dec, 80(24), 7496 - 500 AppppA, heat-shock stress, and cell oxidation; Lee PC et al.; Salmonella typhimurium LT2 induces a set of heat-shock proteins analogous to those found previously in Escherichia coli . These are virtually the only proteins synthesized after a temperature shift from 28 degrees C to 50 degrees C . Using a two-dimensional thin-layer chromatographic system developed to resolve adenylylated nucleotides, we have found that S . typhimurium and E . coli accumulate P1,P4-diadenosine-5'-tetraphosphate (AppppA), P1-(adenosine-5')-P3-(guanosine-3'-diphosphate-5')-triphosphate (ApppGpp), P1-(adenosine-5')-P4-(guanosine-5')-tetraphosphate (AppppG), P1-(adenosine-5')-P3-(guanosine-5')-triphosphate (ApppG), and P1,P3-diadenosine-5'-triphosphate (ApppA) after heat shock . These same adenylylated nucleotides accumulate after exposure to ethanol, an agent also known to induce the heat-shock response in a variety of cells . AppppA, ApppGpp, AppppG, ApppG, and ApppA were previously shown to accumulate under conditions of oxidation stress . We proposed that these adenylylated nucleotides may be alarmones--i.e., regulatory molecules, alerting cells to the onset of oxidation stress . The finding that these dinucleotides accumulate in response to heat shock suggests that oxidation and heat shock have a common physiological effect on cells . We hypothesize that these dinucleotides signal the onset of these stresses and trigger the "heat-shock response." Diagn Microbiol Infect Dis, 1983 Dec, 1(4), 335 - 7 A pseudoepidemic due to Salmonella typhimurium; Harris AA et al.; A pseudoepidemic due to Salmonella typhimurium occurred in the clinical microbiology laboratory of a university hospital and involved 10 patients . One patient received "unnecessary" antibiotics . Investigation of the events implicated a contaminated rubber pipette bulb . Such bulbs should be considered as a possible source of false-positive cultures. Clin Physiol, 1983 Dec, 3(6), 551 - 63 Biochemical responses of the myocardium and red skeletal muscle to Salmonella typhimurium infection in the rat; Ilback NG et al.; Previous studies with bacterial infections have demonstrated a reduced exercise capacity and equally pronounced catabolic responses in red and white skeletal muscle . In the present study, red skeletal muscle and heart ventricular muscle were compared in a S . typhimurium model in rats . Two days before median lethality was achieved, the activities of one oxidative (cytochrome c oxidase), one glycolytic (glyceraldehyde-3-phosphate dehydrogenase) and one lysosomal (beta-glucuronidase) enzyme were determined in the two tissues . The contents of protein, RNA and DNA were also determined . The oxidative and glycolytic capacity decreased 24-29% in red skeletal muscle but only 7-20% in the myocardium . However, the decrease in oxidative capacity in skeletal muscle and myocardium was statistically correlated . The protein synthetic capacity (RNA) also decreased and was correlated to the protein concentration in both tissues . This metabolic impairment of both skeletal and heart muscle probably contributes to the deterioration of the physical performance capacity previously observed to follow acute infectious diseases . This study emphasizes the importance of the choice of reference, such as 'wet' weight, DNA or the entire organ, when evaluating metabolic results in biologic tissues and that biochemical alterations in skeletal muscle biopsies in bacterial infections do not reflect alterations in myocardium reliably. Food Chem Toxicol, 1983 Dec, 21(6), 745 - 7 Possible mutagenic constituents in nitrite-treated soy sauce; Shibamoto T; Soy sauce was heated with 100, 500, 1000 or 2000 ppm sodium nitrite for 30 min at 80 degrees C and pH 3 . The reaction mixtures were extracted with dichloromethane followed by ethyl acetate . After removal of the solvents, the extracts were subjected to analysis (gas chromatograph-thermal energy analyser and gas chromatograph-mass spectrometer) and Ames mutagenicity tests . N-Nitrosodimethylamine and N-nitrosodiethylamine were found in the dichloromethane extract of the soy sauce treated with 2000 ppm nitrite at levels of 10 and 120 micrograms/ml, respectively . N-Nitrosoproline was identified in the ethyl acetate extract of the same sample at a level of 0.5 microgram/ml . Both extracts exhibited dose-related mutagenicity in Salmonella typhimurium strain TA100 with S-9 mix . The dichloromethane extract showed much higher mutagenicity than did the ethyl acetate extract . The samples obtained from soy sauce treated with 100, 500 and 1000 ppm nitrite were not mutagenic, but N-nitrosodiethylamine was detected by thermal energy analysis in the soy sauce treated with 1000 ppm nitrite . The addition of 10,000 ppm L-ascorbic acid, along with 2000 ppm nitrite, to soy sauce prevented the formation of mutagenic materials or detectable nitrosamines. Mutat Res, 1983 Dec, 124(3-4), 315 - 24 Bioactivation and biotransformation of 1-nitropyrene in liver, lung and nasal tissue of rats; Bond JA; 1-Nitropyrene (NP) is a known direct-acting bacterial mutagen and has been detected in the environment from such sources as diesel-exhaust emissions and coal-combustion fly ash . The purpose of this study was to investigate the mutagenic potential of NP in Salmonella typhimurium using rat liver, lung and nasal tissue as the enzyme-activating systems and to measure the rates of NP metabolism in these same tissues . Rat liver, lung and nasal tissue bioactivated NP to mutagens that were detected in the Ames bacterial test system . At all doses of NP and all protein concentrations of tissue S9, mutagenic responses were larger than that observed in the absence of any tissue . In both strains TA98 and TA100, about 1.0 mg/ml liver and nasal tissue S9 appeared to be the optimal concentration which resulted in the largest mutagenic response to NP, whereas 2.0 mg/ml of lung S9 was necessary to yield optimal responses . When NP was incubated with liver, lung or nasal tissue S9 and strain TA98 NR, mutagenic responses were significantly decreased when compared to the response seen in TA98 . NP was metabolized to several oxidized metabolites (3-, 6- and 8-hydroxynitropyrene) in all tissues examined . Total rates of formation of NP metabolites for nasal tissue, liver and lung S9 were 650, 300 and 60 pmoles/mg protein/min, respectively . These results suggest that the respiratory tract, in particular the nasal tissue, may be an important site for in vivo bioactivation of inhaled NP. Mutat Res, 1983 Dec, 122(3-4), 279 - 86 Formation of mutagens by amino-carbonyl reactions; Shinohara K et al.; The formation of mutagens by amino-carbonyl reactions of 20 kinds of amino acid