Vet Microbiol, 1994 Jun, 40(3-4), 271 - 82 Genomic lineage of Salmonella enterica serovar Dublin; Olsen JE et al.; Thirty five strains of the host adapted Salmonella serotype Dublin (S . Dublin) have been characterized by IS200 patterns, ribotyping, pulsed-field gel electrophoresis (PFGE), restriction fragment polymorphism after hybridization with five randomly cloned DNA-fragments of S . enteritidis (RFLP), and plasmid profiling in order to divide the strains into 'genomic lines' . For comparison, 20 other strains of 9 different group-D serotypes were included . The IS200 patterns were identical in all strains of S . Dublin examined . These patterns were different from those observed in other group-D Salmonella with the exception of one strain S . Enteritidis phage type 11 and a strain of S . Rostock . The insertion element IS200 was not detected in strains of S . Dar-es-Salam, S . (II) 9,12:z:-, and S . Panama . RFLP, based on probing with five random cloned chromosomal fragments gave the same pattern in all strains except for one isolate from the UK . This strain was also found to have an unique PFGE pattern and ribotype . Among the remaining strains, three different PFGE patterns and 7 different ribotypes were observed . Based on all four typing methods, 8 different 'genomic lines' of S . Dublin were identified . The same grouping could be obtained from the use of ribotyping alone, but PFGE and RFLP were found to provide valuable information on possible relationships between ribotypes . Seven different plasmid profiles and a group of strains without plasmids were observed . In several cases, the same plasmid profile was shown to be present in more than one 'genomic line'. Mol Microbiol, 1994 Jun, 12(6), 865 - 71 Rhs elements of Escherichia coli: a family of genetic composites each encoding a large mosaic protein; Hill CW et al.; The Rhs family comprises a set of composite elements found in the chromosomes of many natural Escherichia coli strains . Five Rhs elements occur in strain K-12 . The most prominent Rhs component is a giant core open reading frame (core ORF) whose features are suggestive of a cell surface ligand-binding protein . This hypothetical protein contains a peptide motif, xxGxxxRYxYDxxGRL(I or T)xxxx, that is repeated 28 times . A similar repeated motif is found in a Bacillus subtilis wall-associated protein . The Rhs core ORFs consist of two distinct parts: a large N-terminal core that is conserved in all Rhs elements, and a smaller C-terminus that is highly variable . Distinctive G+C contents of Rhs components indicate that the elements have a recent origin outside the E . coli species, and that they are composites assembled from segments with very different evolutionary histories . The Rhs cores fall into three sub-families that are mutually more than 20% divergent . Downstream of the core ORF is a second, much shorter ORF . Like the adjacent core extension, these are highly variable . In most examples, the hypothetical product of this ORF has a candidate signal sequence for transport across the cytoplasmic membrane . Another Rhs component, the 1.3 kb H-rpt, has features typical of insertion sequences . Structures homologous to H-rpt have been detected in other bacterial genera, such as Vibrio and Salmonella, where they are associated with loci that determine O-antigen variation. Int J Pancreatol, 1994 Jun, 15(3), 229 - 30 Necrotizing acute pancreatitis induced by Salmonella infection; Andren-Sandberg A et al.; A case of salmonellosis complicated by hemorrhagic pancreatitis is presented . It is emphasized that removal of the gallbladder when stones are present is mandatory in sepsis induced by salmonellosis in the bile-pancreatic region, in spite of modem antibiotic treatment. J Antimicrob Chemother, 1994 Jun, 33(6), 1173 - 89 Quinolone resistance in veterinary isolates of Salmonella; Griggs DJ et al.; Twenty-seven nalidixic acid-resistant (MIC > or = 256 mg/L) isolates of salmonella from veterinary sources were also less susceptible to fluoroquinolones (range of MICs of ciprofloxacin, 0.12-2 mg/L) . Six isolates were cross-resistant to one or more chemically unrelated antibacterial agents . The concentrations of enrofloxacin that inhibited DNA synthesis by 50% were similar to the MIC values for 23 of 27 isolates, suggesting a mutation in gyrA . Insertion of pNJR3-2 (gyrA) in nine of 20 isolates increased susceptibility to quinolones, suggesting that resistance was due to mutation in gyrA . Five of 27 isolates had reduced levels of accumulation of enrofloxacin . Two of the five also had increased susceptibility to quinolones when pNJR3-2 was introduced . None of the outer membrane protein profiles of the resistant isolates differed from those of sensitive control strains . Three of 27 isolates had differences in lipopolysaccharide profiles compared to control strains . Although the MIC of ciprofloxacin was less than the recommended UK break point concentrations for most isolates, the increasing incidence of quinolone-resistance in salmonella from veterinary sources is a matter of concern. Clin Invest Med, 1994 Jun, 17(3), 212 - 7 Serologic testing for reactive arthritis; Thomson GT et al.; The objective of the study was to determine the sensitivity and specificity of quantitative serum antibody response to Salmonella enteritidis lipopolysaccharide (LPS) as a diagnostic test for post-Salmonella reactive arthritis (ReA) . In a single food-source outbreak of Salmonella enteritidis, serum was collected from dysenteric individuals with and without ReA at 6, 12, and 24 months post infection . Serum was also collected from control patients with no prior exposure to Salmonella infection . Quantitative measurements of isotypic antibodies to Salmonella enteritidis LPS were performed by an ELISA . Sensitivity and specificity of quantitative isotypic antibody levels over time were plotted on receiver operator characteristic (ROC) curves . Serum IgG and IgA anti-LPS were found to be present in higher levels in the ReA patients than in controls . Using the optimal cutoff of 0.10 selected from an ROC curve, IgG anti-LPS is 88% sensitive and 94% specific, and IgA anti-LPS is 75% sensitive and 100% specific . We conclude that IgA anti-LPS is both sensitive and specific in distinguishing prior exposure to Salmonella LPS in ReA patients compared to unexposed controls. FEMS Immunol Med Microbiol, 1994 Jun, 9(1), 49 - 54 Efficacy of tumor necrosis factor alpha and eicosanoid inhibitors in experimental models of neonatal sepsis; Mancuso G et al.; The potential role of tumor necrosis factor alpha (TNF alpha) and eicosanoids in the pathogenesis of experimental neonatal sepsis models was investigated . Lethality was induced in neonatal rats by administration of heat killed group B streptococci (GBS, 7 mg kg-1 intracardially) or Salmonella enteritidis endotoxin (0.35 mg kg-1 intracardially) . The relative efficacy of six compounds with putative TNF alpha and eicosanoid inhibitory actions were tested . These were: ibuprofen (3 and 20 mg kg-1), a cyclo-oxygenase inhibitor; CGS85515 (30 mg kg-1), a lipoxygenase inhibitor; LY203647 (30 mg kg-1), a leukotriene D4 receptor antagonist; pentoxifylline (10, 50 and 100 mg kg-1), a TNF inhibitor; cloricromene (2 and 10 mg kg-1), a thromboxane A2 synthetase inhibitor with TNF alpha inhibitory actions; and SKF86002 (2.5, 5, 10 and 20 mg kg-1), a dual cyclo-oxygenase/lipoxygenase inhibitor with TNF alpha inhibitory activity . Pentoxifylline, cloricromene and SKF86002, when given intraperitoneally 2 h before challenge, produced 45, 52 and 61% reductions, respectively, in plasma levels of TNF alpha at 2.5 h post-injection with killed GBS (P < 0.05) . On the contrary, pretreatment with ibuprofen, CGS85515 or LY203647 did not significantly affect TNF alpha levels . All compounds significantly attenuated the lethality by killed GBS and S . enteritidis endotoxin . These data suggest that TNF alpha and eicosanoids contribute to the pathogenesis of shock induced by killed GBS and endotoxemia. Asian Pac J Allergy Immunol, 1994 Jun, 12(1), 27 - 37 Molecular cloning and expression of Salmonella paratyphi A 52 kDa specific protein gene; Korbsrisate S et al.; Monoclonal antibodies (MAbs) specific to Salmonella paratyphi A have been established by our group in 1989 . These MAbs were proven to be species-specific for 52 kDa protein of S . paratyphi A but the nature of this protein is unknown . However, our group have proved that the 52 kDa protein which is specific to S . typhi was flagellin . This present study has characterized the 52 kDa protein of S . paratyphi A and identified its encoded gene . The plasmid containing the specific 52 kDa antigen gene was cloned from the S . paratyphi A genome, herein designated pSKA-4 . Partial nucleotide sequences from this clone was analysed by computer program and found to be phase 1-a flagellin gene of S . paratyphi A . In addition, the nucleotide sequence analysis from such clone also showed that the structural gene for phase 1 flagellin has amino acid sequences conserved at the terminal whereas the central region is variable among Salmonella spp . Therefore, the central portion of flagellin which highly polymorphic in amino acid sequences would be the most specific to S . paratyphi A, thus, should be used as specific antigen for developing specific diagnosis of S . paratyphi A infection . Using the PCR technique, an expression plasmid containing the antigen gene producing only the variable region in the central portion of flagellin from S . paratyphi A, namely pSKA-7, has been established . The recombinant protein produced by the established plasmid has a MW 33.5 kDa as detected by immunoblotting using specific MAbs . Further study by using this specific flagellin protein for immunodiagnosis of S . paratyphi A infection is being carried out in our laboratory. Asian Pac J Allergy Immunol, 1994 Jun, 12(1), 21 - 5 Fusion protein of Salmonella typhi flagellin as antigen for diagnosis of typhoid fever; Sukosol T et al.; We previously established the specific 52 kDa antigen of Salmonella typhi, detected by our monoclonal antibodies, which was a flagellin protein . Comparison of the nucleotide sequences of phase-1 flagellin of Salmonella species available through GenBank database showed high homology at both ends of the genes with lower degree of homology in the middle portion which contained the antigenically variable regions . Thus, proteins from the central regions of flagellin genes should be species specific and could be used as specific antigens for the immunodiagnostic tests . In this report, recombinant protein derived from the central region of S . typhi flagellin was produced as a fusion protein with glutathione-S-transferase . This fusion protein was used as specific S . typhi antigen for the immunodiagnostic test to detect IgM antibodies in sera using enzyme-linked immunosorbent assay . The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of this test were 53.5, 98.0, 91.5, 82.1 and 92.4%, respectively. Eur J Epidemiol, 1994 Jun, 10(3), 345 - 7 In vitro activity of eight antibiotics against Salmonella and Shigella species; Arman D et al.; The minimum inhibitory concentrations (MICs) of ampicillin, chloramphenicol, trimetoprimsulfamethoxazole (TMP-SMX), ofloxacin, ciprofloxacin, pefloxacin, enoxacin and fleroxacin for Salmonella spp . (n = 72) and Shigella spp . (n = 52) were established . S . typhi isolates were all susceptible to all of the antibiotics tested . In non-typhi salmonellae and Shigella spp., resistance to ampicillin, chloramphenicol and TMP-SMX with various rates were encountered, but all isolates were susceptible to fluoroquinolones. Southeast Asian J Trop Med Public Health, 1994 Jun, 25(2), 324 - 7 Detection of salmonellae in hen eggs in Thailand; Saitanu K et al.; Two thousand four hundred and ninety eggs were collected from retail markets in 6 provinces and from laying hen farms in 3 provinces . Eggs were pooled in groups of 3 to obtain 830 samples for testing . Isolation of salmonellae was made from both egg shell and egg contents . Eggs from retail markets were contaminated with salmonellae on egg shells (13.2%) and in egg contents (3.9%) . Three (0.4%) samples yield positive both on egg shells and in egg contents . Of the 86 samples from laying hen farms, salmonellae were found on egg shells and in egg contents, 3.5% and 1.2%, respectively . From the 134 strains tested, twenty-four serotypes were confirmed . Salmonella cerro, S . amsterdam and S . typhimurium were predominantly encountered, 4.8%, 4.3% and 1.4%, respectively . Only two samples were contaminated with S . enteritidis, one each from open market and laying hen farm, one on egg shells and the other in egg content respectively. Circ Shock, 1994 Jun, 43(2), 64 - 70 Beneficial effects of extracellular glutathione against endotoxin-induced liver injury during ischemia and reperfusion; Liu P et al.; The potential beneficial effect of hepatocellular glutathione against inflammatory liver damage was investigated in a model of endotoxin-enhanced ischemia-reperfusion injury . Animals were subjected to 20 min of hepatic ischemia, followed by 4 hr of reperfusion . The injection of 0.5 mg/kg Salmonella enteritidis endotoxin potentiated liver injury and the postischemic oxidant stress, as indicated by increased plasma levels of glutathione disulfide . Depletion of hepatic glutathione levels by > 90% with phorone and inhibition of glutathione synthesis with buthionine sulfoximine further increased liver injury in this model, as indicated by enhancement of plasma alanine aminotransferase activities from 2,234 +/- 122 U/L to 4,024 +/- 282 U/L . Continuous infusion of a glutathione (GSH) solution in GSH-depleted animals (22 mumol/kg/hr) attenuated reperfusion injury by 55% . In vitro experiments demonstrated the capability of GSH to react rapidly with reactive oxygen species, such as hydrogen peroxide (H2O2) and hypochlorous acid (HOCl) . Only H2O2 oxidized GSH quantitatively to its disulfide; HOCl oxidized GSH to higher oxidation states . These data support the hypothesis that the enhanced release of hepatocellular GSH functions as a defense mechanism against reactive oxygen species generated by inflammatory cells during endotoxemia and reperfusion . This internal defense system of the liver may be of general importance in preventing, or at least limiting, liver damage by reactive oxygen generated in particular by Kupffer cells during their physiological function to remove gut-derived endotoxin and bacteria. Zentralbl Bakteriol, 1994 Jun, 281(1), 30 - 7 Influence of the glucose concentration on the yield of biomass and lipopolysaccharide in Salmonella cultures; Schlecht S et al.; Two Salmonella S-forms and two R-mutants were cultivated in complex medium supplemented with different amounts (0.5-3%) glucose . Cultivation was performed batchwise in a fermentor under aerobic conditions . With all strains investigated, the yield of bacterial mass increased with increasing concentration of glucose . In the case of three strains, the % LPS content of bacteria also increased, thus achieving the aim of this investigation . The synthesis of bacterial mass and LPS did not proceed in parallel and differed from strain to strain . At optimal glucose concentration, the yield of LPS could be increased up to 250% . The chemical composition of the LPS was independent of the glucose concentration . The individual strains exhibited an identical composition with regard to lipid A and polysaccharide when cultured at different glucose concentrations . The uniformity of the molecular distribution of LPS could also be confirmed by SDS-polyacrylamide gel electrophoresis . In the S-form LPSs, also the proportion of the unsubstituted R-form LPS was not affected by the glucose concentration in the culture medium . The present results demonstrate that optimisation of the cultivation conditions with respect to the glucose concentration of the medium would be of advantage especially for Salmonella strains that are cultivated frequently. Mutat Res, 1994 Jun, 321(4), 241 - 51 Mutagenicity of ethanolic extracts of used acrylic dentures; Parisis DM et al.; The in vivo physicochemical sorption of mutagenic substances onto acrylic polymers was investigated in worn acrylic dentures . Thus, ethanolic extracts of acrylic dentures from 41 of a total of 69 human donors (60%), were found mutagenic in the standard plate incorporation Salmonella mutagenicity test against either TA98 or TA100 strains . Denture extracts from smokers produced mutagenicity more often than the ones from non-smokers (75% vs . 45%, P 0.01) . Mutagenicity was preferentially directed against TA98 (TA98:TA100 = 2.9:1, P < 0.0005) . Predilection for TA98 was more pronounced in denture extracts from non-smokers (4.7:1) than from smokers (2.0:1) . When direct mutagenicity was observed, it was reduced by the rat-liver S9 . Induced mutant yields were 6.1 +/- 3.9 and 7.0 +/- 8.9 times higher than the spontaneous for TA98 and TA100 respectively (smokers, 50-cm2 denture surface area eq./plate+S9) . Denture extracts from smokers induced higher levels of mutation than the ones from non-smokers (TA98 + S9, smoker:non-smoker = 2:1, P < 0.01) . Mutagenicity was associated with longer periods of denture usage (P 0.007) . Thus, denture poly(methyl methacrylate) base material can adsorb mutagenic substances, possibly from diet and tobacco, which are extractable by ethanol . Theoretically, the in situ alcoholic desorption and recirculation of carcinogenic mutagens may have a contributory role in certain cases of intra-oral and upper alimentary tract carcinogenesis. Mutat Res, 1994 Jun, 321(4), 229 - 39 Mutagenicity of tryptophan photoproducts in the Ames Salmonella assay; Sjogren M et al.; During the photolysis of tryptophan a large number of products is formed . In this study, aqueous solutions of tryptophan were irradiated by ultraviolet light during 5, 20 or 40 h . Each of the irradiated batches was divided into two aliquots, which were freeze-dried or extracted with chloroform . For each batch the latter extract was subsequently divided into a purified chloroform extract and a methanol extract . Aliquots of the purified chloroform extracts were fractionated and pooled, peakwise, into seven fractions . A recombined sample was also constructed . All extracts and samples were tested for mutagenicity using the standard Ames Salmonella assay . The results indicate an exposure time dependent increase in mutagenicity of the extracts, as seen with tester strain TA100 both with and without metabolic activation . The mutagenicity of the freeze-dried extracts well approximated the mutagenicity of the chloroform extracts, indicating that most mutagenicity can be extracted with chloroform . With the fractions the highest mutagenic responses were seen in the late, i.e., less lipophilic fractions . This response pattern seen in TA98 and TA100, mainly with S9 activation, was in contrast to the response of TA102 without S9, which was highest to the more lipophilic fractions . On a fraction level, no general exposure dependent increase of mutagenicity was observed . The results also show that photooxidation of tryptophan gives rise to a different spectrum of products compared to pyrolysis . Both processes result in compounds with strong biological effects . Photooxidation results in compounds with low genotoxicity and high Ah receptor affinity while pyrolysis generates compounds with high genotoxicity and low or no Ah receptor affinity. Mutat Res, 1994 Jun, 321(4), 213 - 8 Genotoxicity of 1,3-dithiane and 1,4-dithiane in the CHO/SCE assay and the Salmonella/microsomal test; Lee H et al.; 1,3-Dithiane and 1,4-dithiane are the sulfur-containing Maillard reaction products (MRPs) which have been found in boiled beef extracts . In this study the genotoxicity of these products was examined using the Salmonella/microsomal test and the CHO/SCE assay . 1,3-Dithiane showed a potent direct-acting mutagenicity toward S . typhimurium TA98 and TA100, but 1,4-dithiane had a lower mutagenicity toward both tester strains . Both compounds were shown to be non-mutagenic with hepatic metabolic activation with the exception of 1,3-dithiane toward strain TA100 . To compare the mutagenic potential of 1,3-dithiane and 1,4-dithiane with other types of MRPs, 24 MRPs were examined for their mutagenicity to S . typhimurium TA98 and TA100 in the presence or absence of S9 mix . 2,6-Dimethylpyrazine, furan, 2-acetylpyrrole, and thiazole were shown to be mutagenic . However, these four MRPs exhibited a lower mutagenicity in TA98 than 1,3-dithiane and 1,4-dithiane . Furthermore, SCE frequencies in CHO cells were very significantly induced by 1,3-dithiane in the absence of S9 mix, but the SCE-inducing capability of 1,3-dithiane was reduced or even disappeared with metabolic activation . 1,4-Dithiane did not significantly induce SCE frequencies in the presence or absence of S9 mix . Thus, we concluded that 1,3-dithiane was a potent mutagenic MRP in the Salmonella/microsomal test, whereas it was a weak SCE inducer in the CHO/SCE assay. Eur J Biochem, 1994 May 15, 222(1), 183 - 94 Preparation and structural analysis of oligosaccharide monophosphates obtained from the lipopolysaccharide of recombinant strains of Salmonella minnesota and Escherichia coli expressing the genus-specific epitope of Chlamydia lipopolysaccharide; Holst O et al.; The lipopolysaccharide of the recombinant strain Salmonella minnesota r595-207 expressing the genus-specific epitope of Chlamydia lipopolysaccharide {Holst, O., Brade, L., Kosma, P . and Brade, H . (1991) J . Bacteriol, 173, 1862-1866} was sequentially de-O- and de-N-acylated by mild hydrazinolysis and treatment with 4 M KOH, respectively . The resulting mixture of compounds was separated by high-performance anion-exchange chromatography and gel-permeation chromatography, yielding four oligosaccharide phosphates two of which were readily identified by their 1H-NMR- and 13C-NMR spectra as alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha-D-Glcp N 1,4'-bisphosphate (tetrasaccharide bisphosphate; Kdo = 3-deoxy-D-manno-octulopyranosonic acid) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha-D- GlcpN 1,4'-bisphosphate (pentasaccharide bisphosphate) {Holst, O., Broer, W., Thomas-Oates, J.E., Mamat, U . and Brade, H . (1993) Eur . J . Biochem . 