J Bacteriol, 1984 Oct, 160(1), 122 - 30 Genetic characterization and molecular cloning of the tripeptide permease (tpp) genes of Salmonella typhimurium; Gibson MM et al.; Of the three bacterial peptide transport systems only one, the oligopeptide permease, has been characterized in any detail . We have now isolated Salmonella typhimurium mutants deficient in a second transport system, the tripeptide permease (Tpp), using the toxic peptide alafosfalin . Alafosfalin resistance mutations map at three loci, the gene encoding peptidase A (pepA) and two transport-defective loci, tppA and tppB . Locus tppA has been mapped to 74 min on the S . typhimurium chromosome, cotransducible with aroB, and is a positive regulator of tppB . Locus tppB maps at 27 min in the cotransduction gap between purB and pyrF . We cloned tppB, the structural locus for the tripeptide permease . Two simple methods are described for mapping the location of cloned DNA fragments on the chromosome of S . typhimurium. Infect Immun, 1984 Oct, 46(1), 231 - 6 Brucellacidal activity of human and bovine polymorphonuclear leukocyte granule extracts against smooth and rough strains of Brucella abortus; Riley LK et al.; The microbicidal activities of freeze-thaw and high-salt extracts of human and bovine polymorphonuclear leukocyte (PMN) granules were tested against a smooth intermediate strain (45/0) and a rough strain (45/20) of Brucella abortus which differ in virulence and survival within PMNs . Freeze-thaw extracts of human PMN granules were more brucellacidal than high-salt extracts when supplemented with hydrogen peroxide (H2O2) and potassium iodide (KI), whereas the opposite was found with freeze-thaw and high-salt extracts of bovine PMN granules . There was no oxygen-independent killing of either the smooth or rough strain of B . abortus by amounts of granule extracts which caused 100% killing of a deep rough mutant (Re) of Salmonella typhimurium . The oxygen-dependent brucellacidal activity of granule extracts was dependent on concentrations of myeloperoxidase (MPO) units, H2O2, and KI . Maximal brucellacidal activity was observed at pH 5.5 to 6.0 . The smooth strain, 45/0, was more resistant to oxygen-dependent killing by granule extracts than was the rough strain, 45/20 . Granule extracts were more brucellacidal than purified MPO at equivalent levels of MPO enzyme units, suggesting that at least one other reaction enhances killing by the MPO-H2O2-I- system. Biochemistry, 1984 Sep 25, 23(20), 4767 - 73 Phosphorothioate analogues of 5-phosphoribosyl 1-diphosphate: synthesis, purification, and partial characterization; Smithers GW et al.; The 1-phosphorothioate analogues of 5-phosphoribosyl 1-diphosphate (P-Rib-PP) have been prepared enzymatically, in reactions catalyzed by P-Rib-PP synthetase from Salmonella typhimurium . 5-Phosphoribosyl 1-O-(2-thiodiphosphate) (P-Rib-PP beta S) was synthesized from ribose 5-phosphate (Rib-5-P) and the Mg2+ complex of adenosine 5'-O-(3-thiotriphosphate) . The SP and RP diastereomers of 5-phosphoribosyl 1-O-(1-thiodiphosphate) (P-Rib-PP alpha S) were synthesized from Rib-5-P and the Mg2+ complex of adenosine 5'-O-(2-thiotriphosphate) (ATP beta S) (SP diastereomer, delta-configuration) and the Cd2+ complex of ATP beta S (RP diastereomer, delta-configuration), respectively . The strategy for the synthesis and stereochemical assignment of the P-Rib-PP alpha S diastereomers was based on the specificity of P-Rib-PP synthetase for the (delta)-beta, gamma-bidentate metal-nucleotide substrate and the stereochemical course of the synthetase reaction, leading to inversion of configuration at the P beta atom of the nucleotide {Li, T . M., Mildvan, A . S., & Switzer, R . L . (1978) J . Biol . Chem . 253, 3918-3923}, and the known configurations of the Mg2+ and Cd2+ beta, gamma-bidentate complexes of the ATP beta S diastereomers {Jaffe, E . K., & Cohn, M . (1979) J . Biol . Chem . 254, 10839-10845} . The P-Rib-PP analogues were purified by gradient elution from DEAE-Sephadex and characterized by chemical analysis and 31P nuclear magnetic resonance {Smithers, G . W., & O'Sullivan, W . J . (1984) Biochemistry (following paper in this issue)} . A preliminary account of their interaction with human brain hypoxanthine phosphoribosyltransferase and yeast orotate phosphoribosyltransferase (OPRTase) is described.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1984 Sep 10, 259(17), 10983 - 8 Oxygen taxis and proton motive force in Salmonella typhimurium; Shioi J et al.; The aerotactic response of Salmonella typhimurium SL3730 has been quantitatively correlated with a change in the proton motive force (delta p) as measured by a flow-dialysis technique . At pH 7.5, the membrane potential (delta psi) in S . typhimurium changed from -162 +/- 13 to -111 +/- 15 mV when cells grown aerobically were made anaerobic, and it returned to the original value when the cells were returned to aerobiosis . The delta pH across the membrane was zero . At pH 5.5, delta psi was -70 mV in aerobiosis and -20 mV in anaerobiosis, and delta pH was -118 and -56 mV for aerobic and anaerobic cells, respectively . A decrease in delta p resulted in increased tumbling, and an increase in delta p resulted in a smooth swimming response at either pH . Inhibition of aerotaxis at pH 7.5 by various concentrations of KCN correlated with a decreased delta p, due to a decreased delta psi in aerobiosis and little change in delta psi in anaerobiosis . At concentrations up to 100 mM, 2,4-dinitrophenol decreased delta psi, but did not inhibit aerotaxis because the difference between delta psi in aerobic and anaerobic cells remained constant . Considered as a whole, the results indicate that aerotaxis in S . typhimurium is mediated by delta p. Carcinogenesis, 1984 Sep, 5(9), 1179 - 81 Mutagenicity of smoke condensate of bidi--an indigenous cigarette of India; Shirname LP et al.; The possible mutagenicity of bidi smoke condensate (BSC), an indigenous form of an Indian cigarette has been studied using three short-term test systems . It was seen that BSC caused frame-shift mutations in Salmonella typhimurium strains TA 98 and TA 1538 and required metabolic activation . In the mammalian test systems, BSC induced 8 azaguanine resistant mutations in V79 Chinese hamster cells in the presence of S9 mixture and induced elevated frequencies of micronucleated erythrocytes in the bone marrow of Swiss mice. Cancer Res, 1984 Sep, 44(9), 3736 - 43 Pharmacological and preclinical toxicological studies of 1,2-diaminocyclohexane(isocitrato)platinum(II); Macquet JP et al.; The antitumor activity of a new highly water-soluble platinum derivative, (1,2-diaminocyclohexane)(isocitrato)platinum(II) (NSC 350602; PHIC), was studied in L1210 leukemia cells inoculated into mice . PHIC was found to be active for i.p . graft-i.p . treatment, i.p . graft-i.v . treatment, and i.v . graft-p.o . treatment . A significant activity was observed on early and advanced L1210 leukemia even when the treatment was delayed 6 days after the graft . A comparison between the activities of PHIC, cisplatin (NSC 119875), and (4-carboxyphthalato)(1,2-diaminocyclohexane)-platinum(II) (NSC 271674; DACCP) for i.p . graft-i.p . treatment indicated that the highest activity was observed for divided doses rather than single dose in the case of PHIC and DACCP and not for cisplatin . Under these conditions, PHIC gave larger treated versus control survival time values or a greater number of surviving animals than did cisplatin and DACCP . No cross-resistance between PHIC and cisplatin could be detected in L1210 leukemia cells resistant to cisplatin . Mutagenicity studies on Salmonella typhimurium revealed that PHIC is far less mutagenic than cisplatin on TA100 and TA98 strains . Other pharmacological parameters, such as growth inhibition rate of cultured L1210 cells, penetration, and DNA binding in L1210 cells inoculated in mice, were compared for PHIC and cisplatin together with their in vitro rates of hydrolysis and platinum:DNA adducts . No nephrotoxicity was detected with PHIC at the maximum nonlethal dose level in mice in contrast to results with cisplatin . A preclinical study was conducted in baboons at 100, 150, and 200 mg/kg . No nephrotoxicity could be detected at a dose of 100 mg/kg without prehydration for six courses at 3-week intervals . At 200 mg/kg, an increase of blood creatinine was controlled by prehydration . Gastrointestinal toxicity was mild during the three regimens . Phase I clinical trials are under way. Sci Total Environ, 1984 Sep, 38, 275 - 81 Study of mutagenic pollution in Bombay; Shenoy CN et al.; Airborne suspended particulate matter (SPM) from seven areas in and around Bombay city were collected over glass fibre filters (0.8 micrometer porosity) . The chemicals from the SPM were extracted in dimethylsulfoxide and distilled water and were further tested for mutagenicity by Ames' test using for five mutants of Salmonella typhimurium . Of the seven areas studied, only four exhibited mutagenicity, which was confirmed by dose-response assays using the mutant strain TA 100 . The very high mutagenicity observed in central Bombay correlates with the higher incidence of respiratory tract diseases in the resident population. J Med Chem, 1984 Sep, 27(9), 1161 - 6 Potential antitumor agents: synthesis and biological properties of aliphatic amino acid 9-hydroxyellipticinium derivatives; Auclair C et al.; Aliphatic amino acids glycine, alanine, valine, and leucine were conjugated to the antitumor drug N2-methyl-9-hydroxyellipticinium (NMHE) through a peroxidase-catalyzed oxidation reaction . NMR studies of the adducts so obtained have indicated (i) that the amino acids were linked to NMHE between the nitrogen of their primary amine and the C-10 position of the ellipticine ring and (ii) that a double bond was present between the nitrogen and the alpha-carbon of the amino acid moiety . All amino acid-NMHE adducts exhibit a higher lipophilic property than the parent compound (NMHE) directly correlated with the length of the aliphatic chain of the amino acids . The adducts interact with DNA through an intercalating process with apparent binding constant ranging from 2 X 10(5) to 5 X 10(5) M-1 at pH 7.40 . The presence of the amino acid moiety linked to NMHE results (i) in a slight decrease of the cytotoxicity on L1210 cells in vitro (ID50 ranged from 0.20 to 0.50 microM) as compared to NMHE (ID50 = 0.05 microM), (ii) in a decrease of the antitumor efficiency in vivo against L1210 leukemia for leucine-NMHE and valine-NMHE (ILS at LD0/2 = 35% and 31%, respectively), (iii) in a suppression of the antitumor activity for alanine-NMHE and glycine-NMHE (ILS less than 25%), (iv) in a strong increase in the bacteriostatic activity on the quaternary ammonium sensitive Escherichia coli BL101 strain and on Salmonella typhimurium TA98 strain . The bacteriostatic effect is directly correlated with the lipophilic property of the drugs . These findings are discussed in terms of a structure-activity relationship. Zh Mikrobiol Epidemiol Immunobiol, 1984 Sep, (9), 43 - 9 {Isolation of Salmonella typhimurium enterotoxin in partially purified form and study of its properties}; Fluer FS; S . typhimurium enterotoxin, partially purified in accordance with our scheme (salting out with 75% ammonium sulfate, dialysis and gel filtration in a column with Sephadex G-150, followed by electrofocusing), showed enterotoxic activity in the intestinal loop of a rabbit and yielded the positive result in the cutaneous test . S . typhimurium enterotoxin proved to be protein with a molecular weight of 140000 daltons and the isoelectric point equal to 4.4 . The biological activity of S . typhimurium enterotoxin was neutralized with homologous antiserum and with antiserum to cholera enterotoxin . Heating the preparation at 75 degrees C for 30 minutes led to a considerable decrease in its enterotoxic activity. Res Vet Sci, 1984 Sep, 37(2), 230 - 3 Growth of Salmonella typhimurium in the caecum of gnotobiotic chickens with Eimeria tenella; Fukata T et al.; Gnotobiotic chickens infected with Eimeria tenella (5 X 10(4) oocysts per bird) received an oral inoculation of 100 Salmonella typhimurium two, four, six or eight days after coccidial infection at four days old . When S typhimurium was given two or four days after E tenella infection, S typhimurium counts in the caecal contents were similar to the counts in birds infected with S typhimurium alone . When S typhimurium was given six or eight days after E tenella infection, counts of the organism were significantly greater than with S typhimurium infection alone . There were no differences in the number of chickens positive for S typhimurium in the caecal contents, bile, liver and spleen between the two groups. J Gen Microbiol, 1984 Sep, 130 ( Pt 9), 2277 - 83 Mechanism of the protective action of anti-Salmonella IgM in experimental mouse salmonellosis; Saxen H; The kinetics of mouse salmonellosis caused by Salmonella typhimurium was studied in mice preinjected with the IgM or IgG fraction prepared from a rabbit anti-Salmonella serum . Compared on the basis of antibody units determined by an enzyme immunoassay, IgM was ten times more effective than IgG in promoting removal of the bacteria from blood after intravenous (IV) injection and their uptake in the reticuloendothelial system (RES) . The subsequent killing of the bacteria was, however, only minor, in accord with the negligible protective effect of serum antibodies in IV infection . IgM was over 1000 times more effective than IgG in promoting killing of the bacteria after intraperitoneal (IP) challenge . Neither antibody had an effect on the multiplication of the bacteria in the RES . The protective action of antibody was thus almost entirely mediated by peritoneal-cavity cells acting in the very early phase of infection . The greater effect of IgM is suggested to be a special feature of Salmonella infections, connected with the capacity of these bacteria for intracellular survival and multiplication in the RES. Am J Vet Res, 1984 Sep, 45(9), 1858 - 61 Vaccination of calves against Salmonella dublin with aromatic-dependent Salmonella typhimurium; Smith BP et al.; Ten Holstein calves were divided into 2 groups . Five calves served as nonvaccinated controls, and 5 calves were vaccinated IM at 2 and 3 weeks of age with 10(9) aromatic-dependent (aro-) Salmonella typhimurium strain SL1479 containing O antigens 1, 4, 12 . Serious adverse reactions to vaccination were not observed in the calves . Mean maximum rectal temperature increase in the vaccinated calves was 1.5 C . One calf had diarrhea and depressed appetite for 1 day after vaccination . At 5 weeks of age, all calves were challenge exposed orally with 1.5 X 10(11) virulent S dublin strain SL1367 (O antigens 9,12) . After challenge-exposure inoculum was given, 1 of 5 vaccinated calves died and 4 of the 5 nonvaccinated calves died (P less than 0.05) . Thus, some cross serotype protection against S dublin was induced by parenteral vaccination of calves with aro- S typhimurium strain SL1479, although protection was not complete. J UOEH, 1984 Sep 1, 6(3), 257 - 63 {Mutagenesis of amino and/or methyl analogs of acridine in Salmonella and yeast: a comparison between the spot-test and the pre-incubation methods}; Goto C et al.; The relationship between mutagenic activities and chemical structure of acridine derivatives was examined by using Salmonella typhimurium strains TA 1537 and TA 1977 and yeast Saccharomyces cerevisiae . Most analogs with amino and/or methyl group(s) caused frameshift-type mutation in Salmonella without mammalian microsomal enzyme activation . The 9-amino analogs were strong mutagens and mutagenicity was also increased when 10-position was methylated in 1-amino and 2-amino compounds . However, 9-amino derivatives did not cause mitochondrial mutation in yeast, where 3,6-diamino and/or 10-methyl groups were structural requisites for significant mutagenic activity . In comparison with the pre-incubation method, the spot-test method was shown to be less sensitive . The mutagenicity of some compounds could not be detected by the spot-test method . Mutagenic activities of these drugs were revealed and increased markedly with an increase in pre-incubation period up to 20 min, suggesting that the pre-incubation of tester bacteria with a compound prior to a mutagenicity assay is necessary for the detection of mutagenesis of these compounds. Food Chem Toxicol, 1984 Sep, 22(9), 725 - 30 Effects of vitamin A on cyclophosphamide mutagenicity in vitro (Ames test) and in vivo (mouse micronucleus test); Busk L et al.; The effect of vitamin A on cyclophosphamide mutagenicity was measured both in vitro and in vivo . In the Ames test in Salmonella typhimurium TA1535 with mouse-liver S-9 mix, the addition of retinol, retinyl acetate or retinyl palmitate caused a dose-dependent inhibition of cyclophosphamide mutagenicity . In the micronucleus test in male NMRI mice fed low, normal or high levels of vitamin A, the induction of micronuclei in bone marrow by an ip dose of cyclophosphamide was unaffected by vitamin A status . Thus, this study provides no evidence that activation of a procarcinogen in the liver or bone marrow of mice can be modified by vitamin A . One of the possible reasons for the observed absence of inhibition by vitamin A in vivo may be a lack of correlation between the oral dose of retinoid and the resulting level of vitamin A in the bone marrow . The difference between results in vitro and in vivo may also have been due to a difference in the availability and potency of added vitamin A in vitro compared with the forms absorbed and stored in vivo. Poult Sci, 1984 Sep, 63(9), 1732 - 7 Effect of Eimeria tenella infection in chickens fed the feed artificially contaminated with Salmonella typhimurium; Morishima H et al.; Effect of Eimeria tenella infection on Salmonella typhimurium infection of chickens was tested using feed experimentally contaminated with S . typhimurium . Four experiments were conducted . In Experiments 1 and 2, chickens received feed contaminated with 10(3) or 10(2) cfu of S . typhimurium per g for 5 days after E . tenella infection . In Experiments 3 and 4, chickens were fed feed contaminated with 10(2) or 10 cfu of S . typhimurium per g for 3 days before E . tenella infection . In all experiments, chickens were necropsied 3 to 14 days after E . tenella infection . The number of S . typhimurium in the cecal contents was counted and the presence of the organism in the liver and bile was examined . In Experiments 1 and 2, there was no significant difference in S . typhimurium infection between the group infected with S . typhimurium alone and the group infected with both E . tenella and S . typhimurium . In Experiments 3 and 4, S . typhimurium counts in the cecal contents of chickens in the concurrently infected group were significantly greater than those of chickens in the S . typhimurium alone-infected group. Mutat Res, 1984 Sep, 141(1), 11 - 4 The direct mutagenic activity of alpha, omega-dihalogenoalkanes in Salmonella typhimurium . Strong correlation between chemical properties and mutagenic activity; Buijs W et al.; A series of 18 alpha, omega-dihalogenoalkanes (kappa(CH2)n kappa with n = 1-6 and kappa = Cl, Br, I) was tested for direct mutagenic activity in Salmonella strains TA1530, TA1535 and TA100 using spot-test procedures . The results indicate that the mutagenic behaviour of these compounds is strongly dependent upon the carbon chain length as well as the type of halogen involved . This behaviour correlates with the leaving group ability and the degree of neighbouring group participation in nucleophilic displacement reactions of the different halogen atoms. J Bacteriol, 1984 Sep, 159(3), 900 - 4 Heterogeneity of lipid A: comparison of lipid A types from different gram-negative bacteria; Mattsby-Baltzer I et al.; Chloroform-soluble purified lipid A preparations from 10 sources, including five Escherichia coli strains (EH100, K-12, O127, O111, RCDC), two Salmonella strains (Salmonella typhimurium, Salmonella minnesota R595), Shigella sonnei II, and a hybrid of Shigella flexneri and E . coli K-12, were compared with lipid A from S . flexneri . Purified lipid A from S . flexneri was earlier found to be composed of eight fractions . The various lipid A preparations were assayed by thin-layer chromatography . Chromatograms were stained for phosphate or carbohydrate by molybdenum blue or orcinol, respectively . The number of major bands found for each lipid A preparation varied between 7 and 10, depending on the source . Comparable bands, based on Rf, were found among all of the different lipid A preparations, but the quantity of each band varied between the sources of lipid A . Four bands (designated 2, 3, 7, and 8) were abundant in every preparation . Variations of conditions used for preparing lipid A, such as changing of hydrolysis time, did not affect the appearance of lipid A on thin-layer chromatography . Change in the type of acid used for hydrolysis also did not affect the band pattern, but it did change the quantitative amounts of the various bands to some degree. J Bacteriol, 1984 Sep, 159(3), 1090 - 2 Map locations and functions of Salmonella typhimurium men genes; Kwan HS et al.; Menaquinone (men) mutants of Salmonella typhimurium isolated on the basis of their inability to produce trimethylamine were characterized with respect to mutation site, the ability to cross-feed each other and be cross-fed by known Escherichia coli men mutants, and response to intermediates of the menaquinone biosynthetic pathway . Cross-feeding tests were based on the requirement of menaquinone for hydrogen sulfide production . Genotypes corresponding to the menA, B, C, D, and possibly E genes described in E . coli were all identified . Additional studies of deletions in the menBCD area revealed that this cluster lies between ack/pta and glpT, as in E . coli . The ack and pta mutants were also defective in the production of trimethylamine and failed to produce gas in the absence of added formate. J Bacteriol, 1984 Sep, 159(3), 1056 - 9 Excretion of unassembled flagellin by Salmonella typhimurium mutants deficient in hook-associated proteins; Homma M et al.; Of the flagellar filamentless mutants of Salmonella typhimurium, the flaV, flaU, and flaW mutants, which are defective in hook-associated proteins, synthesized flagellin molecules, but flagella did not polymerize at the tips of the mutant hooks and were excreted into the culture medium as intact monomers. Radiat Res, 1984 Sep, 99(3), 609 - 26 Toxic variability and radiation sensitization by Pt(II) analogs in Salmonella typhimurium cells; Richmond RC et al.; A rationale is presented for the development of toxic, i.e., cytocidal, antitumor drugs as clinical hypoxic cell radiation sensitizers . Pt(II) complex-induced hypoxic cell radiation sensitization may occur from Pt(II) complex in free solution and Pt(II) bound to DNA . Although both the free solution and the bound compartments may operate, the free solution compartment is more likely amenable to experimental and clinical control in the case of systemically active Pt drugs . Assuming equivalent cell uptake of different Pt(II) complexes, the free solution compartment of Pt(II) sensitization can be increased by utilizing less toxic analogs of the antitumor drug cis-dichlorodiammineplatinum(II) . One such less toxic Pt(II) sensitizer currently in clinical use is found to be cis-(1,1-cyclobutanedicarboxylato)diammineplatinum(II) . A new finding of both clinical and mechanistic usefulness is described: irradiation of hypoxic solutions of four cis-Pt(II) complexes, but not two trans-Pt(II) complexes, creates products that cause toxicity in excess of the unirradiated solutions. Radiat Res, 1984 Sep, 99(3), 596 - 608 Toxic variability and radiation sensitization by dichlorodiammineplatinum(II) complexes in Salmonella typhimurium cells; Richmond RC; The oxidative coordination compound cis-dichlorodiammineplatinum(II) (cis-DDP) is again shown to be a hypoxic cell radiation sensitizer . The mechanism of cis-DDP-induced radiation sensitization is complex . Results here indicate that cis-DDP sensitization operates in part through reactive free radicals, in part through the interactions of radiation-induced reactive Pt(I) intermediates, and in part through the involvement of thermodynamic and kinetic aspects of Pt(II)-DNA binding during irradiation . For the first time, radiation sensitization by trans-DDP is compared with a sensitizing concentration of cis-DDP within the same study . Both analogs are sensitizers, but with significant differences . Further, irradiated hypoxic solutions of cis-DDP are found to be more toxic than unirradiated solutions. J Med Chem, 1984 Sep, 27(9), 1156 - 61 Synthesis, spectral analysis, and mutagenicity of 1-, 3-, and 6-nitrobenzo{a}pyrene; Chou MW et al.; The mutagenic environmental pollutants 1-, 3-, and 6-nitrobenzo{a}pyrene were synthesized . Nitration of 7,8,9,10-tetrahydrobenzo{a}pyrene with sodium nitrate in trifluoroacetic acid and acetic anhydride at ambient temperature gave a mixture of 1-, 3-, and 6-nitro-7,8,9,10-tetrahydrobenzo{a}pyrene, which was separated by chromatography . Dehydrogenation of the isolated nitrotetrahydrobenzo{a}pyrenes with 2,3-dichloro-4,5-dicyano-1,6-benzoquinone produced 1-, 3-, and 6-nitrobenzo{a}pyrene in high yield . Comparison of the spectral data of these compounds with those obtained from direct nitration of benzo{a}pyrene confirmed that 1- and 3-nitrobenzo{a}pyrenes are indeed the minor products of the latter reaction . This confirmation also verifies that 1- and 3-nitrobenzo{a}pyrene were the minor nitrated products of benzo{a}pyrene formed in model air atmospheres . The 1-, 3-, and 6-nitrobenzo{a}pyrene were mutagenic in Salmonella typhimurium tester strains TA98 and TA100 in the presence of a mammalian microsomal (S9) activating system . Both 1- and 3-nitrobenzo{a}pyrene, but not 6-nitrobenzo{a}pyrene, were also direct-acting mutagens in these strains . However, only 6-nitrobenzo{a}pyrene exhibited weak mutagenic activity when tested in Chinese hamster ovary cells, while only 3-nitrobenzo{a}pyrene produced a concentration-dependent decrease in cellular survival. Cell Immunol, 1984 Sep, 87(2), 528 - 37 Separate transfer of mouse protection and delayed-type hypersensitivity with Salmonella typhimurium transfer factor; Kita E et al.; Delayed-type hypersensitivity (DTH) induced with Salmonella typhimurium transfer factor (TF) contributed to an increase in mean survival days of mice challenged with homologous organisms and afforded only a low level of host protection as determined by survival rate, compared with that obtained by active immunization . TF of other enteric bacteria could transfer DTH which is cross-reactive to salmonella antigen but did not afford host protection . Although TF of Listeria monocytogenes did not transfer the cross-reactive DTH, it could confer the significant increase in mean survival days against the lethal challenge with S . typhimurium . Listerial ribosomal vaccine conferred the high level of mouse protection without inducing DTH to salmonella antigen . The resistance generated upon active immunization with listerial ribosomal vaccine could be enhanced by the injection of S . typhimurium TF to the same level as that obtained after immunization with homologous ribosomal vaccine . Among salmonella TF, there could be no cross-reactive immunity between S . typhimurium and S . choleraesuis, although the cross-reactive DTH was observed . The DTH transfer ability of TF was sensitive to Pronase which could not affect the ability to transfer host immunity, but RNase could abolish the ability to transfer host immunity without impairing DTH transfer activity . These results suggest that in mouse typhoid infection, DTH is not associated with host protection as determined by survival rate. J Immunol, 1984 Sep, 133(3), 1190 - 6 Differences in delayed-type hypersensitivity responses in various mouse strains in the C3H lineage infected with Salmonella typhimurium, strain SL3235; Killar LM et al.; Immunization with a virulent Salmonella typhimurium, strain SL3235, has been found to provide high levels of protection against challenge with virulent Salmonella in hypersusceptible mouse strains in the C3H lineage . These mouse strains include the lipopolysaccharide-hyporesponsive C3/HeJ mouse and the closely related but lipopolysaccharide-responsive C3HeB/FeJ mouse . To assess the role of cellular immunity in the protection elicited by this attentuated organism, delayed-type hypersensitivity (DTH) was measured in these mouse strains and in inherently resistant mice . Of the mouse strains tested, only the inherently resistant CD-1 and C3H/HeNCrlBR mice developed significant DTH responses, as assessed by footpad swelling tested at various times after immunization with SL3235 . The hypersusceptible C3H/HeJ and C3HeB/FeJ mice failed to exhibit significant DTH responses despite their high levels of immunity. Chem Biol Interact, 1984 Sep 1, 51(1), 49 - 62 Mutagenic activity of possible metabolites of 4-nitrobiphenyl ether; Miyauchi M et al.; A series of possible metabolites--4-nitrosobiphenyl ether (4-NO), 4-hydroxylaminobiphenyl ether (4-NHOH), 4-aminobiphenyl ether (4-NH2), 4-hydroxyacetylaminobiphenyl ether (4-N(OH)Ac), 4-acetoxyacetylaminobiphenyl ether (4-N(OAc)Ac)involved in the toxic effects of 4-nitrobiphenyl ether (4-NO2) was synthesized and tested for mutagenic activity toward Salmonella typhimurium TA100 strain in the presence and the absence of liver homogenates of guinea pig treated with Kaneclor-500 . 