and sugars after heating at 100 degrees C for 10 h was examined by the Ames test . The browned solutions of Gly, Ala, Val, Leu, Ile, Ser, Thr, Gln, Lys X HCl, Arg, Phe, Cys, Met and Pro with Glc caused mutation of Salmonella typhimurium TA100 and/or TA98 with or without S9 mix . The presence of S9 mix increased the mutagenic activity of the browned solutions of Cys and Phe with Glc on TA100 and of those of Gly, Ala, Val, Ile and Cys on TA98, but decreased the activity of other solutions . No revertants of Salmonella were induced by the browned solutions of Trp, Tyr, Asp, Asn, Glu and (Cys)2 with Glc . Among positive browned solutions, Cys, Lys, Arg and Phe had the stronger activity, but their activity was weak compared with that of pyrolysates or chemical mutagens such as Trp-P-1, Trp-P-2 and 4-nitroquinoline-N-oxide . The mutagenic activity of the browned solutions increased with prolongation of heating time and varied with the pH of the reaction mixture . Fru, Gal, Ara, Xyl, Man, Lac and Suc also had the ability to form mutagens in the browning reactions with amino acids. Mutat Res, 1983 Dec, 122(3-4), 273 - 8 Mutagenicity of coal-pyrolyzed products and their photochemical reaction products with nitrogen oxides in Salmonella typhimurium TA98 and TA100; Hirayama T et al.; The chemical class separation of coal-pyrolyzed products and the photochemical reaction of these fractions with nitrogen oxides in the experimental chamber, and the application of a short-term mutagenicity test were investigated . The altered products from the fraction hydroxy polycyclic aromatic compounds in a simulated atmosphere containing a small volume of nitrogen oxides under irradiation with a xenon lamp were the most potent mutagenic fraction among all the fractions tested against Salmonella typhimurium, both TA98 and TA100, with or without S9. Mutat Res, 1983 Dec, 122(3-4), 267 - 72 Genetic damage in Salmonella typhimurium by near-ultraviolet radiation . Lack of repair by plasmid pKM101; Eisenstark A; Plasmid pKM101, whose mucA and B genes endow cells with enhanced mutation frequency and enhanced resistance to far-ultraviolet radiation (FUV) (254 nm), had no influence on these properties when cells were damaged by near-ultraviolet radiation (NUV) (300-400 nm) . Thus, NUV lesions did not lead to induction of SOS repair and subsequent expression of mucA and B genes on plasmid pKM101 . Further, when cells were pre-irradiated with NUV and subsequently irradiated with FUV, there was a blockage of SOS repair, including the repair normally controlled by genes on pKM101. Mutat Res, 1983 Dec, 122(3-4), 257 - 66 Isolation of a mutant of Salmonella typhimurium strain TA1535 with decreased levels of glutathione (GSH-) . Primary characterization and chemical mutagenesis studies; Kerklaan P et al.; A mutant of Salmonella typhimurium strain TA1535 with decreased glutathione (GSH) levels was isolated after treatment with UV and selection for N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) resistance; this GSH- mutant also exhibited increased resistance to MNNG, the methyl analog of ENNG . Estimation of the cellular GSH content showed that the GSH- derivative contained about 20% of the GSH levels found in TA1535 . In mutagenicity tests (hisG46 leads to His+), the GSH- strain required the presence of GSH or L-cysteine in the medium for an optimal phenotypic expression and/or growth of spontaneous and induced His+ revertants, and may, therefore, be allelic to cys mutants of Salmonella described earlier . The mutagenic activity of MNNG, ENNG and 1,2-dibromoethane (DBE), but not that of N-ethylnitrosourea (ENU), was strongly reduced in TA1535/GSH-; pretreatment of the strain with GSH restored the mutagenicity of the first 3 chemicals to levels normally found in TA1535 . The results support the current view that MNNG, ENNG and DBE, but not ENU, can be activated via reaction with GSH to species of higher reactivity and mutagenicity . It is concluded that the present GSH- strain can be used to study more systematically the role of GSH in the bioactivation and -deactivation of xenobiotics to mutagenic factors. Carcinogenesis, 1983 Dec, 4(12), 1615 - 8 The synthesis and mutagenicity of the N-formyl analog of N-hydroxyphenacetin; Corbett MD et al.; The synthesis and purification of N-hydroxy-N-formyl-p-phenetidine (N-OH-FP) is described . This new compound was subjected to mutagenicity testing using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 both in the presence and absence of the post-mitochondrial fraction of rat liver homogenate . Simultaneous mutagenicity testing of the known phenacetin metabolite, N-hydroxyphenacetin (N-OH-AP), was conducted with the same tester strains . The N-formyl derived hydroxamic acid (N-OH-FP) was found to be a much stronger mutagen than N-hydroxy-phenacetin (N-OH-AP) . Furthermore, N-OH-FP also behaved as a direct-acting mutagen unlike N-OH-AP . The chemical stabilities of N-OH-AP and N-OH-FP were studied in phosphate buffer in the pH range of 3-8; and both the hydroxamic acids were found to be stable to the conditions employed . The results of this study support the hypothesis that enzymatic deacylation is an activation process for the expression of mutagenicity by hydroxamic acids. Carcinogenesis, 1983 Dec, 4(12), 1551 - 7 Activation and detoxication of N-hydroxy-Trp-P-2 by glutathione and glutathione transferases; Saito K et al.; The roles of non-enzymatic and enzymatic glutathione (GSH) conjugation in the activation and detoxication of 3-hydroxy-amino-1-methyl-5H-pyrido{4,3-b}indole (N-OH-Trp-P-2) were studied in vitro . N-OH-Trp-P-2 is an active metabolite of 3-amino-1-methyl-5H-pyrido{4,3-d}indole (Trp-P-2), a mutagenic and carcinogenic heterocyclic amine . 3-Nitroso-1-methyl-5H-pyrido{4,3-b}indole (NO-Trp-P-2) reacted rapidly and non-enzymatically with GSH to form N-OH-Trp-P-2 and a small amount of two GSH conjugates (CN-1 and CN-2) . On the other hand, non-enzymatic reaction of GSH with N-OH-Trp-P-2 was very slow, but the GSH conjugation with N-OH-Trp-P-2 was catalyzed by rat liver GSH transferase and a rat liver cytosol fraction to form three conjugates (CH-1, CH-2 and CH-3) . The enzymatic conjugation was effectively inhibited by organic tin compounds which are known as powerful GSH transferase inhibitors . The conjugates were unstable enough to yield Trp-P-2 (from CN-1, CN-2 and CH-2) or N-OH-Trp-P-2 (from CH-3) on incubation at 37 degrees C for 30-60 min . Only CH-1 was stable under similar conditions . The mutagenicities of the GSH conjugates and the effects of GSH and GSH transferase were studied by using Salmonella typhimurium TA98 as the tester strain . The GSH conjugates except for CH-3 were completely detoxicated products, but CH-3 was found to be a more potent mutagen than N-OH-Trp-P-2 . The mutagenicity of CH-3 seemed to be due to the direct action of the conjugate, and not to N-OH-Trp-P-2 formed from it. Carcinogenesis, 1983 Dec, 4(12), 1547 - 50 Syntheses of hydroxyamino, nitroso and nitro derivatives of Trp-P-2 and Glu-P-1, amino acid pyrolysate mutagens, and their direct mutagenicities towards Salmonella typhimurium TA98 and TA98NR; Saito K et al.; Hydroxyamino, nitroso and nitro derivatives of 3-amino-1-methyl-5H-pyrido{4,3-b}indole (Trp-P-2) and 2-amino-6-methyldipyrido{1,2-a:3',2'-d}imidazole (Glu-P-1), mutagens-carcinogens produced on pyrolysis of amino acids, were synthesized from Trp-P-2 and Glu-P-1 . 