214, 703-710} . The structures of the other two compounds were established by chemical analysis, NMR spectroscopy, and fast-atom-bombardment mass spectrometry as alpha-Kdo- (2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha-D-GlcpN 1-phosphate (tetrasaccharide 1-phosphate) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha-D- GlcpN 1-phosphate (pentasaccharide 1-phosphate) . alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha/beta- D-GlcpN 4'-phosphate (tetrasaccharide 4'-phosphate) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha/beta-D-GlcpN 4'-phosphate (pentasaccharide 4'-phosphate) were prepared from the 1,4'-bisphosphates isolated from the recombinant strain Escherichia coli F515-207 by treatment with alkaline phosphatase and purification by high-performance anion-exchange chromatography and gel-permeation chromatography . Their structures were characterised by chemical analysis, NMR spectroscopy, and fast-bombardment mass spectrometry. Biochemistry, 1994 May 3, 33(17), 5183 - 92 Solution structure of a trisaccharide-antibody complex: comparison of NMR measurements with a crystal structure; Bundle DR et al.; NMR and crystallography have been used to study antigen conformational changes that occur in a trisaccharide-Fab complex in solution and in the solid state . NOE buildup rates from transferred NOE experiments show that the antigenic determinant of a Salmonella lipopolysaccharide, represented by the trisaccharide methyl glycoside alpha-D-Galp(1-->2 {alpha-D-Abep(1-->3)}- alpha-D-Manp1-->OMe (1), undergoes a protein-induced conformational shift about the Gal-->Man glycosidic linkage when it is bound by a monoclonal antibody in aqueous solution . The same trisaccharide was crystallized with Fab, and a solved structure at 2.1-A resolution revealed that the conformation of the trisaccharide ligand was similar to that seen in a dodesaccharide-Fab complex {Cygler et al . (1991) Science 253, 442-445), where the Gal-Man linkage also experienced a similar conformational shift . Distance constraints derived from the TRNOE buildup curves are consistent with two bound trisaccharide conformations, one of which correlates with the ligand conformation of the crystalline Fab-trisaccharide complex . In this bound conformation, short interatomic distances between Abe O-2 and Gal O-2 permit an oligosaccharide intramolecular hydrogen bond . Despite its relatively low energy, a preponderance of this conformer could not be detected in aqueous or DMSO solutions of free trisaccharide by either 1H or 13C NMR experiments . In DMSO, a different intramolecular hydrogen bond between Abe O-2 and Man O-4 was observed due to a solvent-induced shift in the conformational equilibria (relative to aqueous solution) . Molecular modeling of the trisaccharide in the binding site and as the free ligand suggested that the protein imposes an induced fit on the antigen, primarily resulting in a shift of the Gal-Man phi torsional angle . This reduces the interproton separation between Abe H-3 and Gal H-1 with a marked increase in the intensity of the previously weak NOEs between the protons of the noncovalently linked galactose and abequose residues . The impact of the conformational shift on gross trisaccharide topology is sufficiently small that binding modes inferred from functional group replacements are not impaired. Biochemistry, 1994 May 3, 33(17), 5172 - 82 Molecular recognition of a Salmonella trisaccharide epitope by monoclonal antibody Se155-4; Bundle DR et al.; The binding site of monoclonal antibody Se155-4, which has been the object of successful crystallographic and antibody-engineering studies, is shown by solid-phase immunoassays to be complementary to a branched trisaccharide, alpha-D-Galp(1-->2) {alpha-D-Abep(1-->3)}-alpha-D-Manp(1, rather than to the tetrasaccharide repeating unit alpha-D-Galp(1-->2) {alpha-D-Abep(1-->3)}-alpha-D-Manp(1-->4) alpha-L-Rhap(1- of the bacterial antigen . Specificity for the 3,6-dideoxy-D-xylo-hexose (3,6-dideoxy-D-galactose) epitope present in Salmonella paratyphi B O-antigens was ensured by screening hybridoma experiments with glycoconjugates derived from synthetic oligosaccharides . Detailed epitope mapping of the molecular recognition by modified and monodeoxy oligosaccharide derivatives showed that complementary surfaces and three antibody-saccharide hydrogen bonds are essential for full binding activity . Both hydroxyl groups of the 3,6-dideoxy-D-galactose residue were obligatory for binding and consistent with the directional nature of their involvement in carbohydrate-protein hydrogen bonds; related tetrasaccharides built from the isomeric 3,6-dideoxyhexoses, 3,6-dideoxy-D-glucose, paratose, and 3,6-dideoxy-D-mannose, tyvelose were not bound by the antibody . Titration microcalorimetry measurements were consistent with the hydrogen-bonding map inferred from the crystal structure and suggest that the displacement of water molecules from the binding site accounts for the favorable entropy that accompanies binding of the native trisaccharide determinant . The protein sequences determined for the antibody VL and VH domains reveal somatic mutation of the VL germ line gene, implying that this antibody-binding site results from a mature antibody response. Am J Trop Med Hyg, 1994 May, 50(5), 608 - 11 Detection of the H1-j strain of Salmonella typhi among Korean isolates by the polymerase chain reaction; Song JH et al.; Salmonella typhi, the etiologic agent of typhoid fever, typically has only a phase-1 flagellar antigen, H1-d (fliC) . While most strains of S . typhi have H1-d antigen, 10-20% of Indonesian isolates have been reported to possess H1-j antigen instead . To investigate the presence H1-j strains of S . typhi isolates in Korea, where typhoid fever is still a common infectious problem, we used the polymerase chain reaction (PCR) with a pair of oligonucleotides primers that specifically amplified the flagellin gene of S . typhi . Of 375 isolates of S . typhi tested, only one was shown to possess the H1-j antigen, which was shown by the presence of a 1,269-basepair fragment on agarose gel electrophoresis after the PCR . The isolate with the H1-j antigen was cultured from a Korean-Indonesian man who was already symptomatic in Indonesia and was thought to be an Indonesian strain . Because 375 strains tested in this study were collected from cases with typhoid fever in different regions of Korea during the period from 1986 to 1991, it could be concluded that the mutation rate to j antigen is negligible among S . typhi endemic in Korea. Carcinogenesis, 1994 May, 15(5), 837 - 43 High promutagen activating capacity of yeast microsomes containing human cytochrome P-450 1A and human NADPH-cytochrome P-450 reductase; Sengstag C et al.; Yeast Saccharomyces cerevisiae strains have been constructed that co-express cDNAs coding for the human cytochrome P-450 enzymes CYP1A1 or CYP1A2 in combination with human NADPH-cytochrome P-450 reductase (oxidoreductase) . Microsomal fractions prepared from the strains were able to efficiently activate various drugs to Salmonella mutagens . These experiments demonstrated that a functional interaction occurred between the respective human enzymes in the yeast microsomes . For every drug tested, the microsomes containing CYP enzymes and oxidoreductase were 2- to 4-fold better in activation than the corresponding microsomes that contained CYP alone . Interestingly, co-expression of CYP1A2 with oxidoreductase resulted in a decrease of 7-ethoxyresorufin-O-deethylase activity, a problem which is related to this specific substrate . Using the microsomes, it was demonstrated that aflatoxin B1 was activated to a mutagen not only by CYP1A2 but also by CYP1A1 . In contrast, benzo{a}pyrene was exclusively activated by CYP1A1 whereas CYP1A2 was inactive . The drug 3-amino-1-methyl-5H-pyrido{4,3-b}indole (Trp-P-2) was activated by CYP1A2 and to a lesser extent by CYP1A1 . A strong substrate specificity was observed with the two structurally related heterocyclic arylamines 2-amino-3,4-dimethylimidazo{4,5-f}quinoline (MeIQ) and 2-amino-3,4-dimethylimidazo{4,5-f}quinoline (MeIQ) and 2-amino-3,8-dimethylimidazo{4,5-f}quinoxaline (MeIQx) . MeIQx was activated efficiently by both CYP enzymes, whereas MeIQ was only activated by CYP1A2 and not by CYP1A1 . The fact that microsomes from vector transformed control strains were unable to activate any of the drugs studied underlines the suitability of these microsomes for metabolic studies . Moreover, the presence of suitable marker genes in the yeast strains will enable us to study mitotic recombination and gene conversion events induced by drugs that require metabolic activation. Am J Public Health, 1994 May, 84(5), 859 - 60 Outbreak and sporadic egg-associated cases of Salmonella enteritidis: New York's experience; Morse DL et al.; Since 1985, egg-associated Salmonella enteritidis has emerged as a major cause of foodborne disease . New York State has been especially affected, with 47 documented egg-associated S enteritidis outbreaks involving 2279 cases and 10 deaths . Individual case reports of salmonella have also increased 56%, and sporadic cases of S enteritidis have been shown to be associated with egg consumption . Further educational and regulatory activities are needed to control this continuing public health problem. Infect Immun, 1994 May, 62(5), 1961 - 7 Altered antigen-presenting capacity of human monocytes after phagocytosis of bacteria; Pryjma J et al.; The antigen-presenting and accessory functions of monocytes were studied after phagocytosis of bacteria . Peripheral blood monocytes isolated from mononuclear cells by counterflow elutriation were incubated with suspensions of opsonized bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, or Salmonella enteritidis) under conditions in which at least 80% of the monocytes engulfed microorganisms . Either the cells were pulsed with antigen (purified derivative of tuberculin or tetanus toxoid) and used as antigen-presenting cells for autologous T lymphocytes or the accessory function of the cells was examined in pokeweed mitogen-activated cultures of T cells . It has been found that phagocytosis of bacteria by monocytes reduces their ability to trigger antigen- and mitogen-induced proliferation . The reduced proliferative response of T lymphocytes was not due to a change of the kinetics of the response or triggering of suppressor mechanisms . Furthermore, antigen processing was not affected much after phagocytosis of bacteria since antigen-pulsed and paraformaldehyde-fixed cells containing bacteria were comparable to control cells in their antigen-presenting capacity . This phenomenon was observed after phagocytosis of both living and dead bacteria and was not correlated to the viability of monocytes, which were more affected after phagocytosis of living bacteria than of dead ones . As a result of phagocytosis of bacteria, reduced expression of CD54, CD14, and HLA-DQ, variable reduction of HLA-DP, CD58, and CD64, and reduced viability of monocytes were observed . In conclusion, phagocytosis of bacteria by monocytes affects their antigen-presenting and accessory functions presumably because of changes in the expression of molecules essential for monocyte-T-cell interactions and reduction of their viability. Infect