4-NO2, 4-NO and 4-NHOH showed direct-acting mutagenicity . 4-NO and 4-NHOH showed high mutagenic activity, while the mutagenic activity of 4-NO2 was very weak compared to 4-NO and 4-NHOH . 4-NO showed antimicrobial action at high concentrations . The other three compounds tested induced no mutation . Upon addition of NAD(P)H, the mutagenic activities of 4-NO and 4-NHOH were slightly enhanced, but no enhancement was observed by addition of NAD(P)+ . Metabolic activation with guinea pig liver homogenates enhanced the mutagenic activities of 4-NO2 and 4-NO, and converted 4-NH2, 4-N(OH)Ac and 4-N(OAc)Ac to the product(s) responsible for the mutagenic activity . Addition of bis(p-nitrophenyl)phosphate, a deacetylase inhibitor, inhibited the mutagenic activities of 4-N(OH)Ac and 4-N(OAc)Ac by about 70% in the presence of NADPH and about 77% in the absence of NADPH . High performance liquid chromatography (HPLC) analysis of non-enzymatic conversion-products of 4-NHOH and 4-BO with and without NADPH indicated that 4-NHOH disappeared after 30 min of incubation and was converted completely to 4-NO without NADPH, while with NADPH, 4-NHOH disappeared very slowly and was detected even after 4 h of incubation . In the case of 4-NO, no decrease of 4-NO was observed without NADPH, while with NADPH 4-NO decreased quickly and a significant amount of 4-NHOH appeared . The mechanism of the NAD(P)H-dependent increase in mutagenicity is also discussed. J Pediatr Gastroenterol Nutr, 1984 Sep, 3(4), 585 - 92 Studies with enterotoxigenic microorganisms: effects of candidate antidiarrhoeals in experimental animals in vivo; Burke V et al.; Chlorpromazine or aspirin alone, when given to rats parenterally, reduced intestinal fluid secretion induced by cell-free preparations of enterotoxigenic organisms including Escherichia coli, Aeromonas hydrophila, Staphylococcus pyogenes, and Salmonella typhimurium . A combination of chlorpromazine-aspirin given parenterally caused much more marked reduction of fluid secretion . Indomethacin also had significant antisecretory effects against a range of bacterial enterotoxins, while loperamide was effective against heat-labile toxin (LT)-positive E . coli and A . hydrophila . Nicotinamide increased net fluid absorption in the presence of E . coli LT, A . hydrophila, and S . typhimurium . Of the adsorbents tested, aluminum hydroxide showed a positive effect only with E . coli LT and A . hydrophila, while cholestyramine affected net fluid flux only with E . coli ST (heat-stable toxin) . Charcoal was effective against all microorganisms tested but only when premixed with the perfusate before the experiments . Aspirin and chlorpromazine probably act at multiple sites to decrease intestinal secretion, and the combination of low doses of these drugs with possibly different sites of action may have advantages over a single agent used in high dosage. Genetics, 1984 Sep, 108(1), 1 - 23 Functional interchangeability of DNA replication genes in Salmonella typhimurium and Escherichia coli demonstrated by a general complementation procedure; Maurer R et al.; Twenty-four genes from Salmonella typhimurium that affect DNA replication were isolated from a lambda-Salmonella genomic library by lysogenic complementation of temperature-sensitive mutants of Salmonella or E . coli, using a new plaque complementation assay . The complementing lambda clones, which make red plaques in this assay, and noncomplementing mutant derivatives, which make uncolored plaques, were used to further characterize the temperature-sensitive Salmonella mutants and to establish the functional similarity of E . coli and Salmonella DNA replication genes . For 17 of 18 E . coli mutants representing distinct loci, a Salmonella gene that complemented the mutant was found . This result indicates that single Salmonella replication proteins are able to function in otherwise all E . coli replication complexes and suggests that the detailed properties of Salmonella and E . coli replication proteins are very similar . The other seven Salmonella genes that were cloned were unrelated functionally to any E . coli genes examined . --As an aid to the derivation of chromosomal mutations affecting some of the cloned genes, a general method was developed for placing a transposon in the Salmonella chromosome in a segment corresponding to cloned DNA . Chromosomal mutations were derived in Salmonella affecting a gene (dnaA) that was cloned by complementation of an E . coli mutant by using the transposon-encoded drug resistance as a selectable marker in local mutagenesis. J Infect Dis, 1984 Sep, 150(3), 425 - 35 Immunity to infection with Salmonella typhimurium: mouse-strain differences in vaccine- and serum-mediated protection; Eisenstein TK et al.; Three mouse strains in the C3H lineage--C3H/HeJ, C3HeB/FeJ, and C3H/HeNCr1BR--were tested for their ability to be protected against infection with Salmonella typhimurium by a panel of nonviable vaccines and by passive transfer of hyperimmune serum . These strains differ in their innate susceptibilities to infection with S . typhimurium, but all are histocompatible . The same vaccines showed a widely different ability to protect different mouse strains . Ability to protect was not closely related to the capacity of the mice to make either agglutinating or anti-O antibody (as shown by ELISA) in response to a particular vaccine . Passive transfer of antibody was shown to protect inherently resistant mice but not inherently susceptible strains . These observations suggest that reported discrepancies in vaccine efficacy among laboratories may be attributable to differences in the mouse strains used and raise the question as to what might be an appropriate mouse model for human infections with Salmonella species. J Antimicrob Chemother, 1984 Sep, 14 Suppl B, 153 - 9 Severe multiresistant Salmonella typhimurium systemic infections in Central Africa--clinical features and treatment in a paediatric department; Lepage P et al.; During a 21-month period, we observed an outbreak of severe systemic infections due to multiresistant Salmonella typhimurium among 66 children in the in-patient Department of Paediatrics of Kigali, Rwanda . These infections were more likely to occur in subjects who had stayed for a long time in the hospital for severe illness and/or malnutrition . The children usually presented first with mild to moderate diarrhoea and fever . Later, sever pulmonary involvement was often noted (rales: 58%; respiratory distress: 42%) . Moreover, there were four cases of abscess, three arthritis and one meningitis . Of the 66 children, 48 were treated with cefotaxime . The fatality-rate among this group was 10.4% . The fatality-rate among the 18 other untreated patients was 77.9%, suggesting a high efficiency of cefotaxime against these strains of multiresistant Salm . typhimurium. J Bacteriol, 1984 Sep, 159(3), 951 - 7 Regulation of Salmonella typhimurium ilvYC genes; Blazey DL et al.; The Salmonella typhimurium LT2 ilvYC genes were studied by fusion of each gene to the Escherichia coli K-12 galK gene . The expression of ilvY and ilvC could then be determined by measurement of the galK-encoded galactokinase enzyme . The promoter for ilvC, pC, was located by this technique to a 0.42-kilobase BglII-EcoRI fragment of the S . typhimurium ilvGEDAYC gene cluster . This sequence was completely sufficient for alpha-acetohydroxyacid-inducible galK expression . The ilvY gene was located within a 1.0-kilobase XhoI-SalI fragment . ilvY gene expression was constitutive with respect to ilv-specific control signals . The ilvY gene was transcribed in the same direction as the other two transcriptional units in the ilvGEDAYC gene cluster, ilvGEDA and ilvC . Transcription of the ilvC gene was completely dependent upon the activity of its own promoter, pC, and independent from transcription of the ilvY gene . The role of the intervening region between ilvY and ilvC in regulation of ilvC expression was explored. J Bacteriol, 1984 Sep, 159(3), 1000 - 5 Analysis of promoter mutations in the histidine transport operon of Salmonella typhimurium: use of hybrid M13 bacteriophages for cloning, transformation, and sequencing; Lee GS et al.; Mutations that cause an increased level of expression of the histidine transport operon were isolated and characterized genetically . Five independently isolated promoter-up mutations were transferred to an M13 hybrid phage that carries the histidine transport operon, and their nucleotide sequences were determined . For all five mutations, the change was the same as the one previously determined for promoter-up mutation dhuA1: a C-to-T change in the Pribnow box rendered this region more homologous to the consensus sequence . Methods for enabling Salmonella typhimurium to support growth of M13 phage effectively and for easy transfer of chromosomal mutations onto the hybrid phage are presented. Genetics, 1984 Sep, 108(1), 25 - 38 Genetic analysis of DNA replication in bacteria: dnaB mutations that suppress dnaC mutations and dnaQ mutations that suppress dnaE mutations in Salmonella typhimurium; Maurer R et al.; We have isolated and characterized extragenic suppressors of mutations in two different target genes that affect DNA replication in Salmonella typhimurium . Both the target and the suppressor genes are functional homologues of known replication genes of E . coli that were identified in intergeneric complementation tests . Our results point to interactions in vivo involving the dnaB and dnaC proteins in one case and the dnaQ and dnaE proteins in the other case . The suppressor mutations, which were isolated as derivatives of lambda-Salmonella in vitro recombinants, were detected by an adaptation of the red plaque complementation assay . This method was applicable even when the locus of suppressor mutations was not chosen in advance. Med J Aust, 1984 Aug 18, 141(4), 217 - 9 Salmonella food poisoning; Keogh AM et al.; During an outbreak of Salmonella typhimurium food poisoning in September 1983, in Sydney, 10 affected subjects were admitted to the same hospital . On admission to hospital, all patients were severely dehydrated; two patients developed acute tubular necrosis, while a third patient had myopericarditis . Bacteraemia was confirmed in two patients, in one of whom no organisms were isolated from stool cultures . Antibiotic agents were administered to all patients, because of the unusually severe nature of the infection . This outbreak illustrates that salmonella gastroenteritis, although usually fairly mild and self-limiting, can be a virulent disease resulting in serious complications . Appropriate management should include careful initial assessment of suspected cases, vigorous correction of fluid and electrolyte disturbances, and judicious use of antibiotic agents when bacteraemia or severe toxaemic features are present. Mutat Res, 1984 Aug, 130(4), 267 - 72 Depletion of the reduced glutathione level in the liver and production of the mutagens in the intestine in the mice inducing hepatoma by feeding on a high level dose of sorbic acid; Tsuchiya T et al.; Mutagenicity was detected using Salmonella typhimurium TA98 in the acidic components of the intestinal contents of mice, in which a high incidence of hepatoma had been reported due to feeding on a diet containing 15% sorbic acid (Ishizawa et al., 1980) . Furthermore, the glutathione level in the liver of the 15% sorbic acid group was decreased to 40% of the amount found in controls after a 3-month feeding period, and this low level was maintained for long periods (12 months) until the end of the experiments . There was also a close correlation between the extent of depletion of the hepato-glutathione level and the concentration of sorbic acid added to the diet . Consequently, the hepatoma which developed in mice fed a 15% sorbic acid diet was considered