3-Hydroxyamino-1-methyl-5H-pyrido{4,3-b}indole (N-OH-Trp-P-2) and 2-hydroxyamino-6-methyldipyrido{1,2-a:3',2'-d}imidazole (N-OH-Glu-P-1) were obtained with good yields by controlled catalytic reduction of 3-nitro-1-methyl-5H-pyrido{4,3-b}indole and 2-nitro-6-methyldipyrido{1,2-a:3',2'-d}imidazole . Subsequent oxidation of N-OH-Trp-P-2 and N-OH-Glu-P-1 with gamma-manganese dioxide yielded 3-nitroso-1-methyl-5H-pyrido{4,3-b}indole and 2-nitroso-6-methyldipyrido{1,2-a:3',2'-d}imidazole . All six synthesized compounds were mutagenic to Salmonella typhimurium TA98 without mammalian activation systems . The mutagenic activities of hydroxyamino and nitroso derivatives were identical for both S . typhimurium TA98 and TA98NR, the nitroreductase deficient strain . However, nitro derivatives were essentially mutagenic only towards S . typhimurium TA98. Proc Natl Acad Sci U S A, 1983 Dec, 80(23), 7142 - 5 Kinetic characterization and regulation of phosphoenolpyruvate-dependent methyl alpha-D-glucopyranoside transport by Salmonella typhimurium membrane vesicles; Liu KD et al.; Membrane vesicles from Salmonella typhimurium SB3507 were used to study the kinetics of methyl alpha-D-glucopyranoside (MeGlc) transport by the phosphoenolpyruvate: glycose phosphotransferase system (PTS) . During the first minute of phosphoenolpyruvate-dependent MeGlc transport, two distinct rates were observed; an initial rapid rate, V1 (Vmax, 7.4-8.4 nmol X mg-1 X min-1; Km, 8.2-11.2 X 10(-6)M), followed by a second slower rate, V2 (Vmax, 4-4.6 nmol X mg-1 X min-1; Km, 3.4-6.4 X 10(-6) M) . The change in rate occurred when the intravesicular MeGlc phosphate concentration was 0.2 mM or less, depending on the external MeGlc concentration . The rate-limiting component in MeGlc transport was found to be enzyme II-BGlc, not phosphoenolpyruvate uptake or the PTS proteins enzyme I, HPr, and IIIGlc . The change from V1 to V2 thus suggests that the PTS is regulated in intact vesicles . However, this regulation was completely relieved by permeabilizing the vesicles with toluene . That is, the toluene-treated vesicles showed only V1 for MeGlc phosphorylation . Evidence was obtained to show that pyruvate and its metabolic products generated by the vesicles exerted no effect on the rate of MeGlc transport . Furthermore, the result from a dual-label experiment excluded exchange transphosphorylation as the mechanism for regulating MeGlc transport by the vesicles . Possible mechanisms for regulation of the PTS are discussed. J Immunol, 1983 Dec, 131(6), 3006 - 13 Genetic control of the innate resistance of mice to Salmonella typhimurium: expression of the Ity gene in peritoneal and splenic macrophages isolated in vitro; Lissner CR et al.; The mouse Chromosome 1 locus Ity regulates the extent to which Salmonella typhimurium replicates within the reticuloendothelial cell system (RES) during the first days of infection . If animals are homozygous for the Itys susceptibility allele, the Gram-negative bacterium undergoes rapid net multiplication, and mice die of a typhoid fever-like disease by day 10 of infection . Animals that are homozygous or heterozygous for the resistance allele, Ityr, control net bacterial replication and survive the first phase of salmonellosis . Indirect studies have implicated the resident macrophage as the effector cell for regulation of early in vivo salmonellae growth . To verify this supposition and to evaluate the phenotypic expression of Ity, we developed an in vitro assay to compare kinetics of S . typhimurium growth within Ityr and Itys macrophages . Resident peritoneal and splenic macrophages were used from inbred Ityr and Itys mice and from Ity congeneic mice . With these mice and through the use of radiolabeled S . typhimurium and an avirulent temperature-sensitive mutant of the bacterium, we found that: phagocytosis of S . typhimurium by Ityr and by Itys macrophages was the same; S . typhimurium grew to a greater extent in Itys peritoneal and splenic macrophages than in Ityr cells; Ityr macrophages killed intracellular salmonellae more efficiently than did Itys macrophages . Thus, we have demonstrated directly that Ity is expressed by the macrophage and have shown for the first time with Ity congeneic mice that the basis for differential net growth of virulent S . typhimurium in Ityr and Itys macrophages is a variation in the degree of bacterial kill. Infect Immun, 1983 Dec, 42(3), 1198 - 202 Effect of human leukocyte interferon on invasiveness of Salmonella species in HEp-2 cell cultures; Bukholm G et al.; The effect of human leukocyte interferon on the invasiveness of Salmonella and Shigella species in HEp-2 cell cultures was examined . The intracellular and extracellular bacteria were identified by a combination of Nomarski differential interference contrast microscopy and UV incident light microscopy applied on the same microscope . Pretreatment of HEp-2 cells with human leukocyte interferon reduced the number of Salmonella typhimurium and Salmonella paratyphi-B bacteria per cell and the proportion of cells containing bacteria in a dose-dependent manner . Maximum inhibitory effect was observed with ca . 