Immun, 1994 May, 62(5), 1705 - 9 Free hydroxyl groups are not required for endotoxic activity of lipid A; Tanamoto K; Previous studies demonstrated that lipid A from Salmonella abortusequi loses its B-cell mitogenicity for murine spleen cells as a result of the introduction of succinyl residues on hydroxyl groups and that the inactivated lipid A specifically antagonizes the mitogenicity of endotoxin . Hypothesizing that the hydroxyl groups are essential both for its biological activity and for producing nontoxic preparations having antagonistic activity, I tested the role of the hydroxyl groups in its activities by using well-characterized biologically active lipid A preparations synthesized chemically (Escherichia coli and Salmonella types 506 and 516, respectively) by the introduction of either succinyl or acetyl residues at the hydroxyl groups of each of these lipid A preparations . However, the biological activities of neither lipid A preparation were reduced at all after succinylation; in fact, succinylated 516 became much more potent than the original molecule with respect to most activities tested, i.e., lethal toxicity, Limulus gelation activity, and the induction of tumor necrosis factor release . On the other hand, when the hydroxyl groups were replaced with acetyl residues, the lethality and tumor necrosis factor-inducing activity of both lipid A preparations were decreased, whereas their Limulus gelation activity was increased . Mitogenicity was not affected much by the chemical modifications of either lipid A preparation . These findings indicate that although the residues introduced into the free hydroxyl groups of lipid A modulate its activities, the hydroxyl groups in lipid A need not exist in free form. Infect Control Hosp Epidemiol, 1994 May, 15(5), 311 - 4 Foodhandler-associated Salmonella outbreak in a university hospital despite routine surveillance cultures of kitchen employees; Khuri-Bulos NA et al.; OBJECTIVE: To describe an outbreak of salmonella food poisoning that probably was due to contamination of mashed potatoes by a foodhandler, which occurred despite a policy for routine surveillance stool cultures of kitchen employees . DESIGN: A case control study of 223 individuals who ate the lunch meal on September 23, 1989, at the Jordan University Hospital (JUH) cafeteria . SETTING: Tertiary care university hospital in Amman, the capital of Jordan . PATIENTS: Individuals who developed loose stool or vomiting 6 to 72 hours after eating the lunch meal of September 23, 1989, at the JUH cafeteria . RESULTS: Of 619 individuals, 183 fit the case definition (attack rate, 19.6%); 150 were employees, 26 were inpatients, and seven were visitors . Twelve other employees became sick 4 to 6 days later and probably were infected secondarily . The incubation period ranged from 16 to 72 hours in 183 instances . Symptoms included diarrhea (88%), fever (71%), abdominal pain (74%), dehydration (34%), and bloody stool (5%) . Eighty-four were hospitalized . Cultures of eight food items were negative, but stool culture on 90 of 180 patients and 11 of 61 kitchen employees yielded Salmonella enteritidis group D . A cohort study of 223 individuals revealed a food-specific attack rate of 72% for the steak and potato meal and 18% for the rice and meat meal (RR, 4; CI95, 2.62 to 6.24; P < 0.01) . Stratified analysis of the steak and potato meal revealed that the potatoes were implicated most strongly (RR, 1.93; CI95, 1.42 to 2.64; P < 0.01) . Cultures were obtained from all kitchen employees, and 11 of 61 grew Salmonella enteritidis group D . One asymptomatic, culture-positive employee prepared the mashed potatoes on September 23 . All of these employees had negative stool cultures 3 months earlier . CONCLUSION: This outbreak probably was caused by massive contamination of mashed potatoes by the contaminated hands of the foodhandler . Routine stool culture of foodhandlers is not cost-effective and should not be used as a substitute for health education and proper hygienic practices. Int J Food Microbiol, 1994 May, 22(2-3), 201 - 6 Evaluation of motility enrichment on modified semisolid Rappaport-Vassiliadis medium (MSRV) and automated conductance in combination with Rambach agar for Salmonella detection in environmental samples of a milk powder factory; Joosten HM et al.; The efficacy of motility enrichment on modified semisolid Rappaport-Vassiliadis medium (MSRV) and an automated conductance method for the detection of Salmonella in environmental samples was evaluated . Two hundred and ten environmental samples from unrestricted areas of a milk powder factory, 49 of which were artificially contaminated with Salmonella infantis, were examined . From exactly 100 samples Salmonella could be isolated . With the conventional (ISO-DIS 6579) method a 100% score was obtained, whereas the MSRV method gave 82 positive results . The conductance method permitted the detection of Salmonella in only 66 samples . The use of Rambach agar improved isolation efficiency of Salmonella from enrichment broths. Int J Food Microbiol, 1994 May, 22(2-3), 127 - 40 Potential growth and control of Salmonella in Hispanic type soft cheese; Kasrazadeh M et al.; This study evaluated the growth and control of Salmonella serotypes in a soft Hispanic type cheese (Queso Fresco) . Cheese was made in the laboratory using a commercial procedure and after inoculation it was stored under vacuum at temperatures ranging from 6 to 30 degrees C . The minimum temperature that allowed growth of Salmonella was 8 degrees C . Accumulated data from the growth studies were used for the development of models relating the square root of 1/LT (LT = lag time) and specific growth rate to temperature . The effect of selected antimicrobials (potassium sorbate, sodium benzoate and sodium lactate) on the growth and survival of Salmonella in milk and in cheese at various storage temperatures was also examined . Addition of sodium benzoate (0.3%) to cheese (pH 6.6) or addition of potassium sorbate (0.3%) to cheese (pH 6.0) made from milk, which was acidified to pH 5.9 with propionic acid, had a significant impact on delaying or preventing growth of the pathogen. Boll Chim Farm, 1994 May, 133(5), 328 - 38 Antimicrobial and genotoxic properties of quinoline derivatives; Zani F et al.; We continued our research into the biological properties of quinoline derivatives . Newly synthesized 8-sulfonylquinolines, tested against some representative microbial strains and practically inactive, were also studied for the genotoxic properties . The genotoxicity tests were extended to previously synthesized compounds (some 6-substituted 8-quinolinecarboxylic acids, the amide and some esters of 8-quinolinecarboxylic acid and finally the 8-quinolinecarboxaldehyde and two of its derivatives) . Rec-assay and Salmonella-microsome tests showed several compounds to be genotoxic; the mutagenic activity seems to be modulated by the nature of the substituents . The results obtained are discussed with the aim of explaining possible structure-activity relationships. Am J Vet Res, 1994 May, 55(5), 636 - 42 Adjuvanted subunit vaccines for the control of Salmonella enteritidis infection in turkeys; Charles SD et al.; Liposomes and immunostimulating complexes (ISCOM) are adjuvants that have been known to potentiate the immune response to membrane proteins . Adjuvanted outer membrane proteins (OMP) from Salmonella enteritidis were evaluated for their protective efficacy against S enteritidis infection in turkeys . The adjuvanted vaccines prepared for evaluation were: positive or negatively charged liposomes, lipid-conjugated ISCOM, and mineral oil vaccines . These preparations were compared with that of a whole cell bacterin and protein alone . After vaccination, turkeys were challenge-exposed with a nalidixic acid-resistant strain of S enteritidis . They were monitored for clinical signs of disease, antibody response, bacterial shedding pattern, and clearance of the challenge S enteritidis from internal organs . Results indicated a significantly (P < 0.05) higher antibody response to the positively charged liposomal OMP vaccine, compared with the whole cell bacterin . The antibody response to positively charged liposomal OMP vaccine was greater when a booster dose of this preparation was given . Shedding of S enteritidis was decreased in all vaccinated and challenge-exposed turkeys (P < 0.001) . The tissues from a high percentage (90 to 100%) of birds that received a booster vaccination of the liposomal (+ or -) and ISCOM vaccine were culture-negative for S enteritidis. Mycopathologia, 1994 May, 126(2), 121 - 9 Mutagenic and membranal effect of a phytotoxic molecule isolated from olive leaves parasitized by the fungus Cycloconium oleaginum Cast; Yahiaoui R et al.; A phytotoxic substance (C23H44O3) which is named 'Substance A', was purified from olive leaves infected with Cycloconium oleaginum Cast . The mutagenic effect of this substance was detected using TA 100 and TA 102 strains of Salmonella in the Ames test using Bacillus subtilis strains M45 rec-, H17 rec+ in the rec assay . Another substance manifesting the mutagenic effect was found in the extract from the Cycloconium oleaginum culture . This substance was not detected in the extract from contaminated olive leaves . Substance A increased electrolytes leakage from tissue of olive leaves, thus manifesting its phytotoxicity. Hautarzt, 1994 May, 45(5), 330 - 4 {Snake bite by a poisonous snake . Report of an unusual case}; Gruschwitz MS et al.; We report on a 31-year-old white woman, who was bitten in her right calf by a "spitting cobra" (Neia nigricollis) during a safari in Tansania . Minor initial systemic symptoms such as nausea and vomiting were followed by severe oedematous swelling of the extremity after 2-3 h and demarcation of a 2.75 x 2.75 in . area of necrotic skin . The patient returned to her home country, where 8 days after the snake-bite necrosectomy was performed . Antibiotics, anti-inflammatory agents and local therapy with hydrocolloidal wound dressings were administered . With this therapy the lesion healed completely with minor scarring within 5 months . A new Salmonella strain was isolated from the ground of the ulcer. Poult Sci, 1994 May, 73(5), 648 - 52 Resistance against Salmonella enteritidis cecal colonization in Leghorn chicks by vent lip application of cecal bacteria culture; Corrier DE et al.; Leghorn chicks were treated with cultures of cecal bacteria from adult chickens by crop gavage, upper body spray, or vent lip application on the day of hatch . The chicks were challenged orally with 10(4) Salmonella enteritidis (SE) at 3 d of age and evaluated for SE cecal colonization at 10 d of age . The concentration of volatile fatty acids (VFA) in the cecal contents was determined on the day after culture treatment and at 10 d of age . Compared with controls, SE colonization was significantly decreased in each of the treatment groups . Vent lip application of a single .05-mL drop of cecal bacteria culture resulted in resistance against SE challenge comparable to crop gavage or spray treatment with .5 mL of culture . Resistance to SE challenge was directly associated with the concentrations of total VFA and