to be induced both by the depletion of the hepato-glutathione level over the long periods, and by the gradual production of various mutagens in the intestine which were absorbed and transferred to liver to be metabolically activated. Infect Immun, 1984 Aug, 45(2), 332 - 8 Zinc concentration and survival in rats infected with Salmonella typhimurium; Tocco-Bradley R et al.; Percent survival was measured in male rats injected intravenously with live Salmonella typhimurium when plasma and tissue zinc levels were manipulated . Alzet pumps implanted intraperitoneally infused zinc gluconate or sodium gluconate (controls) from the onset of infection to 72 h postinfection . Plasma and tissue zinc levels were manipulated by infusing (i) 180 micrograms of Zn per h to achieve supranormal plasma and tissue zinc concentrations, (ii) 120 micrograms of Zn per h to prevent the infection-induced fall and to maintain plasma zinc levels at noninfection levels while raising tissue levels above that of infected controls, and (iii) 30 micrograms of Zn per h to increase tissue zinc levels while allowing the infection-induced decrease in plasma zinc . Preventing the fall in plasma zinc while raising liver zinc to supranormal levels enhanced rather than reduced percent survival; raising plasma and liver zinc to supranormal levels returned survival to control levels . Loading the liver with an excess of zinc without changing plasma zinc (30 micrograms of Zn per h) did not increase percent survival in the infected host . Pretreatment or administration of zinc at the time of infection led to increased percent survival compared with administration of zinc 4 h after the onset of infection. Genetika, 1984 Aug, 20(8), 1270 - 8 {Genetic activity of base analogs in Salmonella typhimurium and Escherichia coli and Saccharomyces cerevisiae}; Pavlov IuI et al.; The study of 6-N-hydroxylaminopurine (HAP) and 2-amino-6-N-hydroxylaminopurine (AHAP) activity in bacteria and the yeast was undertaken . AHAP was found to be more effective as a mutagen in bacteria and HAP--in the yeast . Mutagenic and lethal effects or analogues were independent of excision and mutagenic repair both in bacteria and the yeast . Deletion in uvrB region of Salmonella genome leads to hypersensitivity to lethal and mutagenic action of analogues . Both of the latter only cause reversions of base-substitution but not frameshift mutations . Considering the data obtained and the information from published papers, we proposed that HAP and AHAP exert their mutagenic action, like classical analogues, by means of incorporation into DNA and disturbing the regular replication laws. Mutat Res, 1984 Aug-Sep, 137(2-3), 79 - 88 Lack of mutagenicity of diphenylhydantoin in in vitro short-term tests; Leonard A et al.; The mutagenicity of diphenylhydantoin (DPH) and its major metabolite, 5-(p-hydroxyphenyl)-5-phenylhydantoin (HPPH), has been re-evaluated by the Ames test using Salmonella typhimurium and, for DPH only, by an in vitro cytogenetic test with human lymphocytes and a turbidimetric assay of tubulin polymerization . As negative results were obtained in all test systems used here, one has to conclude that DPH is devoid of mutagenic properties. Toxicology, 1984 Aug, 32(2), 117 - 30 Metabolism, DNA binding, and cytotoxicity of aflatoxin B1 in tracheal explants from Syrian hamster; Coulombe RA Jr et al.; Metabolism, DNA binding and cytopathological effects of aflatoxin B1 (AFB1) were studied in isolated tracheal explants from Syrian golden hamsters . Explants were exposed to 0.1, 0.5 and 1.0 microM {14C}AFB1 in Dulbeccos's modified Eagle medium for 24 h, then analyzed for AFB1-DNA binding and AFB1 metabolism . Binding (pmol AFB1/mg DNA) was dose-related (16.3 +/- 1.9 to 180.8 +/- 16.1) and analysis of the culture medium revealed the metabolic conversion products aflatoxicol (AFL) and aflatoxin Q1 (AFQ1) . Ultrastructural analysis of sections of tracheal epithelium revealed degenerative changes primarily in the non-ciliated epithelial cells . Autoradiographic analysis of the same treated explants, however, showed no discernible distribution of label with respect to either cell type or cell location, with the exception of increased grain densities overlying vacuoles containing dark droplets . In addition, S9 prepared from hamster trachea was shown to activate AFB1 to mutagens detectable by Salmonella typhimurium TA 98, but was approximately 70 times less active on a per mg protein basis than was S9 prepared from hamster liver . These results demonstrate the metabolic capabilities of tracheal epithelial cells in the activation of AFB1, thus indicating that AFB1 present in respiratory particles may be activated by pulmonary mixed-function oxidases, posing a hazard to those exposed. J Environ Sci Health B, 1984 Aug, 19(6), 565 - 77 Genotoxicity of methyl parathion in short-term bacterial test systems; Rashid KA et al.; Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA-damage . Methyl parathion was mutagenic to S . typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes . In DNA repair tests, methyl parathion was effective in inducing damage to the S . typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms . Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1984 Aug, 257(3), 414 - 25 {Protection against experimental Salmonella typhimurium infection in mice . Immunostimulating activity of heterologous Salmonella S-forms, R-mutants, lipopolysaccharides and muramyl dipeptide in vaccines combined with S . typhimurium}; Schlecht S; In vaccines consisting of acetone-killed Salmonella, 90% of the bacteria were replaced by: heterologous Salmonella S-forms, R-mutants of Salmonella or Escherichia coli, lipopolysaccharide from S . typhimurium (S-form) or from R-mutants of Salmonella or E . coli and by muramyl dipeptide . Active immunizations of NMRI mice with these vaccines were undertaken . Mice received two intraperitoneal injections of the vaccines at intervals of 14 days, and were challenged with various doses of S . typhimurium C5 10 days later . The protective capacity of the mixed vaccines was compared with that of monovaccines (S . typhimurium) and with the effectiveness of vaccines consisting of the supplementing component (relatively weak immunizing ability) alone . The LD50 served as criterion for protective capacity . The results showed that S . typhimurium S-form could be replaced by Salmonella R-mutants belonging to different chemotypes without a recognizable decrease in protective immunizing capacity of the vaccines . Effective vaccines were also attained when heterologous Salmonella S-forms, which exhibit no O-antigenic determinants in common with S . typhimurium, were used as supplements . Mixtures with E . coli R-mutants proved to be less effective . In addition, the diminished dose of S . typhimurium could be so effectively supplemented with lipopolysaccharide from S . minnesota R 595 that complete protection was achieved . In contrast, comparable doses of R lipopolysaccharides from E . coli were somewhat less effective . Vaccines of LPS-extracted bacteria exhibited a reduced protective capacity and were ineffective when used as supplements in mixed vaccines . Analogous results were obtained when R-mutants served as basic vaccines in place of the S . typhimurium S-form indicating that an immunogenic component is enhanced in these organisms too and that their immunogenic capacity is not brought about barely by general stimulation of the immune system . However, the full effectiveness of the R-monovaccines was often not attained with supplements . Immunization with mixed vaccines of S . typhimurium S-form and heterologous Salmonella S-forms or Salmonella R-mutants led to equally high agglutinin and haemagglutinin titers as those obtained with monovaccines of S . typhimurium S-form. Sci Total Environ, 1984 Aug 1, 37(2-3), 171 - 6 Chlorination of ozonated soil fulvic acid: mutagenicity studies in Salmonella; Kowbel DJ et al.; Samples of soil fulvic acid (SFA) were ozonated and subsequently chlorinated under acidic or slightly basic conditions . The residues were tested for His+ reversion in a fluctuation assay, using Salmonella typhimurium TA100 as the tester strain . The ozonated/chlorinated samples were mutagenic, but activity was dependent on the amount of ozone utilized and the pH of the reaction medium . Although increases in cell concentrations were also induced by some mutagenic samples, this alone did not account for the mutagenicity observed . Unchlorinated samples displayed insignificant activity. Mutat Res, 1984 Aug-Sep, 137(2-3), 89 - 93 Mutagenicity of hexachlorobutadiene, perchlorobutenoic acid and perchlorobutenoic acid chloride; Reichert D et al.; Hexachloro(1,3)butadiene (HCBD) is a well known environmental contaminant . The nephrocarcinogenic potential of HCBD has been shown in long-term studies with rats . Experiments were performed to assist in determining whether this effect is mediated by epigenetic or genotoxic mechanisms and to compare the mutagenic properties of HCBD with those of its monooxidation products, perchloro-3-butenoic acid (PCBA) and perchloro-3-butenoic acid chloride (PCBAC), which are conceivable metabolites of HCBD . All 3 compounds are mutagenic to the Salmonella typhimurium tester strain TA100 . The mutagenic effect is dose-dependent and parallels the chemical reactivity of the compounds . HCBD is only mutagenic in the presence of drug-metabolizing enzymes (S9 mix) with an increased protein content . The mutagenic response after incubation with PCBAC and PCBA is 2-3-fold that of HCBD . Additionally, both PCBAC and PCBA exert a mutagenic response in the absence of S9 mix . The experiments support the assumption of a genotoxic potential of HCBD. Mutat Res, 1984 Aug-Sep, 137(2-3), 71 - 8 The mutagenicity on Salmonella typhimurium of nitrobenzoic acids and other wastewater components generated in the production of nitrobenzoic acids and nitrotoluenes; Sundvall A et al.; The wastewater contained mutagens which induced mutations in Salmonella typhimurium TA1535, TA1538, TA98 and TA100 . By the use of nitroreductase-proficient and -deficient tester strains, it was possible to demonstrate that the mutagens were to a great extent aromatic nitro compounds . 30-40% of the mutagenicity could be related to the 16 identified nitroaromatic compounds . Although 13 of these induced mutations, one single compound, 3,5-dinitrobenzoic acid, was responsible for more than 80% of their total mutagenicity . p-Nitrobenzoic acid was used for further studies of the enzymatic nitroreduction leading to the formation of reactive intermediates . The bacterial enzymes and the active metabolites did not seem to be oxygen-sensitive, as the mutagenicity was decreased when anaerobic incubation was applied . The addition of dicoumarol resulted in a decreased effect, indicating that bacterial DT diaphorase or an enzyme with similar properties is responsible at least in part for the activation of this compound . Under our experimental conditions rat-liver enzymes were not able to produce any detectable amounts of mutagenic metabolites of p-nitrobenzoic acid when the nitroreductase-deficient strain TA100NR was used. Mutat Res, 1984 Aug-Sep, 137(2-3), 133 - 7 Bacterial reversion assay and micronucleus test carried out on hydrogenated glucose syrups 'Malti-Towa' (powder) and maltitol crystal; Takizawa Y et al.; Two preparations of maltitol (4-O-alpha-D-glucopyranosyl-D-sorbitol), hydrogenated glucose syrups and maltitol crystal, were examined for genotoxic potential by a battery of short-term tests . In the bacterial reversion assay, maltitol induced no detectable revertants in any of the tester strains, Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538, or Escherichia coli WP2/pKM101 at doses of 0.5-50 mg per plate with and without rat liver S9 mix . In the micronucleus test, no significant increase in the frequency of micronucleated erythrocytes was observed in bone marrow of mice after administration of the two preparations at 3.75-30 g per kg by gastric intubation. J Infect Dis, 1984 Aug, 150(2), 236 - 41 The significance of hospitals as reservoirs for endemic multiresistant Salmonella typhimurium causing infection in urban Brazilian children; Riley LW et al.; To identify possible sources of multiply drug-resistant Salmonella typhimurium among children in Sao Paulo, Brazil, we reviewed records of 470 children who had visited eight outpatient clinics from March 1981 to May 1982 and of 28 children who had been admitted to one referral hospital between June and November of 1982, and we examined plasmid profiles of the Salmonella isolates by agarose-gel electrophoresis . S . typhimurium was identified in 37 of these children . Case-control studies showed that children with S . typhimurium infections were more likely to have been hospitalized before onset of diarrhea than were either age-matched children without diarrhea (P = .031) or age-matched children with nonbacterial diarrhea (P = .035) . Four distinct plasmid profiles, each of which was temporally clustered, were identified in 20 (67%) of 30 S . typhimurium strains isolated from previously hospitalized children and in two (28%) of seven strains from children not previously hospitalized . Each of three of these four profiles was associated with a different hospital, results suggesting that multiresistant-S . typhimurium infections in Sao Paulo are often nosocomially acquired. Food Chem Toxicol, 1984 Aug, 22(8), 677 - 9 Evaluation of the genotoxicity of lac dye; Banerjee TS et al.; Red lac dye, a by-product of the shellac industry, has the potential for use in foods, drugs and cosmetics as a colouring agent . As part of a series of tests of the suitability of lac dye for this purpose an evaluation of its genotoxicity was carried out . Lac dye was non-mutagenic in Ames tests using