100 U of interferon per ml . The inhibitory effect was neutralized with anti-human interferon globulin . Murine fibroblast interferon did not influence the invasiveness of Salmonella species . Invasiveness of Shigella flexneri was not influenced by treatment of cells with human interferon. Cancer Res, 1983 Dec, 43(12 Pt 1), 5821 - 5 Mutagenicity of the enantiomers of the diastereomeric bay-region benz(a)anthracene 3,4-diol-1,2-epoxides in bacterial and mammalian cells; Wood AW et al.; Enantiomers of the diastereomeric pair of bay-region benz(a)anthracene 3,4-diol-1,2-epoxides in which the benzylic 4-hydroxyl group and epoxide oxygen are either cis (isomer 1) or trans (isomer 2) were evaluated for mutagenic activity in two histidine-dependent strains of Salmonella typhimurium, as well as in an 8-azaguanine-sensitive Chinese hamster cell line . In strain TA 98 of S . typhimurium, the diol-epoxide with (1S,2R,3R,4S) absolute configuration {(-)-diol-epoxide 2} was the most active isomer, although there was less than a 3-fold difference in the mutagenicity of the four diol-epoxides . However, in strain TA 100 of S . typhimurium, the enantiomeric diol-epoxide with (1R,2S,3S,4R) absolute configuration {(+)-diol-epoxide 2} was the most active diol-epoxide, and the two isomers with (3S,4R) absolute configuration {(-)-diol-epoxide 1 and (+)-diol-epoxide 2} were three to eight times more active than were the two isomers with (3R,4S) configuration . The highest degree of sensitivity to absolute configuration was observed in Chinese hamster V79 cells, in which the (1R,2S,3S,4R) isomer {(+)-diol-epoxide 2} was from three to 20 times more mutagenic than were the other three isomers . This metabolically predominant (+)-diol-epoxide 2 isomer, which has high activity in strain TA 100 of S . typhimurium and the Chinese hamster V79 cells, has the same absolute configuration as do the bay-region diol-epoxide isomers of benzo(a)pyrene and chrysene that have been shown previously to be exceptionally mutagenic to mammalian cells and highly tumorigenic in mice . Analysis of the mutagenic activity of the (+)- and (-)-isomers of the 1,2- and 3,4-tetrahydroepoxides of benz(a)anthracene revealed only small enantiomeric differences in strain TA 98 of S . typhimurium (2.5 fold) and little, if any, differences (less than 1.5-fold) in the other two mutagenicity systems . However, the extent to which the four tetrahydroepoxides were converted to nonmutagenic products by homogeneous microsomal epoxide hydrolase (EC 3.3.2.3) indicated marked differences in the stereoselectivity of the enzyme . (-)-(3R,4S)-Epoxy-1,2,3,4-tetrahydrobenz(a)anthracene appears to be an exceptionally good substrate for epoxide hydrolase. Cancer Res, 1983 Dec, 43(12 Pt 1), 5713 - 7 Inactivation of a diol-epoxide and a K-region epoxide with high efficiency by glutathione transferase X; Glatt H et al.; Four glutathione transferases (EC 2.5.1.18), glutathione transferases A, B, and C and a hitherto unknown form, termed X, were purified to apparent homogeneity from rat liver cytosol . They were investigated for their abilities to inactivate two mutagenic epoxides derived from the polycyclic aromatic hydrocarbon benz(a)anthracene, the K-region epoxide benz(a)anthracene 5,6-oxide and the diol-epoxide r-8,t-9-dihydroxy-t-10,11-oxy-8,9,10, 11-tetrahydrobenz(a)anthracene . Mutagenic activity was determined using Salmonella typhimurium his- strain TA100 . Glutathione alone had little if any influence on the mutagenicity of the diol-epoxide but significantly decreased the mutagenic effect of the K-region epoxide . This inactivation was enhanced by the addition of glutathione transferases . Both epoxides were inactivated by glutathione in the presence of each of the four enzymes, but with varying efficiencies . Inactivation of the K-region epoxide (in terms of its mutagenicity in the presence of glutathione) required extremely little enzyme, about 1000 times less than for the diol-epoxide . On a molar basis, glutathione transferase X (followed by C greater than A greater than or equal to B) was clearly the most efficient enzyme in inactivating both substrates and also more efficient than were three other purified enzymes (microsomal epoxide hydrolase, cytosolic epoxide hydrolase, and dihydrodiol dehydrogenase) previously investigated in this test system . Taking into account the amounts of enzyme present in rat liver, the glutathione transferases C and X were most effective in inactivating the epoxides examined . Thus, the newly discovered glutathione transferase X appears to be of substantial significance in the inactivation of two structural prototypes of epoxides derived from polycyclic aromatic hydrocarbons, a K-region epoxide and a non-bay-region vicinal diol-epoxide. Gene, 1983 Dec, 26(2-3), 147 - 58 Use of M13mp phages to study gene regulation, structure and function: cloning and recombinational analysis of genes of the Salmonella typhimurium histidine operon; Artz S et al.; A restriction map was determined for a phi 80 lambda dhis transducing phage DNA carrying the Salmonella typhimurium histidine operon . DNA fragments containing the promoter/regulatory region and the first two structural genes of the histidine operon (hisOGD) were identified by their ability to direct regulated synthesis of histidinol dehydrogenase (product of hisD) in a coupled in vitro protein synthesizing system . A 3.1-kb SalI-EcoRI restriction fragment containing the hisOGD region, was subcloned into phage M13mp8 and M13mp9 RF DNAs . Methods are described for shuttling mutant and wild-type bacterial DNA sequences between the M13mp::his phage and host bacterial genomes . Of novel importance is the use of the phage M13 gene II amber mutation to obtain integration of the M13mp::his phage genome into the homologous his region of the bacterial chromosome following transduction of recipients lacking an amber suppressor . This method can be used to facilitate allele replacement with genes carried on M13 transducing phages. Cancer Lett, 1983 Dec, 21(2), 211 - 7 Microsomal P-450 induction by some secondary products from thermal oxidation of dietary lipids: epidermal hyperplasia, mutagenicity and cytochrome P-450 activities; Crawford L et al.; Distillable secondary products from roasted fowl were found to be cytotoxic but not mutagenic when assayed with Salmonella typhimurium strains TA98, TA100 and TA1537 . A crudely separated fraction of the volatiles produced focal hyperplasia and damage to the epidermis of the backs of mice . The volatiles also caused an apparent synthesis of non-constitutive forms of rat hepatic cytochromes P-450 which metabolize benzo{a}pyrene B {a}P differently from the constitutive P-450. J Immunol, 1983 Dec, 131(6), 3014 - 20 Genetic control of the innate resistance of mice to Salmonella typhimurium: Ity gene is expressed in vivo by 24 hours after infection; Swanson RN et al.; The early response of inbred mice to infection with S . typhimurium is controlled by the mouse Chromosome 1 locus, Ity . To better understand the expression of this gene, the initial interactions between the reticuloendothelial system (RES) and i.v . injected salmonellae were compared in resistant (Ityr) and susceptible (Itys) mice . In both mouse strains 99% of the bacteria was cleared from the blood within 2 hr, and uptake of S . typhimurium by splenic and hepatic macrophages was similar regardless of Ity genotype . In vivo phagocytosis of bacteria was followed by a 30 to 60% decline in viable bacteria, which was attributed to the bactericidal activity