propionic acid in the cecal contents of the treated chicks on the day after culture treatment . The results indicated that cecal bacteria from adult chickens that increase SE colonization resistance may rapidly become established in the ceca of newly hatched chicks following contact with the vent lips. Immunology, 1994 May, 82(1), 42 - 50 Interaction with extracellular matrix proteins influences Lsh/Ity/Bcg (candidate Nramp) gene regulation of macrophage priming/activation for tumour necrosis factor-alpha and nitrite release; Formica S et al.; The murine resistance gene Lsh/Ity/Bcg regulates activation of macrophages for tumour necrosis factor-alpha (TNF-alpha)-dependent production of nitric oxide mediating antimicrobial activity against Leishmania, Salmonella and Mycobacterium . As Lsh is differentially expressed in macrophages from different tissue sites, experiments were performed to determine whether interaction with extracellular matrix (ECM) proteins would influence the macrophage TNF-alpha response . Plating of bone marrow-derived macrophages onto purified fibrinogen or fibronectin-rich L929 cell-derived matrices, but not onto mannan, was itself sufficient to stimulate TNF-alpha release, with significantly higher levels released from congenic B10.L-Lshr compared to C57BL/10ScSn (Lshs) macrophages . Only macrophages plated onto fibrinogen also released measurable levels of nitrites, again higher in Lshr compared to Lshs macrophages . Addition of interferon-gamma (IFN-gamma), but not bacterial lipopolysaccharide or mycobacterial lipoarabinomannan, as a second signal enhanced the TNF-alpha and nitrite responses of macrophages plated onto fibrinogen, particularly in the Lshr macrophages . Interaction with fibrinogen and fibronectin also primed macrophages for an enhanced TNF-alpha response to leishmanial parasites, but this was only translated into enhanced nitrite responses in the presence of IFN-gamma . In these experiments, Lshr macrophages remained superior in their TNF-alpha responses throughout, but to a degree which reflected the magnitude of the difference observed on ECM alone . Hence, the specificity for the enhanced TNF-alpha responses of Lshr macrophages lay in their interaction with fibrinogen and fibronectin ECM, while a differential nitrite response was only observed with fibrinogen and/or IFN-gamma . The results are discussed in relation to the possible function of the recently cloned candidate gene Nramp, which has structural identity to eukaryote transporters and an N-terminal cytoplasmic proline/serine-rich putative SH3 binding domain. Br J Clin Pract, 1994 May-Jun, 48(3), 130 - 2 Vertebral osteomyelitis in Qatar; Al Soub H et al.; Twenty-eight cases of vertebral osteomyelitis were diagnosed at Hamad General Hospital in Qatar between January 1988 and December 1991: 16 (57.2%) cases were tuberculous spondylitis, 6 (21.4%) Brucella spondylitis, 3 (10.7%) Staphylococcus aureus spondylitis, 2 (7.1%) Salmonella spondylitis, and 1 (3.6%) Pseudomonas cepacia spondylitis . Plain vertebral x-ray films and CT scans were abnormal in all patients . Bone and gallium scans were abnormal in 87.5% and 64% of cases respectively . Clinical manifestations, haematological and radiological investigations were not able to differentiate between the causes . Serological tests were helpful in diagnosing Brucella spondylitis . CT-guided needle biopsy was able to identify the aetiology in 90% of cases . We conclude that invasive tests are still needed to establish the microbiological diagnosis and to guide antimicrobial therapy in most cases of vertebral osteomyelitis. Epidemiol Mikrobiol Imunol, 1994 May, 43(2), 47 - 54 {Salmonellosis in the Czech Republic 1989-1993}; Sramova H et al.; The authors discuss the incidence of salmonelloses in the Czech Republic and analyze changes which developed during the period from 1989 to 1993 . In 1989 a steep rise of cases of salmonellosis was recorded; 34,435 cases were notified which, as compared with the previous period, is a threefold rise of the incidence . This trend persists already for five years . So far the largest number of notified cases (43,558) was recorded in 1992 . The death rate and case fatality rate from salmonellosis has been low for some years: the number of deaths is approximately 20-25 people per year . The salmonellosis epidemic in 1989-1993 spread to all regions of the Czech Republic . The most heavily affected regions are the South Moravian, North Moravian, East Bohemian and West Bohemian region . As to age distribution, it was revealed that the highest values of specific morbidity are in children in their first year of life and in 1-4-year-olds and that the greatest increment in 1989-1993 was notified in 1-4-year-old and in 5-9-year-old children . The dominating aetiological agent was Salmonella enteritidis, probably phagotype 8 . Its ratio in the isolation of Salmonella strains is 87% . The largest number of epidemics in 1989-1991 was notified in communities with a communal catering type (workers canteens, catering for nursery schools and schools) and in communities where half the epidemics developed during family festivities (weddings, graduation, funerals, birthdays, pig-slaughtering feasts etc.) . During the last two years the number of epidemics which develop in workshops producing foods and in private confectionaries is rising.(ABSTRACT TRUNCATED AT 250 WORDS) Yakugaku Zasshi, 1994 May, 114(5), 342 - 50 {Antimutagenic and bactericidal substances in the fruit body of a Basidiomycete Agaricus blazei, Jun-17}; Osaki Y et al.; The fruit body of a Basidiomycete Agaricus blazei, Jun-17 (Himematsutake) was extracted with hexane and chloroform-methanol (2:1, v/v), and the antimutagenic effect of the extracts was examined using an Ames/Salmonella/microsome assay . Both extracts of Agaricus inhibited the mutagenicity of benzo{a}pyrene(B{a}P) . The hexane extract was purified by silica gel column chromatography and high performance liquid chromatography (HPLC), and linoleic acid was isolated as a main substance having antimutagenic activity . Fr . IIa, IIb, IIc and IIb, which reduced the number of His+ revertant colonies induced by B{a}P, were separated from the chloroform-methanol extract by silica gel column chromatography and HPLC . An antimutagenic substance in Fr . IIa was linoleic acid . From Fr . IIb, a bactericidal, not antimutagenic, substance was isolated and identified as 13-hydroxy cis-9, trans-11-octadecadienoic acid (13ZE-LOH) . Antimutagenic substances in Fr . IIc and IId were not purified . The possible source and mechanism of formation of 13ZE-LOH are discussed. Bull Tokyo Dent Coll, 1994 May, 35(2), 67 - 78 Polyclonal B cell activation, endotoxin tolerance, and limulus tests of endotoxin preparations of some periodontopathogens; Inada K et al.; Potencies of polyclonal B-cell activation in C3H/HeN mice of Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis endotoxins were 0.36, 0.13 and 0.04, taking Salmonella abortusequi as 1.0 . F . nucleatum and P . gingivalis endotoxins showed positive reactions in C3H/HeJ mice . Most activities in C3H/HeN other than that of F . nucleatum were suppressed by polymyxin B . In C3H/HeJ mice, similar inhibitions were only 60% for P . gingivalis and hardly observed with F . nucleatum . The resistances to polymyxin B could be due to protein in the endotoxins . A promoting effect of T cells added to B cells was observed only in the activity of F . nucleatum endotoxin in C3H/HeJ mice; there was no influence in other groups . Test endotoxins had nearly the same ability to produce colony stimulating factor as did references and could not produce the factor in tolerant mice . The clinical significance of tolerance is discussed . Regression lines of endotoxin doses and limulus activities of test endotoxins and Salmonella were parallel, either in specific or non-specific tests . The lines of two test groups were also parallel; values obtained by two tests were very close . These data indicate that the test endotoxins did not contain (1-3)-beta-D-glucan and elicited qualitatively similar limulus reactions to that of the reference, despite their different chemical natures . In conclusion, these test preparations had an endotoxicity similar to that of the reference and contribute to produce periodontitis through polyclonal B cell activation. Circ Shock, 1994 May, 43(1), 9 - 17 Priming of phagocytes for reactive oxygen production during hepatic ischemia-reperfusion potentiates the susceptibility for endotoxin-induced liver injury; Liu P et al.; Plasma levels of glutathione disulfide (GSSG) as an indicator of a vascular oxidant stress, tumor necrosis factor-alpha (TNF-alpha) formation, and liver injury (alanine aminotransferase activity, histology) were monitored in male Fischer rats after 30 min of hepatic ischemia followed by up to 4 hr of reperfusion . The injection of 1 mg/kg Salmonella enteritidis endotoxin at 30 min of reflow potentiated the postischemic oxidant stress and liver injury . TNF-alpha levels increased from 10 +/- 7 pg/ml (baseline) to 3,553 +/- 738 pg/ml after ischemia-reperfusion followed by endotoxin, or to 3,670 +/- 508 pg/ml after endotoxin alone . Depletion of serum complement before ischemia attenuated the endotoxin-mediated increase of reactive oxygen formation by 70% but did not affect TNF-alpha levels . Complement activation with cobra venom factor (CVF) during reperfusion had an effect similar to that of endotoxin on the oxidant stress and liver injury . CVF did not increase TNF-alpha formation during reperfusion . Kupffer cells and neutrophils isolated from the postischemic liver 2.5 hr after endotoxin injection generated 600% and 400% more superoxide, respectively, than cells isolated from control livers . The results demonstrate a substantial priming of hepatic phagocytes for reactive oxygen production but not TNF-alpha formation, even after short periods of hepatic ischemia, and the vulnerability of the postischemic liver to severe endotoxin-induced injury . Activated complement seems to be mainly responsible for the effects . These results may explain the high risk for hepatic failure after extensive liver resection and hypovolemic shock. Zh Mikrobiol Epidemiol Immunobiol, 1994 May-Jun, (3), 10 - 4 {The comparative characteristics of preparations of Yersinia pestis capsular antigen F1 obtained from producer strains with different lipopolysaccharide structures}; Gremiakova TA et al.; The main protective antigen of the causative agent of plague is capsular antigen F1 . The preparations of this antigen isolated from Y.pestis strain EV are characterized by a high content of polysaccharide chains of endotoxins . This can be avoided by using R-variants of bacteria as producers . In this work the comparative study of the preparations of antigen F1 obtained from Y.pestis strain EV, Escherichia coli producer strain HB101 pFS1 with the complete structure of LPS and Salmonella minnesota producer strain Re595 pFS1 with maximally reduced LPS has been made . As revealed in this study, the physico-chemical properties of these preparations (the isoelectric point, electrophoretic mobility, the molecular weight of subunits) are identical . The preparation of antigen F1 obtained from