five strains of Salmonella typhimurium with or without metabolic activation . No cytotoxicity or mutagenicity was observed in Chinese hamster lung (V79) cells exposed to lac dye in vitro . A clastogenic effect was observed in the bone-marrow cells of mice that had been treated with lac dye ip or orally. Toxicol Appl Pharmacol, 1984 Aug, 75(1), 137 - 46 Mutagenicity testing of agent orange components and related chemicals; Mortelmans K et al.; Components of the herbicide Agent Orange--2,4-dichlorophenoxyacetic acid (2,4,-D) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and their esters, and the contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)--and related chemicals were tested for mutagenicity using Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 . No mutagenic activity was observed for any of the chemicals tested. J Bacteriol, 1984 Aug, 159(2), 787 - 9 In vitro polymerization of flagellin excreted by a short-flagellum Salmonella typhimurium mutant; Ikeda T et al.; The culture medium of a short-flagellum mutant of Salmonella typhimurium contained a large amount of mutant flagellin and a small amount of strong inhibitors of flagellin polymerization . After being freed of the inhibitors, the mutant flagellin could be polymerized in vitro, although under nonphysiological conditions. J Bacteriol, 1984 Aug, 159(2), 704 - 12 New Salmonella typhimurium mutants with altered outer membrane permeability; Sukupolvi S et al.; We describe three new classes of Salmonella typhimurium mutants with increased sensitivity to hydrophobic agents . In contrast to many previously described mutants, the phage sensitivity pattern of these mutants did not give any indication of defective lipopolysaccharide . Furthermore, they had no detectable changes in their phospholipid or outer membrane protein composition, and their growth rate and cell morphology were normal . Class B mutants were nearly as sensitive to novobiocin, fusidic acid, erythromycin, rifampin, and clindamycin as are deep rough (heptoseless) mutants; in addition they were sensitive to methicillin, penicillin (to which heptoseless mutants are resistant), gentian violet, and anionic and cationic detergents . Class A and C mutants had less sensitive, but characteristic phenotypes . None of the three classes were sensitive to serum bactericidal action . The class B mutation mapped between map positions 7 and 11 on the S . typhimurium chromosome, and the class C mutation mapped between positions 5 and 7 . The map position for the class A mutation remained undefined, but it was separate from the class B and C mutations and, like those, did not correspond to any gene loci known to participate in the synthesis of major outer membrane constituents. J Bacteriol, 1984 Aug, 159(2), 663 - 7 Cytochrome o as a terminal oxidase and receptor for aerotaxis in Salmonella typhimurium; Laszlo DJ et al.; Cytochrome o was the only oxidase of the electron transport system that was present in exponentially growing Salmonella typhimurium ST1 . Identification of cytochrome o was made by the (CO-reduced)-minus-(reduced) difference spectra and by the photochemical action spectrum of the relief, by light, of CO-inhibited respiration . Cytochrome o also functioned as the receptor for chemotaxis to oxygen (aerotaxis) . The concentration of oxygen that elicits the maximum response for aerotaxis (0.7 microM) was similar to the Km for respiration (0.74 microM), and both aerotaxis and respiration were blocked 5 mM KCN. Cancer Res, 1984 Aug, 44(8), 3408 - 13 Identification of trans-1,2-dihydro-1,2-dihydroxy-6-nitrochrysene as a major mutagenic metabolite of 6-nitrochrysene; El-Bayoumy K et al.; Liver 9000 X g supernatant from rats was used to study the metabolism of {6- 14C}nitrochrysene under aerobic conditions . The major ethyl acetate-soluble metabolite (1.06 nmol/mg of protein in 30 min) was identified as 1,2-dihydro-1,2-dihydroxy-6-nitrochrysene, based on its mass, UV, and proton magnetic resonance spectra . Under aerobic conditions, 6-aminochrysene was not detected as a metabolite . However, when incubations were carried out in an atmosphere of 4% O2 in N2, both 1,2-dihydro-1,2-dihydroxy-6-nitrochrysene (0.04 nmol/mg of protein) and 6-aminochrysene (0.05 nmol/mg of protein) were detected . Further metabolism of the 14C-labeled 1,2-dihydro-1,2-dihydroxy-6-nitrochrysene by rat liver 9000 X g supernatant under aerobic conditions gave a major metabolite which was identified tentatively as 1,2-dihydroxy-6-nitrochrysene . The mutagenic activities of 6-nitrochrysene, trans-1,2-dihydro-1,2-dihydroxy-6-nitrochrysene and 6-aminochrysene were assessed in Salmonella typhimurium strains TA100 and TA98 . In the absence of rat liver 9000 X g supernatant, trans-1,2-dihydro-1,2-dihydroxy-6-nitrochrysene was the more potent mutagen in TA100 but, in TA98, it was less active than was 6-nitrochrysene . In the presence of rat liver 9000 X g supernatant, both trans-1,2-dihydro-1,2-dihydroxy-6-nitrochrysene and 6-nitrochrysene were more mutagenic in TA100 than in the assays performed without an activating system, and the dihydrodiol metabolite was more mutagenic than was 6-nitrochrysene . In TA98 with activation, trans-1,2-dihydro-1,2-dihydroxy-6-nitrochrysene, 6-aminochrysene, and 6-nitrochrysene were all mutagenic . The results of this study indicate that trans-1,2-dihydro-1,2-dihydroxy-6-nitrochrysene is a major proximate mutagen of 6-nitrochrysene in S . typhimurium TA100. J Immunol, 1984 Aug, 133(2), 988 - 92 IgA-dependent cell-mediated activity against enteropathogenic bacteria: distribution, specificity, and characterization of the effector cells; Tagliabue A et al.; Antibody-dependent cellular cytotoxicity (ADCC) against murine enteropathogenic bacteria such as Salmonella typhimurium and Salmonella tel aviv or Shigella X16 was assessed by using IgG, IgA, and secretory IgA (sIgA) in a 2-hr in vitro assay where peripheral and intestinal lymphocytes were used as effector cells . It was found that IgG could arm splenocytes (SpL) better than IgA . However, IgG did not arm lymphocytes from Peyer's patches (PPL) or from mesenteric lymph nodes (MnL), whereas IgA of plasmacytoma origin against S . tel aviv and purified intestinal sIgA against Shigella X16 induced specific antibacterial ADCC with both SpL and PPL . When sIgA were tested with intestinal lymphocytes from the epithelium and the lamina propria, i.e., cells from the gut mucosa which first interact with enteric bacteria, it was found that both these lymphoid populations were able to express sIgA-dependent ADCC against Shigella X16 . In parallel tests, cells from thymus and popliteal lymph nodes failed to express ADCC . Blocking studies with purified IgG and IgA of goat, rabbit, and mouse origin demonstrated that the Fc-alpha and Fc-gamma receptors were specifically involved in IgA- or IgG-dependent antibacterial ADCC . At least two effector populations, a macrophage and a Thy-1.2- lymphocyte, were observed to exert IgA-ADCC at the splenic level, whereas only lymphoid cells expressed this activity at the GALT level . Together, these results describe a new activity of IgA against enteropathogenic bacteria. Mutat Res, 1984 Aug, 140(4), 169 - 74 Structure-dependent variation in the mutagenic, prophage-inducing and antibacterial activities of 5-nitro-2-furamide derivatives; Kato Y et al.; A comparative survey of the mutagenic, prophage-inducing and antibacterial activities of 3 structure-related series of 5-nitro-furan derivatives including 5-nitro-2-furohydrazide imide, 5-nitro-2-furamide oxime and 5-nitro-2-furohydrazide has been undertaken . Among the compounds assayed, the 5-nitro-2-furohydrazide imide series was found to be most active with regard to mutagenic and antibacterial activities against Salmonella typhimurium TA100 and prophage-inducing activity in Escherichia coli GY5027 . A clear correlation was observed between the chemical structure and the mutagenic and prophage-inducing activities which were approximately correlated to the antibacterial activity. J Immunol, 1984 Aug, 133(2), 950 - 7 Monoclonal antibodies to Salmonella lipopolysaccharide: anti-O-polysaccharide antibodies protect C3H mice against challenge with virulent Salmonella typhimurium; Colwell DE et al.; The present investigation reports the production of monoclonal antibodies to antigenic determinants of the O-polysaccharide of Salmonella typhimurium lipopolysaccharide (LPS), and assesses the effectiveness of these antibodies in protecting C3H mice against the lethal effects of Salmonella infection . Hybridomas were generated by fusing spleen cells from (BALB/c X A/J)F1 (CAF1) mice hyperimmunized by i.v . injection with acetone-killed S . typhimurium SR-11 with X63-Ag8.653 murine myeloma cells . Hybridomas producing antibodies reactive with S . typhimurium SR-11 whole cells were subcloned, and seven of the resulting clones as well as one previously described clone were selected for use in the studies reported here . Monoclonal antibodies from these eight clones were of the IgG1 (1), IgG3 (6), or IgM (1) isotype and were specific for the O-polysaccharide region of Salmonella LPS, reacting with LPS from smooth S . typhimurium SR-11 and LT-2, but not with LPS from rough S . minnesota R60 (Ra), R345 (Rb), or R595 (Re) . The effectiveness of each monoclonal antibody in protecting C3H/HeN and C3H/HeJ mice against the lethal effects of Salmonella infection was evaluated by comparing the median length of survival of groups of mice given antibody by i.p . injection before i.p . challenge with virulent S . typhimurium SR-11 to that of animals that received no antibody . Three out of eight monoclonal anti-O-polysaccharide antibodies, ST-1 (IgM), 10-5-47 (IgG3), and 10-5-6 (IgG3), provided significant (p less than 0.01) protection to C3H/HeN mice challenged with approximately 10(4) LD100 of Salmonella . Only antibodies ST-1 and 10-5-6, however, extended the median length of survival of C3H/HeJ mice beyond that of infected controls . Mouse antiserum prepared against S . typhimurium SR-11 was equally protective in C3H/HeJ mice . In an attempt to understand the contribution of antibody specificity to the relative differences in the protective capacities of the monoclonal antibodies, their reactivities with several Salmonella reference strains of different classical serotypes were examined . Although some differences in reactivity against the different strains were apparent, this approach was not adequate for defining the fine specificity of these monoclonal antibodies . The results of this study provide evidence that monoclonal antibodies with specificity to the O-polysaccharide region of Salmonella LPS can protect C3H mice against challenge with the homologous bacterial strain. Can J Microbiol, 1984 Aug, 30(8), 991 - 6 Evidence for involvement of pyrH+ of an Escherichia coli K-12 F-prime factor in inhibiting construction of hybrid merodiploids with Salmonella typhimurium; Kelln RA; Transconjugants were not recovered in matings between Salmonella typhimurium and Escherichia coli K-12 strains carrying the chromosomal region between leu and argF on an F-prime factor, even when a restriction-deficient recipient was used . A mutant F-prime factor compatible for transfer to S . typhimurium was constructed by transposon mutagenesis and characterized as being deficient in directing the synthesis of UMP kinase (encoded by pyrH) . Other compatible F-prime factors were readily constructed by employing a procedure designed to select for strains carrying F-prime factors harboring pyrH mutations. EMBO J, 1984 Aug, 3(8), 1745 - 52 The genetic control of DNA supercoiling in Salmonella typhimurium; Richardson SM et al.; We have elucidated the genetic control of DNA supercoiling in Salmonella typhimurium . The level of superhelix density is controlled by two classes of genes . The only member of the first class is topA, the structural gene for topoisomerase I . The second class, tos, (topoisomerase one suppressor) consists of at least two genes, one of which is linked to gyrA, the structural gene for the topoisomerase subunit of DNA gyrase . Deletions of topA result in oversupercoiling of plasmid DNA . These mutations do not require the acquisition of second-site compensatory mutations to allow cell growth, in contrast to the situation in Escherichia coli . However, tos mutations, unlinked to topA, have been isolated which reduce plasmid superhelix density . We conclude that the level of DNA supercoiling in S . typhimurium is a dynamic balance between the effects of the gene products of topA (relaxation) and tos (supercoiling) which act independently of each other . Using a variety of combinations of these mutations we have constructed a series of isogenic strains, each of which has a different but precisely defined level of plasmid supercoiling; the series as a whole provides a wide range of supercoiling both above and below the wild-type level. Pediatr Res, 1984 Aug, 18(8), 789 - 94 Neutrophil bactericidal dysfunction towards oxidant radical-sensitive microorganisms during experimental iron deficiency; Moore LL et al.; We developed a clear-cut nutritional iron deficiency anemia without concomittant malnutrition in rats given a low iron diet, and we restored normal iron and hemoglobin levels in these same animals with iron dextran injections . The neutrophil function studies performed during and after a period of iron deficiency showed the following: Phagocytosis of Staphylococcus aureus 502A, Streptococcus pneumoniae, and Salmonella typhimurium was not altered by iron deficiency or by the administration of iron; phagocytosis of Candida albicans was moderately abnormal during iron deficiency, and became normal with the restoration of iron sufficiency . Microbicidal activity towards Staphylococcus aureus 502A and Candida albicans, two catalase-positive microorganisms, was markedly decreased (to 50% of control values) and returned to normal when iron sufficiency was restored . Killing of a catalase-negative organism, Streptococcus pneumoniae was normal in iron-deficient rats . This pattern of differential bactericidal activities suggested an abnormality of the oxidant radical-generating machinery in neutrophils of iron-deficient animals . Indeed, iron deficiency caused a marked decrease of neutrophil nitroblue tetrazolium dye reduction, which disappeared after iron administration . Neutrophil myeloperoxidase activity was slightly decreased in iron deficient rats and returned to normal after iron administration . Microbicidal activity towards a gram-negative, catalase-positive organism, Salmonella typhimurium, was equal in iron deficient and iron sufficient animals . Our combined results suggest that a definite microbicidal defect is the consequence of nutritional iron deficiency, apart from any protein-calorie malnutrition . This defect affects the disposal in PMNs of two catalase-positive microorganisms (which require intracellular production of oxidant radicals for their destruction) but not of a catalase-negative bacterial species.