of RES macrophages . Experiments with radiolabeled S . typhimurium strains TML and TML/TS27 (a temperature-sensitive mutant) confirmed that the efficiency of this early phase killing was not under Ity control . Despite the equivalent uptake and initial bactericidal activity by resident macrophages, bacterial numbers in the RES organs of Itys mice were significantly greater than in Ityr mice by approximately 24 hr after infection . These data suggest that Ity regulates the level of surviving intracellular bacteria that accumulate within resident macrophages of the liver and spleen. J Bacteriol, 1983 Dec, 156(3), 1249 - 62 Genetic analysis of the proBA genes of Salmonella typhimurium: physical and genetic analyses of the cloned proB+ A+ genes of Escherichia coli and of a mutant allele that confers proline overproduction and enhanced osmotolerance; Mahan MJ et al.; Because of the fact that proline overproduction relieves the inhibitory effects of high external osmotic strength in a number of procaryotes, we wished to clone a mutant allele, pro-74, that confers proline overproduction and enhanced osmotolerance on Salmonella typhimurium and Escherichia coli . Therefore, the pro-74 allele, originally located on an E . coli episome F'128, was cloned into pBR322 . In a parallel experiment, the wild type proB+ A+ genes of E . coli were also cloned from F'128 into pBR322 . Both the pro-74 and the proB+ A+ alleles were obtained on a 10.4-kilobase-pair fragment that also contained the unrelated phoE gene . Strains carrying either the wild-type proB+ A+ or the pro-74 alleles on pBR322 grew more slowly, both in minimal medium and media of elevated osmotic strength, than strains carrying the same alleles on the low-copy plasmid, F'128, indicating that some gene in the cloned region is deleterious in high copy . We constructed Tn5 insertion mutations in the proB and the proA genes of E . coli, carried on F'128 in S . typhimurium . Using P22 transduction in S . typhimurium, we transferred these proB and proA::Tn5 insertions from F'128 into the cloned proBA genes on pBR322 . From the restriction maps of the plasmids thus generated, we determined the approximate locations of the proB and the proA genes . We also performed complementation tests of S . typhimurium and E . coli proB and proA mutants by using the F'128 proB and proA::Tn5 insertions . These tests revealed that the proBA genes of S . typhimurium form an operon, whose direction of transcription is from proB to proA . They also indicated that in S . typhimurium, as in E . coli, the proB+ gene encodes gamma-glutamyl kinase, and the proA+ gene encodes gamma-glutamyl phosphate reductase . Complementation tests also indicated that the pro-74 mutation is either in the proB structural gene, or its promoter-operator. Genetics, 1983 Dec, 105(4), 801 - 11 Genetic mapping of IS200 copies in Salmonella typhimurim strain LT2; Lam S et al.; The wild-type Salmonella typhimurium strain LT2 contains six copies of the insertion sequence element IS200 which is unique to Salmonella . We have determined the chromosomal locations of all six copies of IS200 in strain LT2 . This was done by mapping the positions of Tn10 elements inserted near each copy of IS200 . Such Tn10 insertions were detected by Southern hybridization as IS200-containing restriction fragments with altered electrophoretic mobility . The copies are located at quite evenly spaced sites in the chromosome . Some are found in regions with many known genes; others are in regions with few known functions . There is no indication of a possible function for IS200 . The method described here should be applicable to the mapping of IS elements in general. Mutat Res, 1983 Dec, 124(3-4), 191 - 200 Mutagenicity of diesel-exhaust particle extract, 1-nitropyrene, and 2,7-dinitrofluorenone in Salmonella typhimurium under various metabolic activation conditions; Kohan M et al.; The mutagenic activities of 1-nitropyrene (1-NP), 2,7-dinitrofluorenone (2,7-DNF), and a diesel-exhaust extract were compared using the Salmonella typhimurium plate-incorporation assay . Each sample was tested with and without a 9000 X g liver homogenate (S9), both with and without an NADPH-generating system . The samples were also treated with the microsome fraction of S9, cytosol fraction of S9, boiled S9, bovine serum albumin (BSA), and boiled BSA . Salmonella tester strains TA98 and TA98FR1 were used in all treatments; TA98/1,8DNP6 was used to test mutagenic activity without activation . Without the NADPH-generating system, the samples generally had less mutagenic activity than samples treated with the NADPH-generating system . The addition of the NADPH-generating system resulted in marked increases in mutagenic activity of 1-NP in the microsome and S9 treatments, and of all 3 samples in the cytosol fraction treatment . These results indicate that although protein binding reduced the mutagenic activity of diesel-exhaust extract and 1-NP, microsomal activation increased the mutagenic activity of 1-NP . Because 1-NP and 2,7-DNF contributed less than 1.5% of the mutagenic activity of the diesel-exhaust extract, the response to diesel exhaust was not typified by these compounds. J Bacteriol, 1983 Dec, 156(3), 1344 - 8 recA-dependent recombination between rRNA operons generates type II F' plasmids; Blazey DL et al.; The formation of type II F' ilv cya metE plasmids from the Salmonella typhimurium Hfr strain SA722 occurs by general recombination between repeated rrn. Can J Microbiol, 1983 Dec, 29(12), 1731 - 5 Attachment of Salmonella to mammalian cells in vitro; Mintz CS et al.; The attachment of Salmonella typhimurium strain PHL67342 to several mammalian tissue culture cell lines was investigated . Strain PHL67342 failed to attach in significant numbers to the Buffalo green monkey (BGM), swine testicular (ST), and HeLa cell lines . Significant attachment was observed with the Henle intestinal cell line . Log-phase cells of strain PHL67342 attached in greatest numbers to the Henle cells after 45 min of incubation at 37 degrees C . Attachment to the Henle cells was not affected by D-mannose or D-galactose, but was markedly inhibited by high concentrations of alpha-methyl-D-mannoside . Also, Salmonella lipopolysaccharide had no effect on the attachment of strain PHL67342 to the Henle cells . Fimbriae were not detected on the bacterial cells used in the adherence experiments . These results suggest that some bacterial factor(s) other than fimbriae and lipopolysaccharide mediate the attachment of strain PHL67342 to the Henle cells. Mol Biol Evol, 1983 Dec, 1(1), 57 - 66 Evolution of antibiotic resistance genes: the DNA sequence of a kanamycin resistance gene from Staphylococcus