S.minnesota has been found to give the highest yield and to have the lowest content of polysaccharide admixtures . This preparation has proved to possess the maximal protective potency, which may be linked with the adjuvant and immunogenic activity of microadmixtures of glycolipid Re, contained in F1. Q J Med, 1994 May, 87(5), 301 - 9 Unusual manifestations of salmonellosis--a surgical problem; Lalitha MK et al.; From January 1981 to December 1992, of 6250 cases of salmonellosis treated at the Christian Medical College and Hospital, Vellore, India, 100 patients with focal pyogenic infection caused by salmonellae required surgical intervention in addition to medical therapy . Thirty-one had involvement of the hepatobiliary system, and 10 more had other intra-abdominal infections . Involvement of bone and joint as well as soft tissue constituted 15% each . The site of infection in patients with soft tissue abscesses included skin (7), parotid (2), thyroid (2), breast (1) inguinal node (1), branchial sinus (1) and injection site (1) . Three patients had arterial infections . Noteworthy among the cases of genital infections was one case of salmonella infection in a pre-existing hydrocele, and one case of epididymo-orchitis with a loculated salmonella infection . Salmonella infection in a pre-existing ovarian cyst was seen in a patient with endometriosis . The salmonella serotypes most frequently encountered were S . typhi (36) and S . typhimurium (36), followed by S . paratyphi A (15) . The importance of recognition of these protean manifestations of salmonellosis in an endemic setting is discussed . The microbiological evaluation of properly obtained specimens is mandatory in such unusual pyogenic infections. Int J Prosthodont, 1994 May-Jun, 7(3), 234 - 8 Immersion disinfection of irreversible hydrocolloid impressions with sodium hypochlorite . Part I: Microbiology; Beyerle MP et al.; Current American Dental Association infection control guidelines recommend immersion disinfection of irreversible hydrocolloid impressions, and this study further defines the parameters for use of sodium hypochlorite . Sodium hypochlorite has been shown to be an effective disinfectant for impressions; however, it has not been fully evaluated for optimum immersion time and concentration . In this study, irreversible hydrocolloid impressions contaminated with different bacteria were immersed in varying concentrations of sodium hypochlorite for 1, 5, or 10 minutes . Dilute solutions of sodium hypochlorite (0.525% or 0.0525%) produced a 4-log10 (99.99%) reduction in colony-forming units of Staphylococcus aureus, Salmonella choleraesuis, or Pseudomonas aeruginosa after 1 to 5 minutes' immersion . Full-strength sodium hypochlorite (5.25%) required 5 minutes to produce a 4-log10 reduction of Bacillus subtilis . A 4-log10 reduction of Mycobacterium bovis was not obtained under any conditions examined. J Clin Microbiol, 1994 May, 32(5), 1135 - 41 Epidemiologic analysis of sporadic Salmonella typhi isolates and those from outbreaks by pulsed-field gel electrophoresis; Thong KL et al.; Pulsed-field gel electrophoresis (PFGE) was used to compare and analyze 158 isolates of Salmonella typhi from five well-defined outbreaks of typhoid fever in Malaysia and also isolates involved in sporadic cases of typhoid fever occurring during the same period . Digestion of chromosomal DNAs from these S . typhi isolates with the restriction endonucleases XbaI (5'-TCTAGA-3'), SpeI (5'-ACTAGT-3'), and AvrII (5'-CCTAGG-3') and then PFGE produced restriction endonuclease analysis (REA) patterns consisting of 11 to 24 DNA fragments ranging in size from 20 to 630 kbp . Analysis of the REA patterns generated by PFGE after digestion with XbaI and SpeI indicated that the S . typhi isolates obtained from sporadic cases of infection were much more heterogeneous (at least 13 different REA patterns were detected; Dice coefficient, between 0.73 and 1.0) than those obtained during outbreaks of typhoid fever . The clonal nature and the close genetic identities of isolates from outbreaks in Alor Setar, Penang, Kota Kinabalu, Johor Bahru, and Kota Bahru were suggested by the fact that only a limited number of REA patterns, which mostly differed by only a single band, were detected (one to four patterns; Dice coefficient, between 0.82 and 1.0), although a different pattern was associated with each of these outbreaks . Comparison of REA patterns with ribotyping for 18 S . typhi isolates involved in sporadic cases of infection showed a good correlation, in that 72% of the isolates were in the same group . There was no clear correlation of phage types with a specific REA pattern . We conclude that PFGE of s . typhi chromosomal DNA digested with infrequently cutting restriction endonucleases is a useful method for comparing and differentiating S . typhi isolates for epidemiological purposes. Shock, 1994 May, 1(5), 362 - 5 Plasma endotoxin concentration after an intraperitoneal injection of endotoxin in fed and fasted suckling rats; Yoshioka T et al.; In the adult host response to endotoxin (lipopolysaccharide (LPS)) is dose-related . An intraperitoneal injection is commonly used for LPS administration in small animals . However, plasma endotoxin concentration following an intraperitoneal bolus injection of LPS is not well known . This study was performed to evaluate plasma endotoxin concentration following a bolus intraperitoneal injection of LPS in both fed and 24 h fasted 10 day old rats . Plasma endotoxin concentration increased in a dose-dependent manner after LPS injection (.03 or .1 mg/kg Salmonella enteritidis LPS) in both fed and fasted rats . Plasma endotoxin concentrations were higher (p < .05) in fed than fasted rats . A high dose of LPS (.1 mg/kg) induced 95 and 40% mortality in fed and fasted rats, respectively . A low dose of LPS (.03 mg/kg) induced 26.7% mortality in fed rats but no mortality in fasted rats . The hematocrit was significantly lower in fed than fasted rats . Plasma endotoxin inactivation was similar in fed and fasted rats . Host response appears to be related to plasma endotoxin concentration. Int J Food Microbiol, 1994 May, 22(2-3), 97 - 103 Usefulness of molecular genetic markers in the typing of Salmonella enterica serovar Enteritidis causing a food-borne outbreak; Gonzalez-Hevia MA et al.; A combination of serotyping-phagetyping and three molecular genetic markers (plasmid analysis, chromosomic DNA restriction pattern and ribosomal RNA gene restriction pattern or ribotyping) was used in the typing of Salmonella enterica causing a food-borne outbreak . The isolates analysed, 29 from stools and eight from foods, belonging to serovar Enteritidis-phagetype A, carried a 36-MDa plasmid, showed a similar DNA restriction pattern and the same ribopattern . These data indicate that only one strain was involved . The DNA pattern and ribopattern of this strain were indistinguishable from the patterns of a serovar Enteritidis-phagetype A strain which has caused salmonellosis in Asturias, Spain, since, at least, 1984. Infect Immun, 1994 May, 62(5), 1520 - 7 CD14 and CD11b mediate serum-independent binding to human monocytes of an acylpolygalactoside isolated from Klebsiella pneumoniae; Hmama Z et al.; A water-soluble acylpolygalactosyl (APG) of 34 kDa was obtained from the Klebsiella pneumoniae membrane by alkaline hydrolysis and delipidation . APG comprises a poly(1,3)galactose chain, a core, and a lipid moiety made of a glucosamine disaccharide with two N-linked beta OH-myristates . The monocyte binding sites for APG were investigated by flow cytometry . Biotin-labelled APG (Biot-APG) bound to monocytes at 4 degrees C in the absence of serum, calcium, and magnesium . The binding was dose dependent, saturable, and displaced by unlabelled APG . Neither the polysaccharide chain present in APG-related molecules nor the PPi group or additional ester-linked myristates and palmitates were required for APG binding . The role of CD11b and CD14 was demonstrated by competitive inhibition with monoclonal antibodies and by the uptake of APG by these solubilized proteins . APG was rapidly internalized into monocytes at 37 degrees C while CD14 and CD11b/CD18 molecules were partially down-modulated . Lipopolysaccharides (LPS) from the same K . pneumoniae strain and from Escherichia coli and Salmonella minnesota partially competed for Biot-APG binding in the absence but not in the presence of serum . When altered by alkaline hydrolysis, those LPS became strong competitors for APG binding . It was concluded that alkaline hydrolysis of the K . pneumoniae membrane yielded molecules structurally related to LPS which bind to LPS membrane receptors in the absence of serum. J Biol Chem, 1994 Apr 8, 269(14), 10675 - 82 Intermediate filament-like network formed in vitro by a bacterial coiled coil protein; Hurme R et al.; The TlpA protein encoded by the virulence plasmid of Salmonella enterica is an alpha-helical 371-amino acid protein possessing characteristics similar to eukaryotic coiled coil proteins (Koski, P., Saarilahti, H., Sukupolvi, S., Taira, S., Rikkonen, P., Osterlund, K., Hurme, R., and Rhen, M . (1992) J . Biol . Chem . 267, 12258-12265) . In this paper we have investigated inter- and intramolecular associations and the morphology of structures formed by TlpA . Dynamics and temperature stability of TlpA dimers were studied by examining the feasibility and conditions in which TlpA would form an artificial heterodimer with its truncated derivative . Formation of heterodimers, bridged by Cu(2+)-catalyzed air oxidation of adjacent Cys residues, showed that TlpA dimers are dynamic chain exchanging structures at 37 degrees C, whereas they were nonexchanging at room temperature or on ice . Chemical cross-linking suggested higher order interaction between TlpA dimers . Electron microscopy studies revealed two levels of TlpA organization in vitro: thin filaments and rods, 2-5 nm in diameter, and a higher ordered filament network consisting of tonofilament-like formations with a diameter of 8-15 nm . Electron microscopy of thin-sectioned Escherichia coli over-producing TlpA showed an extraordinary intracellular assembly of proteinacious lamellae with a striated appearance and a 38-nm periodicity . This study describes for the first time a bacterial protein capable of organizing itself into an ordered and suspectedly dynamic intermediate filament-like architecture. Tierarztl Prax, 1994 Apr, 22(2), 141 - 5 {Function and development of autochthonous intestinal flora in domestic poultry}; Gerlach H; A report is given on the development and function of the autochthonous intestinal flora in poultry . The relevance of this flora for infections with Salmonella spp., both following egg transmission and oral intake from the 4th to 6th week of life is discussed. J Trop Med Hyg, 1994 Apr, 97(2), 87 - 90 Childhood bacterial diarrhoea in a regional hospital in Saudi Arabia: clinico-aetiological features; al-Jurayyan NA et al.; Over a one-year period, 210 paediatric patients, who were admitted with acute diarrhoea to a regional hospital in the south-western region of Saudi Arabia, were retrospectively reviewed for bacterial enteropathogens . Bacterial pathogens were isolated