(ABSTRACT TRUNCATED AT 250 WORDS) J Bacteriol, 1984 Aug, 159(2), 618 - 23 Location and analysis of nucleotide sequences at one end of a putative lac transposon in the Escherichia coli chromosome; Buvinger WE et al.; A segment of Escherichia coli DNA that contained a discontinuity of homology with Salmonella typhimurium DNA was isolated . The segment, 1,430 base pairs long, was derived from one end of the lac "loop," a region of about 12 kilobase pairs of E . coli DNA, including the lac operon which has no detectable homology with S . typhimurium DNA (K . Lampel and M . Riley, Mol . Gen . Genet . 186:82-86, 1982) . The nucleotide sequence of the 1,430-base-pair segment of DNA was determined . The location of the junction of discontinuity of homology within the segment was established by hybridization experiments . Nucleotide sequences at or near the junction were determined to be similar to sequences that are involved in site-specific inversion in S . typhimurium, E . coli, phage P1, and phage Mu . Similar sequences are also present within the terminal inverted repeat sequences of transposon Tn5 and at the V-D-J joining sequences of eucaryotic immunoglobulin genes . Therefore, the lac operon, together with flanking DNA, may have been inserted into the E . coli chromosome at one time via a site-specific recombination event . Rearrangement events of this kind undoubtedly have played a significant role in the evolutionary divergence of chromosomal DNAs. J Bacteriol, 1984 Aug, 159(2), 453 - 9 Aspartate-specific peptidases in Salmonella typhimurium: mutants deficient in peptidase E; Carter TH et al.; The only dipeptide found to serve as a leucine source for a Salmonella strain lacking peptidases N, A, B, D, P, and Q was alpha-L-aspartyl-L-leucine . A peptidase (peptidase E) that specifically hydrolyzes Asp-X peptides was identified and partially purified from cell extracts . The enzyme (molecular weight, 35,000) is inactive toward dipeptides with N-terminal asparagine or glutamic acid . Mutants (pepE) lacking this enzyme were isolated by screening extracts for loss of the activity . Genetic mapping placed the pepE locus at 91.5 map units and established the gene order metA pepE zja-861::Tn5 malB . Duplications of the pepE locus showed a gene dosage effect on levels of peptidase E, suggesting that pepE is the structural gene for this enzyme . Mutations in pepE resulted in the loss of the ability to grow on Asp-Pro as a proline source but did not affect utilization of other dipeptides with N-terminal aspartic acid . Loss of peptidase E did not cause a detectable impairment in protein degradation . Two other peptidases present in cell extracts of mutants lacking peptidases N, A, B, D, P, Q, and E also hydrolyze many Asp-X dipeptides. J Biol Chem, 1984 Jul 25, 259(14), 8753 - 7 The sulfonylurea herbicide sulfometuron methyl is an extremely potent and selective inhibitor of acetolactate synthase in Salmonella typhimurium; LaRossa RA et al.; The sulfonylurea herbicide sulfometuron methyl inhibits the growth of several bacterial species . In the presence of L-valine, sulfometuron methyl inhibits Salmonella typhimurium, this inhibition can be reversed by L-isoleucine . Reversal of growth retardation by L-isoleucine, accumulation of guanosine 5'-diphosphate 3'-diphosphate (magic spot), and relA mutant hypersensitivity suggest sulfometuron methyl interference with branched-chain amino acid biosynthesis . Growth inhibition of S . typhimurium is mediated by sulfometuron methyl's inhibition of acetolactate synthase, the first common enzyme in the branched-chain amino acid biosynthetic pathway . Sulfometuron methyl exhibits slow-binding inhibition of acetolactate synthase isozyme II from S . typhimurium with an initial Ki of 660 +/- 60 nM and a final, steady-state Ki of 65 +/- 25 nM . Inhibition of acetolactate synthase by sulfometuron methyl is substantially more rapid (10 times) in the presence of pyruvate with a maximal first-order rate constant for conversion from initial to final steady-state inhibition of 0.25 +/- 0.07 min-1 (minimal half-time of 2.8 min) . Mutants of S . typhimurium able to grow in the presence of sulfometuron methyl were obtained . They have acetolactate synthase activity that is insensitive to sulfometuron methyl because of mutations in or near ilvG, the structural gene for acetolactate synthase isozyme II. J Mol Biol, 1984 Jul 25, 177(1), 1 - 18 A repetitive DNA sequence, rhs, responsible for duplications within the Escherichia coli K-12 chromosome; Lin RJ et al.; A novel family of large, imperfectly repeated DNA sequences has been found in Escherichia coli . Two members of this family, rhsA and rhsB, occur as direct repeats, flanking the pit glyS xyl segment of the chromosome . Unequal sister-chromatid crossing over between rhsA and rhsB accounts for the frequent tandem duplication of the glyS locus that has been observed by various workers . This unequal recombination is recA-dependent . The rhsA locus is operationally defined as the segment between xyl and mtl that is repeated at other chromosomal locations . Using this definition, rhsA extends minimally 5500 base-pairs; 3800 base-pairs of rhsA are sufficiently homologous to rhsB to form an S1 nuclease-resistant heteroduplex with it . The rhsA sequence also exhibits internal repetition . At least one additional rhs sequence occurs in the E . coli chromosome unlinked to either rhsA or rhsB . Southern analysis of restriction digests of genomic DNA from E . coli strains C and B/5 showed that both of these strains have rhs hybridizable patterns similar to strain K-12, but the rhs sequence is absent in Salmonella typhimurium . The function of the rhs sequences has not been discovered . In the course of this work we developed a technique, termed "transductional walking", by which chromosomal DNA adjacent to a previously cloned DNA segment can be cloned through genetic procedures. Carcinogenesis, 1984 Jul, 5(7), 853 - 6 Mutagenicity of gastric juice: the importance of controlling histidine concentration when using Salmonella tester strains; O'Connor HJ et al.; The mutagenic activity of gastric juice has been assessed using bacterial tester strains that undergo reverse mutation (Salmonella typhimurium:his(-)----his(+)) . Free histidine, a known source of inaccuracy in this mutation test system, was detected in 42 of 73 juice samples (concentration range 3.5-992.4 micrograms/ml); high histidine concentrations were significantly correlated with hypochlorhydria . The effect of histidine was controlled by using a pre-incubation modification of the Salmonella fluctuation test in which juice samples and their corresponding control cultures containing equivalent amounts of histidine were incubated with the tester bacteria prior to plating out . Significant mutagenic activity was found in a high proportion of samples (18 of 20) . The histidine content in gastric juice which can affect in vitro mutagenicity testing must be adequately controlled before positive or negative results can be equated with the presence or absence of intragastric carcinogens. Xenobiotica, 1984 Jul, 14(7), 545 - 8 Glutathione transferase-mediated and non-enzymatic activation and detoxication of the N-hydroxy derivative of Trp-P-2, a potent pyrolysate promutagen; Saito K et al.; Glutathione (GSH) transferase-mediated and non-enzymatic activation and detoxication of 3-hydroxyamino-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-b}indole (Trp-P-2), a mutagenic and carcinogenic heterocyclic amine . The enzymatic GSH conjugation with N-OH-Trp-P-2 was catalysed by rat-liver GSH transferase and a rat-liver cytosol fraction to form three conjugates (CH-1, CH-2 and CH-3) . The mutagenicities of the GSH conjugates 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, but not to N-OH-Trp-P-2 formed from it. Rev Infect Dis, 1984 Jul-Aug, 6(4), 439 - 43 Separation and characterization of toxic and nontoxic forms of lipid A; Takayama K et al.; Highly purified and well-characterized samples of precursors and derivatives of bacterial lipopolysaccharides (LPS) were used to study the relationship between the chemical structure of lipid A and its toxicity . These samples included lipids X and Y (monosaccharide precursors of lipid A) from Escherchia coli MN7; incomplete lipid A (a disaccharide precursor of lipid A) from Salmonella typhimurium i50; and monophosphoryl lipid A, TLC-3 (a derivative of LPS), from S . typhimurium G30/C21 . In addition, a diphosphoryl lipid A, TLC-3, was prepared from LPS of S . typhimurium G30/C21 and characterized by positive fast atom bombardment mass spectrometry . The diphosphoryl lipid A, TLC-3 fraction, was determined to be very toxic by the chick embryo lethal-dose test (CELD50 = 0.0064 microgram) . Lipids X and Y, incomplete lipid A, and monophosphoryl lipid A (TLC-3), were all nontoxic (CELD50 greater than 10 micrograms) . These results suggest a multiple structural requirement for toxicity of lipid A . Toxic lipid A must contain all of the following components: a glucosamine disaccharide, a sugar-1-phosphate, and normal fatty acid(s). Mutat Res, 1984 Jul, 137(1), 17 - 28 Genotoxicity of apomorphine and various catecholamines in the Salmonella mutagenicity test (Ames test) and in tests for primary DNA damage using DNA repair-deficient B . subtilis strains (rec assay); Suter W et al.; Apomorphine, N-nor-N-propyl-apomorphine, dopamine, L-DOPA, 6-hydroxydopamine and adrenaline were evaluated for genotoxicity using the Ames test and DNA repair-deficient and DNA repair-proficient Bacillus subtilis strains (rec assay, H17/M45; HLL3g/HJ-15) . In the absence of an S9 liver homogenate, apomorphine induced frame-shift mutations in Salmonella typhimurium, mainly in strain TA1537; no indication of DNA-damaging effects in B . subtilis was observed . N-Nor-N-propyl-apomorphine was tested using strain TA1537 only and found to be mutagenic . Dopamine, L-DOPA, 6-hydroxydopamine and adrenaline were non-mutagenic when tested without S9, whereas they were all more toxic for DNA repair-deficient than for DNA repair-proficient B . subtilis strains, indicating a DNA-damaging potential . In a second set of experiments the mode of action of apomorphine and the relevance of the positive Ames test data were investigated . Glutathione in physiological concentrations reduced the mutagenic effect of apomorphine in a dose-dependent way, both in the presence and the absence of S9 . S9 also reduced the mutagenicity of apomorphine . By comparing the effects of a complete S9 mix with those of a preparation without glucose-6-phosphate and NADP, it became clear that S9 also had an activating effect, overshadowed under standard conditions by its deactivating activity . Apomorphine was not mutagenic under anaerobic conditions . Superoxide dismutase and catalase reduced the mutagenic effect of apomorphine . All test conditions which reduced the mutagenic effect also inhibited the dark discoloration of the tester plates, indicating a retardation of apomorphine oxidation . It can, therefore, be concluded that oxidation of apomorphine leads to mutagenic products which induce frame-shift mutations in Salmonella typhimurium . This oxidation was prevented both by glutathione in concentrations well below physiological levels and/or by catalase and superoxide dismutase . Under these conditions, apomorphine was non-mutagenic in therapeutic concentrations as well as at higher dose levels . The possibility of genotoxic side effects occurring in patients treated with apomorphine as an emetic drug is therefore considered to be very unlikely. Mutat Res, 1984 Jul, 127(2), 113 - 8 Effect of Japanese seaweed (Laminaria angustata) extracts on the mutagenicity of 7,12-dimethylbenz{a}anthracene, a breast carcinogen, and of 3,2'-dimethyl-4-aminobiphenyl, a colon and breast carcinogen; Reddy BS et al.; Animal model studies suggest that diets containing Laminaria angustata, a brown seaweed commonly eaten in Japan, inhibit breast carcinogenesis . In order to identify the compound(s) in the seaweed responsible for tumor-inhibiting activity, we used Ames/mammalian microsome assay system to determine the antimutagenic (or anticarcinogenic) effect of various solvents and water extracts of Laminaria angustata . The antimutagenic effects of acetone, ether, chloroform, chloroform + methanol, hot water and cold water extracts on the mutagenicity induced by 7,12-dimethylbenz{a}anthracene (DMBA), a breast carcinogen, and 3,2'-dimethyl-4-aminobiphenyl (DMAB), a colon and breast carcinogen, was studied using the Salmonella typhimurium strains TA98 and TA100 . All extracts were nonmutagenic in both bacterial tester strains . The addition of 10-100 