aureus; Gray GS et al.; The kanamycin resistance gene from Staphylococcus aureus has been sequenced and its structure compared with similar genes isolated from Streptomyces fradiae and from two transposons, Tn5 and Tn903, originally isolated from Klebsiella pneumoniae and Salmonella typhimurium, respectively . The genes are all homologous but, since their common ancestor, have undergone extensive divergence, with more than 43% divergence between the closest pair . The phylogeny of the genes cannot be made congruent to the phylogeny of the taxa from which they were isolated without requiring rather improbable differences in rates . One is therefore led to conclude that there have been multiple occurrences of gene transfer between these species . Thus, although they are homologous, they are neither orthologous nor paralogous . It is suggested that homologous genes of this type be called xenologous. J Biol Chem, 1983 Nov 25, 258(22), 13665 - 72 Rates of ligand binding to periplasmic proteins involved in bacterial transport and chemotaxis; Miller DM 3rd et al.; The ligand reactions of three binding proteins involved in bacterial transport and chemotaxis have been examined by stopped flow, rapid mixing techniques . The processes measured were: L-arabinose, D-galactose, and D-fucose binding to the Escherichia coli L-arabinose-binding protein; L-histidine binding to the Salmonella typhimurium L-histidine-binding protein; and D-maltose, maltotriose, cyclic maltohexaose, and cyclic maltoheptaose binding to the E . coli D-maltose-binding protein . Changes in tryptophan fluorescence were monitored, and the resultant time courses were analyzed quantitatively in terms of a simple one-step binding process . The fitted association rate constants for sugar binding are all about 1-3 X 10(7) M-1 s-1; variation in the affinity constants is expressed primarily by changes in the dissociation rate constants, 1-100 s-1 . The sugar-binding proteins react at equal rates with the alpha and beta anomeric forms of their substrates . The ligand dissociation rates measured in vitro are consistent with the corresponding Vmax values observed for in vivo active transport . The association rate constant for the L-histidine-binding protein is 5-10 times greater than the corresponding rate constants for the sugar-binding proteins . A similar, large bimolecular rate, approximately 1 X 10(8) M-1 s-1, has been observed for the E . coli L-glutamine-binding protein (Weiner, J . H., and Heppel, L . A . (1971) J . Biol . Chem . 246, 6933-6941) and appears to reflect favorable electrostatic interactions between the charged amino acid and the surface of the protein molecule. Biochim Biophys Acta, 1983 Nov 23, 735(3), 337 - 40 Transport of 2-methyl-4-amino-5-hydroxymethylpyrimidine by Salmonella typhimurium; Bellion E et al.; The transport of 2-methyl-4-amino-5-hydroxymethylpyrimidine (MAHMP) by Salmonella typhimurium was studied using synthetic {methyl-3H3}MAHMP . It was found that an active transport system existed for MAHMP, having Km of 0.07 microM and Vmax 45 nmol.min-1.(g dry wt . cells)-1, that required glucose as a source of energy and was pH and temperature dependent . Uptake was inhibited by cyanide, azide, N-ethylmaleimide, 2,4-dinitrophenol and carbonyl cyanide m-chlorophenylhydrazone . Uptake was also weakly inhibited by oxythiamine, but not by thiamine, 2-methyl-4-amino-5-aminomethylpyrimidine, or 4-amino-5-hydroxymethylpyrimidine, indicating that the transport system is specific for MAHMP. Biochim Biophys Acta, 1983 Nov 23, 735(3), 331 - 6 Transport of thiamine and 4-methyl-5-hydroxyethylthiazole by Salmonella typhimurium; Bellion E et al.; The transport of thiamine and 4-methyl-5-hydroxyethylthiazole (MHET), its thiazole moiety, was studied using whole cells of Salmonella typhimurium . It was found that the bacteria possessed an active transport system for thiamine that had Km 0.21 microM and Vmax 33 nmol.min-1.(mg dry wt . cells)-1 . Transport of thiamine was glucose dependent, whereas MHET uptake was dependent on both glucose and 2-methyl-4-amino-5-hydroxymethylpyrimidine (MAHMP), the pyrimidine moiety of thiamine . Uptake of both thiamine and MHET was severely curtailed by cyanide, azide, N-ethylmaleimide and carbonyl cyanide m-chlorophenylhydrazone . Oxythiamine inhibited thiamine, but not MHET, uptake and thiamine slightly inhibited MHET uptake . 2-Methyl-4-amino-5-methoxymethylpyrimidine and 4-amino-5-hydroxymethylpyrimidine were unable to replace MAHMP as stimulators of MHET uptake, but 2-methyl-4-amino-5-aminomethylpyrimidine was marginally effective in this regard . Similar results were obtained with attempts to replace MAHMP as a growth requirement for a purD mutant of Salmonella typhimurium . MHET uptake showed saturation kinetics only in the presence of MAHMP, and is not otherwise actively transported. Biochim Biophys Acta, 1983 Nov 17, 741(2), 180 - 96 Sequence specificity of tRNA-modifying enzymes . An analysis of 258 tRNA sequences; Tsang TH et al.; The specificity and recognition of tRNA-modifying enzymes may be accounted for in part by nucleotide sequences which are localized next to the modifiable nucleoside . In order to determine the sequence specificity of tRNA-modifying enzymes, we have surveyed 55 published tRNA sequences from Escherichia coli, Salmonella typhimurium and T4 phage . For each modified nucleoside, the nucleotide sequence surrounding the modification site was determined for all tRNAs known to contain the modified nucleoside . Subsequently all tRNAs not containing the modified nucleoside were examined for the absence of the putative recognition site . We present the detailed analysis of 12 modified nucleosides for which we found a strong correlation between the modified nucleoside and the local nucleotide sequence . This suggests that these sequences may be recognition sites for tRNA-modifying enzymes . For each of the 12 modified nucleosides we have identified a recognition sequence present in the tRNA set containing the modification and not in the set without it . All 203 other published tRNA sequences were then examined to see if the sequence specificity rules apply to other organisms, including both prokaryotes and eukaryotes . In several cases a good adherence was found, indicating conservation of the putative recognition sequences. Klin Wochenschr, 1983 Nov 15, 61(22), 1153 - 7 {Myocarditis caused by Salmonella typhimurium}; Gotz M et al.; A 53-year-old man died on the eight day of an acute enteritis caused by Salmonella typhimurium . Clinical signs of shock were pronounced; the electrocardiogram, initially not pathological, showed a peripheral low voltage with decreased R-amplitudes and distinct disturbances in