from 66 (31.4%) patients, with Shigella being the most common (17.1%), followed by Salmonella (10.5%), and enteropathogenic Escherichia coli (EPEC) (3.8%) . Major clinical findings associated with bacterial diarrhoea are similar to those reported before . Our results suggest that bacterial pathogens constitute a major cause of acute childhood diarrhoea in hospitalized children in Al-Baha province . Further prospective community based studies are needed to identify the pattern and risk factors of acute childhood diarrhoea in the region. Arch Biochem Biophys, 1994 Apr, 310(1), 89 - 96 Generation of one set of murine monoclonal antibodies specific for globo-series glycolipids: evidence for differential distribution of the glycolipids in rat small intestine; Kotani M et al.; We generated four murine monoclonal antibodies (MAbs) specific for globo-series glycolipids by immunizing C3H/HeN mice with these purified glycolipids adsorbed to Salmonella minnesota followed by fusion with mouse myeloma cells . By use of a wide variety of glycolipids, the precise structures recognized by these MAbs were elucidated through an enzyme-linked immunosorbent assay and an immunostaining on thin-layer chromatography . These four MAbs, designated as BGR23, BGR47, BMR26, and BGR27, exhibited highly restricted binding specificities, reacting only with the globo-series glycolipids Gb3Cer, III3Gal alpha-Gb3Cer, Gb4Cer, and IV3GalNAc alpha-Gb4Cer, respectively, which were used for immunization . None of the other various glycolipids or gangliosides were recognized . We determined the localization of these globo-series glycolipids in adult rat small intestine by means of an immunofluorescence technique with these MAbs . Our study revealed that there is a differential distribution of these glycolipids in the rat tissue . III3Gal alpha-Gb3Cer was demonstrated on the cryptic cells and circular muscle, whereas Gb4Cer was localized on both the circular and longitudinal muscles . The expression of Gb3Cer was associated with the epithelium and the capillary endothelial cells in the lamina propria mucosae as well as with the tunica submucosa, whereas IV3GalNAc alpha-Gb4Cer was detected on the epithelium, capillary endothelial cells in the lamina propria mucosae, and both the muscle layers. Epidemiol Infect, 1994 Apr, 112(2), 253 - 61 Insertion sequence IS200 fingerprinting of Salmonella typhi: an assessment of epidemiological applicability; Threlfall EJ et al.; When Pst I-generated digests of genomic DNA from each of the type strains of 49 of the Vi phage types of Salmonella typhi were probed with a PCR-amplified IS200 gene probe, all strains were found to possess at least 11 IS200 elements carried on fragments in the range 24.2-1.2 kb . Fourteen fingerprints were identified but two patterns designated IS200Sty1 and IS200Sty2 predominated . In one strain, a plasmid-mediated IS200 element was identified . When IS200 fingerprinting was applied to epidemiologically-unrelated strains of S . typhi isolated in Ecuador, 3 patterns were identified in 10 strains belonging to 9 different phage types . It is concluded that Vi phage typing remains the method of choice for the primary differentiation of S . typhi but that IS200 fingerprinting may be of limited use in laboratories which do not have access to phage typing. J Biol Chem, 1994 Apr 1, 269(13), 9533 - 8 Selection of antibody single-chain variable fragments with improved carbohydrate binding by phage display; Deng SJ et al.; A single-chain variable fragment (Fv) version of a murine monoclonal antibody, Se155-4, specific for Salmonella serogroup B O-polysaccharide, was used as a model system for testing monovalent phage display as a route for enhancing the relatively low affinities that typify anti-carbohydrate antibodies . Random single-chain Fv mutant libraries generated by chemical and error-prone polymerase chain reaction methods were panned against the serogroup B lipopolysaccharide . Panning of a randomly mutated heavy chain variable domain library indicated selection for improved serogroup B binders and yielded six mutants, five of which showed wild type activity by enzyme immunoassay . Two of these were apparently selected on the basis of better functional single-chain Fv yield in Escherichia coli . A heavy chain mutation (Ile77-->Thr) in one mutant, 3B1, appeared to have a particularly dramatic effect, resulting in yields of approximately 120 mg/liter of functional periplasmic product . The sixth mutant, 4B2, had complementarity determining region 1 (CDR1) and CDR2 mutations and demonstrated 10-fold improved binding, by enzyme immunoassay, relative to the wild type . Extensive analysis of antigen-antibody interactions indicated that the improved binding properties of 4B2 were attributable to a higher association rate constant and interaction with an epitope that is larger than the trisaccharide recognized by the wild type . None of the mutations involved known trisaccharide contact residues; this was consistent with analysis of wild type and mutant single-chain Fvs by titration microcalorimetry . Examination of the structure indicated that two mutations in the heavy chain CDR2 provided improved surface complementarity between the protein and the extended epitope encompassing 2 additional hexose residues . However, introduction of only the CDR2 mutations into the wild type structure failed to confer the improved binding properties of 4B2, indicating an indirect effect by the more distant mutations . Panning of randomly mutated light chain variable domain and full-length single-chain Fv mutant libraries did not yield mutants with improved assembly or binding properties. Cancer Res, 1994 Apr 1, 54(7 Suppl), 1991s - 1993s Suppression of hydroperoxide-induced cytotoxicity by polyphenols; Nakayama T; A variety of synthetic and dietary polyphenols protect mammalian and bacterial cells from cytotoxicity induced by hydroperoxides, especially hydrogen peroxide (H2O2) . Cytotoxicity of H2O2 on Chinese hamster V79 cells was assessed with a colony formation assay . Cytotoxicity and mutagenicity of H2O2 on Salmonella TA104 were assessed with the Ames test . SOS response induced by H2O2 was investigated in the SOS chromotest with Escherichia coli PQ37 . The polyphenol-bearing o-dihydroxy (catechol) structure, i.e., nordihydroguaiaretic acid, caffeic acid ester, gallic acid ester, quercetin, and catechin, were effective for suppression of H2O2-induced cytotoxicity in these assay systems . In contrast, neither ferulic acid ester-bearing o-methoxyphenol structure nor alpha-tocopherol were effective, indicating that o-dihydroxy or its equivalent structure in flavonoids is essential for the protection . There are many reports describing that polyphenols act as prooxidants in the presence of metal ions . Our results suggest, however, that they act as antioxidants in the cells, when no metal ions are added to the medium. J Infect Dis, 1994 Apr, 169(4), 927 - 31 Salmonella typhi vaccine strain CVD 908 expressing the circumsporozoite protein of Plasmodium falciparum: strain construction and safety and immunogenicity in humans; Gonzalez C et al.; rcsp, encoding amino acids 21-398 of Plasmodium falciparum circumsporozoite protein (CSP), under control of tacP was integrated into the chromosomal delta aroC locus of attenuated delta aroC, delta aroD Salmonella typhi CVD 908 . By immunoblot and ELISA, rCSP expression was greater from a multicopy plasmid than from the single chromosomal gene . CVD 908 omega (delta aroC1019::tacP-rcsp) was well tolerated by 10 volunteers who were fed two doses of 5 x 10(7) organisms 8 days apart . Seven subjects excreted the vaccine strain for 1-3 days . All subjects developed serologic responses to O and H antigens of the live vector, whereas 3 vaccinees responded to the foreign antigen: 1 developed an 80-fold rise in serum anti-sporozoite antibody, another had a 4-fold rise in antibody to a recombinant portion of CSP (residues 309-345), while a third vaccinee developed CSP-specific CD8+ cytotoxic T lymphocyte activity . This is the first report of attenuated S . typhi eliciting a human serologic or a cytotoxic T lymphocyte response to a foreign protein . Improved foreign gene expression should enhance immunogenicity. Indian J Med Sci, 1994 Apr, 48(4), 85 - 8 Multidrug resistant Salmonella typhi in Bangalore, south India; Rathish KC et al.; A total of 204 strains of salmonella were isolated in blood cultures during the year 1991 outbreak of enteric fever in and around Bangalore . Out of this, 190 were S . typhi, 6 S . paratyphi A, 5 S . typhimurium and 3 S . choloraesuis . Antibiogram of 190 strains of S . typhi showed resistance of 94.7%, 95.8% and 96.9% to chloramphenicol, ampicillin and cotrimoxazole and sensitivity of 65.3%, 88.4% and 94.2% to gentamycin, norfloxacin and ciprofloxacin respectively . Minimum inhibitory concentrations (MIC) of chloramphenicol were between 360 mcg and 640 mcg per ml . There was high degree (94.7%) of triple drug resistance to chloramphenicol, ampicillin and cotrimoxazole. Wei Sheng Wu Xue Bao, 1994 Apr, 34(2), 152 - 5 {A new serotype of Salmonella III b}; Su F et al.; A new serotype of Salmonella No . S . 3337 was isolated from the intestinal content of reptile a snake in August 1989 . Providing with Salmonella biological characteristics, it could be classified into subspecies III b because it utilized sodium malonate, attacked lactose promptly, ONPG positive, and did not ferment dulcitol . H antigens appeared diphasic . Antigenic analysis shows that it represents a new serotype with an antigenic formula of 65 : z : z55. Eur J Clin Microbiol Infect Dis, 1994 Apr, 13(4), 307 - 10 Eradication of convalescent-phase Salmonella carriage in children with two oral doses of pefloxacin; Raymond J et al.; Fifteen children (age range 1.5 months to 7.2 years), who were excluded from schools or nurseries due to asymptomatic convalescent-phase non-typhoidal Salmonella carriage, received two oral doses of pefloxacin (12 mg/kg on days 1 and 4) and were examined on days 10, 30, 45 and 60 . Definitive eradication was observed in 13 patients, all of whom had initial low Salmonella counts in stools and were culture-negative by day 10 . In the two patients who failed to respond, the same treatment was effective when repeated 4 and 6 months later respectively . No side-effects were observed . In six other children, considered as controls, eradication by day 10 was observed in only one case after administration of amoxicillin for eight days . Two oral doses of pefloxacin could be a useful and safe means for eliminating Salmonella carriage in young children. Int J Food Microbiol, 1994 Apr, 22(1), 1 - 9 Competitive exclusion of Salmonella enteritidis in chicks by treatment with a single culture plus dietary lactose; Behling RG et al.; Oral inoculation of lactose utilizing cecal bacteria plus 2.5% lactose treatments were tested in young chicks for protective efficacy against infection by Salmonella enteritidis . One-day-old chicks were treated with cecal bacteria upon arrival and challenged orally on day 3 with 10(4)-10(6) cfu S . enteritidis . A single culture identified as Escherichia coli O75:H10 was found significantly more protective than all other