mg solvent extracts of seaweed/plate greatly inhibited DMAB-induced mutagenicity in both tester strains (80-96% inhibition) and DMBA-induced mutagenicity in TA100 (about 82%), whereas hot and cold water extracts produced a moderate inhibition in a dose-related manner in both strains. Food Chem Toxicol, 1984 Jul, 22(7), 535 - 9 Inactivation of quercetin mutagenicity; Friedman M et al.; Combinations of oxygen and alkaline pH were found to inactivate irreversibly the mutagenicity of quercetin towards Salmonella typhimurium strain TA98 . Exposure time, quercetin concentration and polyphenol oxidase were also important variables determining the extent of quercetin inactivation . Temperature had a relatively weak influence on the extent of inactivation . The metal salts, ferrous, ferric and copper sulphates also brought about inactivation but this effect was partially reversed when the pH of the incubation medium was reduced from 7 to 2 . Ferric sulphate had a much smaller effect than did the ferrous salt except in the presence of tyrosinase and oxygen at pH 7 . Zinc sulphate impaired quercetin mutagenicity only very slightly in the presence of tyrosinase and oxygen . When an oxygen-saturated solution of quercetin was exposed to tyrosinase at various pH values, the ultraviolet absorption maximum of quercetin near 370 nm decreased to an extent that correlated with the mutagenicity of quercetin under those conditions. Can J Microbiol, 1984 Jul, 30(7), 916 - 21 Tricarboxylate transport in a Cit+ Escherichia coli: evidence for the role of an outer membrane protein; Tomas JM et al.; A strain of Escherichia coli of bovine origin able to use tricarboxylates as single carbon source is described . Tricarboxylate utilization (Cit+) and fluorocitrate sensitivity (FCs) could be transferred conjugatively to E . coli K12 and were not plasmid borne . No evidence was found for tct gene products of Salmonella typhimurium . A citrate-inducible outer membrane protein of 21-22 kilodaltons (kd) was found only in Cit+ strains . A protein (21-22kd) protein was also found in wild-type E . coli K12 and in fluorocitrate-resistant mutants of Cit+ strains, but it was present in a cryptic form no longer inducible by citrate . Fluorocitrate-resistant mutants of Cit+ strains were still able to transport citrate by a fluorocitrate-insensitive system . High levels of the 22-kd protein correlated with reduced growth induction times on citrate and with the ability to effectively transport citrate. Rev Infect Dis, 1984 Jul-Aug, 6(4), 567 - 72 Lipid A and immunotherapy; Ribi E et al.; Endotoxin isolated from Re mutants of Salmonella typhimurium or Salmonella minnesota and consisting only of 3-deoxy-D-mannooctulosonic acid (KDO) and lipid A synergistically enhances the ability of mycobacterial cell wall skeleton (CWS) to regress transplantable, line-10 tumor (hepatocellular carcinoma) in syngeneic guinea pigs . Tumor regression is rapid, and systemic tumor immunity concomitantly develops when as little as 50 micrograms of each of these two components is combined and injected intralesionally . Selective removal of KDO from endotoxin yields diphosphoryl lipid A, which retains its toxic properties . Subsequent selective removal of the phosphate moiety at the reducing end of the diphosphoryl lipid A molecule yields nontoxic, monophosphoryl lipid A (determined by lethality for chick embryos) . Like the parent endotoxin or toxic diphosphoryl lipid A, monophosphoryl lipid A retains the ability to synergistically enhance the antitumor activity of mycobacterial CWS adjuvant . Both di- and monophosphoryl lipid A contain mixtures of a series of structural analogs . They can be separated chromatographically into single components that differ in number, type, and position of ester-linked fatty acids . Comparison of chromatographic fractions reveals that components of toxic and nontoxic lipid A can be paired according to structure . Each component of the pair has the same molecular structure, with the exception of an additional phosphate group in the toxic component . The toxicity of "lipid A's" liberated from endotoxin by acid hydrolysis appears to be determined by the proportion of di- and monophosphoryl lipid A in the hydrolysis mixture . Structural analogs of monophosphoryl lipid A, which differ in degree of O-acylation and type and distribution of fatty acids, have comparable antitumor activity.(ABSTRACT TRUNCATED AT 250 WORDS) Rev Infect Dis, 1984 Jul-Aug, 6(4), 535 - 41 Stimulation of spleen cells and macrophages of C3H/HeJ mice by a lipid A precursor derived from Salmonella typhimurium; Vogel SN et al.; Lipid A is that portion of the lipopolysaccharide (LPS) molecule believed to mediate most of the biologic activities associated with protein-free endotoxic preparations . C3H/HeJ mice possess a mutation at the Lps gene locus (Lpsd) that results in a state of profound hyporesponsiveness to the biologic effects of LPS (and more specifically, lipid A) in vivo . The relative unresponsiveness in vivo to LPS exhibited by these mice is reflected at a cellular level, as evidenced by a failure of many different cell types derived from the C3H/HeJ strain to respond to LPS or lipid A in vitro; this lack of response contrasts with that of cell cultures prepared from endotoxin responsive (Lpsn) mouse strains . Evidence is presented which demonstrates that a lipid A precursor molecule, produced by a mutant of Salmonella typhimurium conditionally defective in the synthesis of 3-deoxy-D-mannooctulosonic acid, stimulates mitogenesis in C3H/HeJ splenic cultures and induces cultures of C3H/HeJ macrophages to produce significant levels of the monokine interleukin-1 (IL-1; previously referred to as lymphocyte activating factor or LAF) and prostaglandins of the E series . These findings suggest the possibility that the failure of C3H/HeJ cells to respond to intact LPS or lipid A may be related to a defect in the processing of lipid A or LPS to a suitably stimulatory form, rather than to a defect in the recognition of the lipid A region. Rev Infect Dis, 1984 Jul-Aug, 6(4), 455 - 62 Effect of altered lipid A synthesis on the synthesis of major proteins of the Salmonella typhimurium outer membrane; Rick PD et al.; The effect of altered lipid A synthesis on the synthesis of major outer membrane proteins was investigated in mutants of Salmonella typhimurium conditionally defective in the synthesis of 3-deoxy-D-mannooctulosonic acid (KDO) . The defect is due to a mutation in the structural gene for KDO-8-phosphate synthetase (designated kdsA), and expression of this lesion results in the accumulation of a precursor of lipid A that not only lacks KDO but is also deficient in ester-linked fatty acyl residues . During the initial 20-30 min following a shift of mutants to nonpermissive conditions, the rate of synthesis of the OmpA protein increased approximately 2.5-fold and then decreased . In contrast, the rates of synthesis of total cell-envelope proteins, as well as that of the porin proteins, were unaffected . The mechanism responsible for the initial increase in the rate of OmpA synthesis remains to be established . However, it appears that the subsequent decrease in the rate of OmpA synthesis may be related to a decrease in the stability of OmpA messenger RNA . The effect of nonpermissive conditions on the rate of OmpA synthesis was specifically related to expression of the kdsA lesion, and it was not found to be strain-specific or uniquely related to a single kdsA mutant allele. Atherosclerosis, 1984 Jul, 52(1), 123 - 6 Effect of bacterial lipopolysaccharide on serum high density lipoprotein cholesterol in rabbits; Kerttula Y et al.; The effect of bacterial lipopolysaccharide (LPS) on serum lipids was examined in rabbits . LPS was prepared from the smooth Salmonella typhimurium LT2 strain and given intravenously at a dose of 100 ng/kg b.wt . There were no significant changes in serum triglyceride or cholesterol levels in 1-3 days after the administration of LPS . There was, however, a decrease in serum high density lipoprotein (HDL) cholesterol, which was greatest after 2 days (P less than 0.001) . Simultaneously, the HDL/total cholesterol ratio decreased (P less than 0.005). Toxicol Lett, 1984 Jul, 22(1), 15 - 20 Evaluation of the mutagenic potential of endod (Phytolacca dodecandra), a molluscicide of potential value for the control of schistosomiasis; Pezzuto JM et al.; Extracts of the fruit of Phytolacca dodecandra (endod) demonstrate molluscicidal and other biological activities . Since this plant is indigenous to some countries where schistosomiasis is a common problem, it has been proposed that it may be socioeconomically feasible to employ endod as an aid in the control of this disease through its use to control the snail vector . As an initial step in the safety assessment of this substance, its mutagenic potential was determined utilizing Salmonella typhimurium strain TM677 . The seeds and fruit of Phytolacca americana, also molluscicidal, were additionally evaluated for mutagenic potential . Using a variety of conditions, no mutagenic activity could be demonstrated for any of the extracts tested . Thus, subject to the results of future safety assessment, endod remains a viable candidate as a useful molluscicide. Mutat Res, 1984 Jul, 137(1), 7 - 15 Lack of mutagenicity of synthetic pyrethroids in Salmonella typhimurium strains and in V79 Chinese hamster cells; Pluijmen M et al.; Seven pyrethroids, i.e., cypermethrin, permethrin, deltamethrin, bioresmethrin, resmethrin, cismethrin and fenvalerate, were not found to be mutagenic in (a) Salmonella typhimurium strains TA100 or TA98 in the presence or absence of a rat liver activation system using the plate incorporation assay and fluctuation tests, or (b) V79 Chinese hamster cells in the presence or absence of hepatocytes. Mutat Res, 1984 Jul, 137(1), 39 - 45 The effect of acetyl-CoA supplementation on the mutagenicity of benzidines in the Ames assay; Kennelly JC et al.; The conventional Ames assay metabolising system was confirmed to be deficient in its ability to N-acetylate . This may render the test less sensitive to compounds which normally have an acetylation step during their in vivo activation to carcinogens . The addition of acetyl-coenzyme A to the S9 mix in the Ames assay increased the mutagenicity of benzidine in Salmonella typhimurium strains TA98 and TA1538 4-5-fold . This was consistent with the observation that benzidine is N-acetylated prior to DNA binding in vivo in rat liver . Two 3,3'-disubstituted benzidines, o-tolidine and o-dianisidine, were also tested . A smaller increase in o-tolidine mutagenicity, compared to that observed with benzidine, occurred with the addition of acetyl-coenzyme A . However, the production of acetylated metabolites from o-tolidine was only 37% of that from benzidine . The mutagenicity of o-dianisidine was unaffected by acetyl-coenzyme A . Acetylation of o-dianisidine was only 16% of that observed with benzidine, and the N-acetyl derivatives of o-dianisidine showed lower mutagenicity than the parent amine . The differing responses of benzidine, o-tolidine and o-dianisidine to addition of acetyl-coenzyme A suggests it may not be possible to simply infer the metabolism of 3,3'-disubstituted benzidines to DNA binding species from data on benzidine itself. Mutat Res, 1984 Jul, 137(1), 29 - 32 Mutagenicity of some synthetic quinolines and quinoxalines related to IQ, MeIQ or MeIQx in Ames test; Grivas S et al.; 3-Methyl- and 3,4-dimethyl-3H-imidazo{4,5-f}quinoline, 3,8-dimethyl-3H-imidazo{4,5-f}quinoxaline, N6-methyl- and N6,7-dimethylquinoline-5,6-diamine, as well as N6,3-dimethylquinoxaline-5,6-diamine, have been synthesized . Only the first-mentioned compound was active in Ames test; the response was equal for Salmonella typhimurium TA98 and TA100, regardless of enzymatic activation (S9) . However, its mutagenicity to TA98 + S9 was 300-1300 times smaller than the values reported for the related compounds, 3-methyl- and 3,4-dimethyl-3H-imidazo{4,5-f}quinolin-2-amine ('IQ' and 'MeIQ'), and for 3,8-dimethyl-3H-imidazo{4,5-f}quinoxalin-2-amine ('MeIQx') . Hence, the presence of the imidazole ring and the 2-amino group in the molecule seems to be important for the high mutagenicity of the latter compounds. Br J Cancer, 1984 Jul, 50(1), 91 - 6 Mutagenic and cytotoxic activity of doxorubicin and daunorubicin derivatives on prokaryotic and eukaryotic cells; Babudri N et al.; The mutagenic and cytotoxic activity of two newly synthesized doxorubicin derivatives and of one daunorubicin derivative were studied in V79 Chinese hamster cells and bacteria (Salmonella typhimurium and Escherichia coli) . The results showed that all the compounds tested were cytotoxic and mutagenic for both prokaryotic and eukaryotic cells . However, in both systems, the two 4-desmethoxy- and the 4'-desoxy-derivatives were more active than the parent compounds, indicating that modifications in the aglycone or in the sugar moiety can produce appreciable changes in the biological properties of the anthracycline antibiotics . The in vitro activities observed in this study correlated with the in vivo antitumour potency. J Bacteriol, 1984 Jul, 159(1), 206 - 13 Salmonella typhimurium synthesizes cobalamin (vitamin B12) de novo under anaerobic growth conditions; Jeter RM et al.; In this paper, we report that the enteric bacterium Salmonella typhimurium synthesized cobalamin de novo under anaerobic culture conditions . Aerobically, metE mutants of S . typhimurium need either methionine or cobalamin as a nutritional supplement for growth . The growth response to cobalamin depends upon a cobalamin-requiring enzyme, encoded by the gene metH, that catalyzes the same reaction as the metE enzyme . Anaerobically, metE mutants grew without any nutritional supplements; the metH enzyme functioned