repolarization . Laboratory findings demonstrating Salmonella typhimurium in blood cultures and stools and 45% band forms in the differential count were remarkable . The autopsy showed deep, fibrin-covered ulcera in the colon area and a dense submucous lympho-histiocyte cell infiltration . Histologically, a granulomatous myocarditis of the left ventricle with lymphocyte and histiocyte infiltration and a focal myolysis was observed . In the right ventricle, however, only a minimal interstitial edema was found. J Biol Chem, 1983 Nov 10, 258(21), 12947 - 51 Position of ester groups in the lipid A backbone of lipopolysaccharides obtained from Salmonella typhimurium; Qureshi N et al.; Lipopolysaccharides extracted from the heptoseless mutant of Salmonella typhimurium G30/C21 were hydrolyzed with either 0.1 N HCl at 100 degrees C or treated twice with 20 mM sodium acetate, pH 4.5, at 100 degrees C for 45 min and finally purified by preparative thin layer chromatography to yield a structural series of mono- and diphosphoryl lipid A, respectively . Positive ion fast atom bombardment mass spectrometry of the diphosphoryl lipid A TLC-3 (a highly acylated major band) showed a major component with (M + H)+ ion of mass 1798, which fragmented to yield a (M - H2PO4)+ ion of mass 1700 . Cleavage at the glycosidic bond gave rise to an oxonium ion fragment of mass 1087 . In conjunction with other studies, this establishes the molecular formula and Mr of the major component to be C94H178N2O25P2 and 1797.2 (as the free acid), respectively . Similar analysis of monophosphoryl lipid A TLC-3 produced an (M + H)+ peak at m/z 1718, (M + Na)+ adduct peak at m/z 1740, and a fragment of mass 1087 . The spectrum of monophosphoryl lipid A TLC-5 was devoid of the m/z 1087 peak and instead contained the phosphorylated oxonium ion of mass 876 . This fragment ion is assigned as the distal subunit, and these results show that the distal subunit of the major lipid A TLC-3 contains two hydroxymyristoyl, one myristoyl, and one lauroyl residues, whereas the reducing end subunit contains two hydroxymyristoyl groups . A revised structure of the lipid A backbone in lipopolysaccharides of S . typhimurium is proposed. J Biol Chem, 1983 Nov 10, 258(21), 12801 - 3 Complete structure of lipid A obtained from the lipopolysaccharides of the heptoseless mutant of Salmonella typhimurium; Takayama K et al.; A highly purified monophosphoryl lipid A, TLC-3 fraction obtained from the lipopolysaccharides of the heptoseless mutant Salmonella typhimurium G30/C21 was converted to the dimethyl pentatrimethylsilyl derivative and analyzed by proton NMR spectroscopy at 400 MHz . Substantial downfield shifts of the resonances for protons at the 3- and 3'-carbons of the glucosamine disaccharide to 5.06 and 5.15 ppm, respectively, occurred from the normal range of 3.5-4.1 ppm, indicating that these two positions on the sugar rings were acylated . Significant downfield shift of the resonances for protons at the 4- and 6'-carbons did not occur, indicating the absence of acyl groups at these two positions . Since positive ion fast atom bombardment mass spectrometry previously established the presence of hydroxymyristoyl and myristoxymyristoyl esters at the reducing end and distal subunits, respectively, these acyl groups must be attached to the oxygen of the corresponding 3- and 3'-carbons of lipid A . With these results, we can now describe the complete structure of the monophosphoryl lipid A, TLC-3 from S . typhimurium. J Biol Chem, 1983 Nov 10, 258(21), 12982 - 7 Evidence for a small catalytic domain in the adenylate cyclase from Salmonella typhimurium; Leib TK et al.; Deletions of large portions of the carboxyl-terminal end of the adenylate cyclase (ATP pyrophosphate lyase (cyclizing), EC 4.6.1.1) from Salmonella typhimurium do not significantly affect the enzymatic activity exhibited by the shortened polypeptide . The deletion mutations were generated by nuclease Bal31 digestion from the 3'-end of the cya gene fragment cloned by Wang et al . (Wang, J . Y.-J., Clegg, D . O., and Koshland, D.E . (1981) Proc . Natl . Acad . Sci . U.S.A . 78, 4684-4688); the shortened cya genes were inserted in pBR322 and used to transform a cya- strain of Escherichia coli . The original gene fragment encodes for an enzymatically active polypeptide having an apparent molecular weight of 77,000 . Mutant polypeptides as small as 46,000 Da were found to retain significant enzymatic activity and to confer several cya+ phenotypes on the E . coli host . More extensive deletions resulting in polypeptides as small as 33,000 Da did not have assayable amounts of adenylate cyclase activity, but the biochemical properties of the transformed cya- host implicate the presence of low levels of enzymatic activity . These data suggest that the structure of the intact enzyme is composed of discrete functional domains . Such a structure for this adenylate cyclase should both facilitate investigations of the chemical mechanism of the reaction and allow structure-function relationships in this physiologically important enzyme to be investigated on a molecular level. Clin Exp Immunol, 1983 Nov, 54(2), 319 - 26 The role of the spleen in the protective effect of C-reactive protein in Streptococcus pneumoniae infection; Nakayama S et al.; C-reactive protein (CRP) is an acute phase serum protein in man which activates complement and has opsonic activity . We have reported that prior injection of CRP into mice can increase their survival following intravenous challenge with Streptococcus pneumoniae type 3 or 4 . In this study the conditions required for protection, and the role of hepatic and splenic clearance of bacteria have been examined . Protection against lethal infection was observed with a minimum dose of 25-50 micrograms CRP per mouse . CRP was most effective when administered between 6 h before and 2 h after challenge . CRP treated mice were not protected against infection with Salmonella typhimurium, LT-2, an organism which does not bind CRP . Mice depleted of C3 by treatment with cobra venom factor were protected against S . pneumoniae infection by CRP . Pre-treatment of mice with CRP did not increase the rate of clearance of viable S . pneumoniae from the bloodstream but did increase splenic and decrease hepatic clearance of radiolabelled bacteria in both normal and complement depleted mice . Although these findings suggest a role for the spleen in CRP protection, mice which had been splenectomized were also protected against lethal pneumococcal infection by CRP treatment. Mutat Res, 1983 Nov, 124(2), 129 - 43 Genotoxicity studies with a blend of zinc dialkyldithiophosphate lubricant additives; Brooks TM et al.; The mutagenic