under these conditions due to the endogenous biosynthesis of cobalamin . This conclusion was confirmed by using a radiochemical assay to measure cobalamin production . Insertion mutants defective in cobalamin biosynthesis (designated cob) were isolated in the three major branches of the cobalamin biosynthetic pathway . Type I mutations blocked the synthesis of cobinamide, type II mutations blocked the synthesis of 5,6-dimethylbenzimidazole, and type III mutations blocked the synthesis of cobalamin from cobinamide and 5,6-dimethylbanzimidazole . Mutants that did not synthesize siroheme (cysG) were blocked in cobalamin synthesis . Genetic mapping experiments showed that the cob mutations are clustered in the region of the S . typhimurium chromosome between supD (40 map units) and his (42 map units) . The discovery that S . typhimurium synthesizes cobalamin de novo only under anaerobic conditions raises the possibility that anaerobically grown cells possess a variety of enzymes which are dependent upon cobalamin as a cofactor. Infect Immun, 1984 Jul, 45(1), 62 - 6 Mouse fibroblast interferon modifies Salmonella typhimurium infection in infant mice; Bukholm G et al.; The effect of mouse fibroblast interferon on Salmonella typhimurium infection in infant mice was examined . The lethality to mice that had been given S . typhimurium intragastrically was significantly reduced in a dose-dependent manner when the mice were pretreated with fibroblast interferon . Lower doses of interferon delayed the development of disease . Interferon neutralized with anti-interferon globulin did not influence the lethality of S . typhimurium to mice . In mice treated with interferon there was also a reduced invasiveness of S . typhimurium in intestinal epithelial cells in vivo . It was further demonstrated in an in vitro system that interferon pretreatment of mouse L-929 cells inhibited the invasiveness of the bacteria in a dose-dependent manner . The in vitro inhibition was neutralized with anti-interferon globulin . The results indicate that interferon inhibits Salmonella bacteria from invading cells and establishing an intracellular state of infection . This may represent an important factor in the pathogenesis of disease. Infect Immun, 1984 Jul, 45(1), 29 - 35 Cationic antimicrobial proteins isolated from human neutrophil granulocytes in the presence of diisopropyl fluorophosphate; Shafer WM et al.; Acid (0.2 M sodium acetate, pH 4.0) extracts of granules recovered from disrupted human polymorphonuclear granulocytes (PMNs) exhibited in vitro antimicrobial activity against Salmonella typhimurium . To minimize proteolytic destruction or modification of antimicrobial proteins derived from these granules, we pretreated the PMNs with the serine protease inhibitor diisopropyl fluorophosphate . Fractionation of such extracts by carboxymethyl Sephadex and Sephadex G-75 chromatography resulted in the recovery of at least two antimicrobial, cationic proteins . These proteins differed substantially in antimicrobial activity, amino acid composition, and molecular weight (Mr, 37,000 and 57,000) . As we have shown before (Shafer et al., Infect . Immun . 43:834-858), with unfractionated proteins, these two proteins exhibited diminished activity against a polymyxin B-resistant (PBr) mutant of S . typhimurium compared with their activity against the isogenic parental polymyxin B-sensitive (PBs) strain . Expression of the relevant mutation (prmA) in the PBr mutant decreases the electronegativity of lipid A, owing to increased 4-amino-4-deoxy-L-arabinosylation at the 4' phosphate residue (Vaara et al., FEBS Lett . 129:145-149) . The data suggest that at least two different cationic proteins account for the antimicrobial capacity of extracts from human PMN granules . Moreover, the availability of anionic charges in the outer membrane of S . typhimurium due to free lipid A phosphates apparently dictates phenotypic levels of resistance to both of the cationic proteins extracted from human PMN granules. Cancer Res, 1984 Jul, 44(7), 2848 - 54 Molecular requirements for the mutagenicity of malondialdehyde and related acroleins; Basu AK et al.; Malondialdehyde, a product of lipid peroxidation and prostaglandin biosynthesis, is mutagenic in Salmonella . To determine the molecular requirements for its mutagenicity, we tested a series of beta-substituted acroleins in Salmonella typhimurium hisD3052 . Mutagenicity is dependent on the steric bulk of the substituent (revertants/mumol) at the beta position: beta- methoxyacrolein , 220; beta- ethoxyacrolein , 110; and beta- isobutoxyacrolein , 40 . A good leaving group at the beta position substantially increases the mutagenic activity (revertants/mumol): beta-(p-nitrophenoxy)acrolein, 620; beta- benzoyloxyacrolein , 320; beta- chloroacrolein , 890; and di-gamma- oxopropenyl ether, 870 . These data suggest that nucleophilic attack on the beta-carbon followed by elimination of the beta substituent is important for mutagenicity . Substitution of a methyl group at the alpha-carbon abolishes mutagenicity of these compounds . This effect can be explained by the lack of chemical reactivity of the alpha-methyl analogues toward oxygen or nitrogen nucleophiles . Propynal , which can add nucleophiles to generate a substituted acrolein, exhibits the highest mutagenicity (1370 revertants/mumol) in this series . The importance of the aldehyde functionality is suggested by the nonmutagenicity of propiolonitrile , ethyl propiolate , 4-benzoyloxy-3- buten -2-one, and 4-methoxy-3- buten -2-one . Aldehyde addition subsequent to the formation of the Michael adduct is, therefore, important for mutagenesis . An investigation of the toxicity of the present series indicates that toxicity and mutagenicity are independent events based on different chemical reactions. J Bacteriol, 1984 Jul, 159(1), 130 - 7 Conditionally transposition-defective derivative of Mu d1(Amp Lac); Hughes KT et al.; A Mu d1 derivative is described which is useful for genetic manipulation of Mu-lac fusion insertions . A double mutant of the specialized transducing phage Mu d1(Amp Lac c62ts) was isolated which is conditionally defective in transposition ability . The Mu d1 derivative, designated Mu d1-8(Tpn{Am} Amp Lac c62ts), carries mutations which virtually eliminate transposition in strains lacking an amber suppressor . In such strains, the Mu d1-8 prophage behaves like a standard transposon . It can be moved from one strain of Salmonella typhimurium to another by the general transducing phage P22 with almost 100% inheritance of the donor insertion mutation . When introduced into a recipient carrying supD, supE, or supF, 89 to 94% of the Ampr transductants were transpositions of the donor Mu d1-8, from the transduced fragment into new sites . The stability of Mu d1-8 in a wild-type, suppressor-free background was sufficient to permit use of the fusion to select constitutive mutations without prior isolation of deletions to stabilize the fusion . Fusion strains could be grown at elevated temperature without induction of the Mu d prophage . The transposition defect of Mu d1-8 was corrected by a plasmid carrying the Mu A and B genes. Acta Pathol Microbiol Immunol Scand {A}, 1984 Jul, 92(4), 195 - 204 The effect of exercise and fasting on the myocardial protein and lipid metabolism in experimental bacterial myocarditis; Ilback NG et al.; A generally nonlethal Salmonella typhimurium infection in weanling rats produced bacterial myocarditis and myocardial hyperplasia . Myocardial lesions were characterized by focal infiltrates of inflammatory cells (predominantly mononuclear), segmental myocyte necrosis, and incipient fibrosis . Although bacterial infections are infrequently associated with myocarditis, the S . typhimurium infection in young rats produced a new experimental model of diffuse myocardial inflammatory foci . Biochemical changes in the myocardium included great increases in total myocardial contents of protein (23%), RNA (39%) and DNA (43%) and several lipid fractions (35-55%) as well as in tissue activities of acid hydrolases, such as cathepsin D (124%) and beta-glucuronidase (135%), all of which contrasted with the relatively limited areas of histologic involvement (1.5%) . To study the effects of additional stress in this model infection, some rats were exercised by forced running in wheels for 2 hours and others were fasted for 24 hours before samples were obtained . The short period of forced exercise in this infection caused an additional increase of myocardial protein content (47%) but with no additional change in histology . The expected fasting-induced degradation of protein as well as an infection-associated increase in myocardial lipids were each prevented when rats were fasted during ongoing acute infection . Protein degradation, as reflected by heightened acid hydrolase activities, seemed to occur at a similar rate regardless of other stresses, whereas the rate of myocardial protein synthesis appeared to be alterable. EMBO J, 1984 Jul, 3(7), 1587 - 93 Molecular cloning, sequencing, and expression of the crr gene: the structural gene for IIIGlc of the bacterial PEP:glucose phosphotransferase system; Nelson SO et al.; The phosphoenolpyruvate:glucose phosphotransferase system (PTS) of Salmonella typhimurium is involved both in glucose transport and in the regulation and synthesis of adenylate cyclase and several transport systems . The crr gene has been implicated in this regulating mechanism . A 9.6-kb segment of the S . typhimurium chromosome containing the crr gene was cloned in pAT153 . The cloned fragment also complemented cysA mutations but did not contain a functional pts operon which is closely linked to the crr gene and codes for two enzymes of the PTS . Although cysA and crr have been reported to be located on opposite sides of ptsHI, our results suggest that the correct gene order is cysK-ptsHI-crr-cysA . Expression of crr plasmids in a maxicell system yielded two proteins which reacted with specific anti-serum against IIIGlc . The apparent mol . wts . in SDS-polyacrylamide gels were 20 000 and 21 000, the former corresponding to the major band of purified IIIGlc . Both forms were also observed in bacterial extracts and purified IIIGlc . The crr gene was localized on a 1-kb EcoRI-EcoRV fragment of the 9.6-kb insert and sequenced . It codes for a single protein (18 556 D) containing 169 amino acid residues and identified as IIIGlc. J Biol Chem, 1984 Jun 25, 259(12), 7719 - 25 Sites of methyl esterification and deamination on the aspartate receptor involved in chemotaxis; Terwilliger TC et al.; The receptors involved in bacterial chemotaxis are post-translationally modified by specific enzymes which catalyze the deamination of glutaminyl residues and the methyl esterification and demethylation of glutamyl residues . In this work we identify the sites of these covalent modifications on the aspartate receptor from Salmonella typhimurium . These were identified using the properties of the Staphylococcus aureus V8 protease which cleaves peptide bonds following glutamyl but not glutaminyl residues . We show here that bonds following methyl-esterified glutamyl residues are also resistant to the protease . A comparison of the fragments obtained after V8 protease cleavage of methyl-esterified (or deaminated) peptides with the fragments from the corresponding unmodified peptides immediately yields the sites of modification . Three of the four methyl-esterified glutamyl residues are located near the middle of the receptor amino acid sequence; one of these is synthesized as a glutaminyl residue and is deaminated by the esterase to form a glutamyl residue . The fourth site of methyl esterification is located near the carboxyl terminus . All four sites occupy analogous positions in a well-conserved arrangement of residues which may form a binding site for the esterase and the methyltransferase. Eur J Biochem, 1984 Jun 15, 141(3), 579 - 83 The hemolytic effect of Salmonella typhi Ty 2 porins; Calderon I et al.; Two outer membrane proteins of Salmonella typhi Ty 2 were extensively co-purified . According to their migration in dodecylsulfate/polyacrylamide gel electrophoresis and solubility characteristics, these proteins are homologous to the 35-kDa and 36-kDa porins found in Salmonella typhimurium . A porin homologous to the 34-kDa one has not been found in S . typhi Ty 2 . A critical step in the purification of porins is heating at 100 degrees C in 2% sodium dodecyl sulfate before Sephadex gel filtration . The absence of detergent in aqueous suspensions enhances porin aggregation, these aggregations inducing human red cell lysis . Porins obtained by an alternative procedure consisting of heating at 60 degrees C instead of 100 degrees C were also hemolytic . Using nanomolar concentration of porins a strong influence of temperature on the hemolytic effect was observed . Porin-induced hemolysis was inhibited with anti-porin serum, as well as by a treatment with phenylglyoxal, which reacts with the arginine residues of proteins . The membrane-disrupting ability of porins aggregates might explain some pathogenic characteristics of gram-negative bacterial infections. Microbiol Sci, 1984 Jun, 1(3), 69 - 72 Phylogeny of strains of Salmonella typhimurium; Old DC; The combined use of biotyping and phage typing has been used to identify major clones of Salmonella typhimurium . Recombination studies among isolates of different clones indicate their genetic relatedness and allow construction of a phylogenetic tree showing relationships among major biotypes . Possible lines of their descent from a common archetypal ancestor are discussed. Appl Environ Microbiol, 1984 Jun, 47(6), 1355 - 7 Mutagenicity of tetramic mycotoxin cyclopiazonic acid; Sorenson WG et al.; Cyclopiazonic acid was shown to be mutagenic to Salmonella typhimurium<