activity of a blend of primary zinc dialkyldithiophosphate lubricant additives suspended in process oils and a blend of the process oils alone was investigated in agar layer cultures of Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100, both with and without the incorporation of a rat liver microsomal activation system (S9) . Zinc dimethyldithiocarbamate was tested as a structurally-related model bacterial mutagen, and, in additional control experiments, the mutagenic activity of zinc dimethyldithiocarbamate and benzo{a}pyrene was investigated in combination with the blend of process oils in selected bacterial tester strains . Transformation frequencies of BHK cells were determined by colony growth in soft agar culture following treatment with a blend of the zinc dialkyldithiophosphate lubricant additives suspended in process oils, a blend of the process oils, 7,12-dimethylbenzanthracene, both in the presence and in the absence of a blend of the process oils, or zinc dimethyldithiocarbamate . All experiments incorporated a rat liver microsomal activation system (S9) . Application of a blend of the zinc dialkyldithiophosphate additives or a blend of the process oils used in their manufacture did not increase the reverse mutation frequencies of Salmonella typhimurium TA1535, TA1537, TA1538, TA98 or TA100, or significantly increase the transformation frequency of BHK cells under the experimental conditions described . Zinc dimethyldithiocarbamate increased the reverse mutation frequency in some bacterial tester strains, but did not significantly increase the transformation frequency of BHK cells under the described experimental conditions . The addition of the blend of the process oils in combination with the control materials, zinc dimethyldithiocarbamate or benzo{a}pyrene had an inhibitory effect on the mutagenic activity at high doses of each in the bacterial assays, and in the BHK assay the transforming ability of 7,12-dimethylbenzanthracene was suppressed in the presence of the blend of the process oils . Thus, the additive materials showed no evidence of genotoxic activity in the bacterial mutation assays, or in the BHK transformation assay under the experimental conditions described. Mutat Res, 1983 Nov, 124(2), 121 - 8 Mutagenicity studies of ambient airborne particles . II . Comparison of extraction methods; Krishna G et al.; Organic materials were extracted with acetone from airborne particles by shaking, soxhletion and sonication for varying durations . 4-h, 1-h and 1/8-min extractions by shaking, soxhletion and sonication, respectively gave maximum his+ revertants with the Ames Salmonella/microsome assay . In a comparative study of extraction methods, sonication gave the highest and soxhletion the lowest mutagenic response . It appears that sonication with acetone is the best procedure for the extraction of mutagens from airborne particles as shown by Ames assay and Arar assay systems in Salmonella typhimurium. Cancer Lett, 1983 Nov, 21(1), 63 - 8 Mutagenic activity of N-nitrosomethamphetamine and N-nitrosoephedrine; Shimizu H et al.; The mutagenicity of N-nitrosomethamphetamine (NMA) and N-nitrosoephedrine (NEP), which were synthesized in our laboratory, was examined by the modified pre-incubation method of Ames assay using Salmonella typhimurium TA100 and TA98 . Both nitroso compounds showed significant mutagenic activity in the presence of hamster S9. Chem Biol Interact, 1983 Nov, 47(2), 175 - 94 Interrelationships in mice of antipyrine half-life, hepatic monooxygenase activities and liver S9-mediated mutagenicity of aflatoxin B1, benzo{alpha}pyrene 7,8-dihydrodiol, 2-acetylaminofluorene and N-nitrosomorpholine; Roberfroid MB et al.; To evaluate the predictive value of serum antipyrine half-life AP(T1/2) as an index of hepatic carcinogen metabolism, groups of C57BL/6 and DBA/2 mice were treated with various inducers and inhibitors of cytochrome P-450-dependent monooxygenases (pregnenolone-16 alpha-carbonitrile (PCN), phenobarbital (PB), 5,6-benzoflavone (5,6-BF), 3-methylcholanthrene (MC), disulfiram (DIS), 7,8-BF) . Groups of mice were also given ethanol (3% in drinking water) for 12 days . Within each group, mean serum AP-(T1/2) was compared with (i) the in vitro activity of hepatic microsomal benzo{alpha}pyrene (BP) 3-hydroxylase, 2-acetylaminofluorene (AAF)-N-hydroxylase and aldrin monooxygenase, and (ii) the liver S9-mediated mutagenicity of aflatoxin B1 (AFB), trans-7,8-dihydro-7,8-dihydroxybenzo{alpha}pyrene (BP 7,8-diol), 2-acetylaminofluorene and N-nitrosomorpholine (NMOR) in Salmonella typhimurium strains . Serum AP(T1/2) was only correlated negatively with the activity of BP 3-hydroxylase (P less than 0.001) and aldrin monooxygenase (P less than 0.001) . No statistically significant correlation was found between serum AP(T1/2) and liver S9-mediated mutagenicity for any of the four carcinogens . On the basis of these results, we conclude that serum AP(T1/2) may not be a reliable index of the capacity of liver to convert carcinogens into reactive intermediates. Can J Microbiol, 1983 Nov, 29(11), 1583 - 8 The epidemic spread of Salmonella typhimurium phage type 10 in Canada (1970-1979); Khakhria R et al.; The frequency of Salmonella typhimurium phage type 10 across Canada was monitored during the period 1970-1979 . Phage type 10 isolations increased from 1.2% in 1970 to 68.8% in 1979 among isolates from human sources and from 1.5 to 30.6% in isolates from nonhuman sources . Examination of food-poisoning outbreaks and a study of the animal-host associations of phage type 10 revealed that contaminated poultry products appear to be the most common sources of human infections . The majority (89.3%) of S . typhimurium phage type 10 strains were sensitive to antibiotics . Of the resistant strains, 73.3% were resistant to single antibiotics and 26.7% were multiresistant . Thirty-three different patterns of antibiotic resistance were observed . A number of the resistance determinants were transferable by conjugation and the R plasmids were found to belong to the incompatibility groups HI1, FII, N, I alpha, and C. J Gen Microbiol, 1983 Nov, 129 ( Pt 11), 3395 - 400 Generalized transduction between Salmonella typhi and Salmonella typhimurium by phage j2 and characterization of the j2 plasmid in Escherichia coli; Mise K et al.; Phage j2, a P1-like phage in Salmonella typhi, was heteroimmune to phage P1 and existed in the lysogenic state as a plasmid of molecular size 58.6 MDal . The phage j2 plasmid was incompatible with the P1 plasmid (IncY group) . A j2-sensitive mutant of Salmonella typhimurium LT2 was isolated by transduction of j2Ap phage into LT2 followed by curing of the prophage . The mutant was used