isolates tested . This isolate excreted a metabolite(s) in vitro that was inhibitory towards the growth of S . enteritidis . The results of this study indicate that discovery of protective strains can be facilitated by screening isolates in vitro for lactose utilization and growth inhibition of S . enteritidis before administration of treatment. Anal Biochem, 1994 Apr, 218(1), 63 - 73 Detailed structural characterization of lipid A: electrospray ionization coupled with tandem mass spectrometry; Chan S et al.; Previous studies have defined specific functional relationships within monophosphoryl lipid A (MLA) preparations . To extend this understanding to all contributing entities, MLA samples have been structurally characterized using electrospray ionization, collision-induced dissociation (CID), and tandem mass spectrometry (MS/MS) . MLA profiles of Salmonella minnesota Re595 have been compared with Shigella flexneri for sample type and component distribution . In excess of 20 individual structures compose each sample, which differ considerably in abundance but little in composition . Component heterogeneity can be directly related to alkane chain length, "lipid X"-type analogs, and variations in esterification to the core 2-amino-2-deoxydisaccharide, GlcNH2 beta(1-6)GlcNH2 . The previously defined heptaacyl structure in S . minnesota Re595 was identified at only 15% with the most abundant species a hexaacyl analog . Profiles of S . flexneri MLA show an absence of any heptaacyl analog, with the pentaacyl component the most abundant . To confirm structural relationships, ions from both samples were fragmented by CID and separated by MS/MS . Product ion spectra proved to be identical, showing a series of acyl losses and glycosidic cleavage fragments . Since the position of each acyl group has been previously established in S . minnesota Re595 MLA, ions in the CID spectrum of S . flexneri sample could be structurally assigned . Knowledge of these structural details, their isolation, and biological testing may provide components of unique immune adjuvancy or block-selective deleterious endotoxic processes. J Physiol, 1994 Apr 1, 476(1), 177 - 86 Interleukin-1 beta production in the rabbit brain during endotoxin-induced fever; Nakamori T et al.; Interleukin-1 beta (IL-1 beta) production in the brain and the spleen was investigated in rabbits made febrile by intravenous (I.V.) injection of endotoxin, or human recombinant IL-1 beta (hIL-1 beta) . The endotoxin used in the present study was the lipopolysaccharide (LPS) of Salmonella typhosa endotoxin . Monophasic fever was induced by I.V . injection of a low dose of LPS (0.02 micrograms kg-1) and biphasic fever by I.V . injection of a large dose of LPS (4 micrograms kg-1), a sublethal dose of LPS (40 micrograms kg-1) or hIL-1 beta (2 micrograms kg-1) . In situ hybridization and immunohistochemical studies revealed that, although no IL-1 beta production was observed in the brain at 1 and 3 h after injection of a low dose of LPS (0.02 micrograms kg-1) or of hIL-1 beta (2 micrograms kg-1), IL-1 beta production was demonstrated in organum vasculosum laminae terminalis (OVLT) and some cells around the blood vessels in the parenchyma 1 h after 4 micrograms kg-1 LPS . IL-1 beta production was detected throughout the brain after 40 micrograms kg-1 LPS . Pretreatment with indomethacin, an inhibitor of prostaglandin synthesis, did not affect IL-1 beta production in the brain induced by 4 micrograms kg-1 LPS . The cell type which produces IL-1 beta in the OVLT following LPS injection was confirmed to be a macrophage by electron microscopy . The cells producing IL-1 beta in the parenchyma were determined to be microglial cells . In the spleen, each dose of LPS induced a significant increase in IL-1 beta production in polymorphonuclear cells and macrophages in the red pulp 1 h after injection . However, 2 micrograms kg-1 hIL-1 beta did not induce IL-1 beta production in the spleen . The present results show clearly that systemic administration of LPS induces IL-1 beta production in the OVLT which may be responsible for induction of the second phase of biphasic fever . The production of IL-1 beta in the OVLT was not attributable to the action of peripherally synthesized IL-1 beta or prostaglandins. An Med Interna, 1994 Apr, 11(4), 173 - 6 {Pyogenic liver abscess . Study of 20 patients treated with percutaneous drainage}; de Miguel J et al.; We conducted a descriptive study of pyogenous hepatic abscesses (PHA) and their treatment with percutaneous drainage and antibiotherapy in the general hospital of Galdacano between 1989 and 1992 . We assessed prevalence, clinical characteristics, responses to treatment, evolution and complications . We studied 20 PHAs in adults confirmed through puncture guided with echography and/or computerized tomography . We considered as causal germs those isolated in the abscess and/or hemocultures . All the patients were treated with catheter drainage and antibiotics . After discharge, follow-up and regular TC controls were performed at least for 6 months . The average age of the patients was 56 +/- 3 years and the men/women rate was 2.5:1 . The most frequent origin of the infection was cholecystitis/cholangitis in 50% of patients and hydatidic cysts in 20% . Twelve patients had isolated abscess and 8 patients, multiple abscesses . The diagnostic sensitivity was 95% for the echography and 100% for CAT . The most frequent germs were E . Coli, Streptococcus, K . pneumoniae and Salmonella spp . In three cases, it was not possible to bacteriologically identified the germ . After drainage, the abscesses disappeared in 16 patients . The average duration of percutaneous drainage was 10 days . Three patients required surgery after drainage due to complications or incomplete drainage; two other patients required extirpation of hydatidic cysts . The mortality rate was 10%, although it was not related to PHA . We did not observe any differences between isolated or multiple abscesses with regard to prognosis . The drainage guided by echography and/or CT, associated to antibiotic therapy, is a successful technique for the treatment of PHAs in most patients.(ABSTRACT TRUNCATED AT 250 WORDS) Enferm Infecc Microbiol Clin, 1994 Apr, 12(4), 197 - 9 {Subtyping of Salmonella enteritidis using a new phage typing protocol}; Alonso R et al.; BACKGROUND: Salmonella enteritidis isolates have increased since 1982 from 20% to more than 70% in 1992 in Spain . Currently available phenotypic subtyping methods have a low discriminatory power . In order to improve the phage-typing discriminatory power of Alonso's method, new bacteriophages have been developed . METHODS: Wild bacteriophages were isolated from different waste waters . Lysogenic bacteriophages were induced with mitomycin C . RESULTS: We selected 7 wild and 2 lysogenic bacteriophages, considering their stability, reproducibility and discriminatory power . Ten phage-types were also described . CONCLUSIONS: Using Simpson's index of diversity, the proposed scheme is more discriminatory power than Alonso's previously described one . It is necessary to compare it with other subtyping methods, like new molecular methods, before establishing its usefulness as a complementary marker. Enferm Infecc Microbiol Clin, 1994 Apr, 12(4), 187 - 92 {Microbial drug resistance and the presence of plasmids in Salmonella strains isolated from different sources}; Luque A et al.; BACKGROUND: To establish the relationship between the presence of plasmid and their antimicrobial resistance of Salmonella strains . METHODS: We tested 171 strains of Salmonella isolated from different sources: natural waters (73 strains), food (23 strains), and from clinical samples (75 strains) . The disk diffusion method was used to test the antimicrobial susceptibility of the strains to 13 drugs . Plasmid analysis were performed by agarose gel electrophoresis technique . RESULTS: Antimicrobial resistances of the strains significantly varies according to their primary isolation sources . Strains isolated from the water environment exhibited a full susceptibility to cephalothin and colistin, whilst all the strains isolated from food were sensitive to ampicillin, gentamicin, kanamycin, neomycin, tobramycin and trimethoprim-sulphamethoxazole . On the other hand, resistances to colistin, gentamicin and trimethoprim-sulphamethoxazole were not found in clinical isolates . From the 171 Salmonella strains tested, only 12.2% were sensitive to all the antimicrobials . The most frequently antibiotic resistances detected were to streptomycin (49.3%), tetracycline (33.1%) and nalidixic acid (30.7%) . The percentage of strains that harboured plasmids was different depending on the source of isolation, ranging from 41.4% for water isolated, 76% for clinical isolates and 86.9% for food isolates . The relationship between antimicrobial resistance and plasmid presence is very close, since higher percentages of resistance to chloramphenicol, carbenicillin, cephalothin, kanamycin, neomycin, nalidixic acid and trimethoprim-sulphamethoxazole were obtained in strains containing plasmids . CONCLUSIONS: (a) The most frequently resistance detected in strains of Salmonella was to streptomycin (49.3% of the strains) . On the other hand, only 0.6% of the strains were resistant to gentamicin . (b) Percentages of resistance to some antibiotics was higher in strains harbouring plasmids, that implies a relationship between the plasmid presence and the antibiotic resistance in Salmonella . (c) Curing of extrachromosomic elements by acridine orange showed a percentage of resistance lost greater than 70% for cephalothin, kanamycin, neomycin, and nalidixic acid . This indicates that the resistance to those antibiotics is mainly linked to plasmids . In the case of the unusual nalidixic acid-resistance, previously described in Shigella strains, suggests that are needed more studies to demonstrate the direct association between antimicrobial resistance and presence of plasmids. Gesundheitswesen, 1994 Apr, 56(4), 211 - 4 {Outbreak of Salmonella paratyphi B infections in connection with consumption of smoked fish}; Kuhn H et al.; In the period from 2-10 August 1991 an outbreak caused by S . paratyphi B occurred in five rural areas of the district of Leipzig . Eleven patients and one excreter were involved, and mild forms of disease were observed in most cases . In three of the patients a mixed infection with S . litchfield was diagnosed . Moreover at the same time 21 cases of enteritis caused by S . litchfield and three excreters associated with this serovar were registered in these five rural areas . Both infections with S . paratyphi B and S . litchfield occurred 1-3 days after consumption of smoked halibut . The fish smoked in a smokehouse in the Grimma rural area was delivered to the shops every day . The inspection of the smokehouse and the dispatch department provided no signs of contaminations . In the smoked fish samples investigated subsequently, Salmonella were no longer detectable . The results of typing confirm the identity of all S . paratyphi B strains isolated . The possibilities of contamination of the smoked halibut are discussed . This study emphasises the possible transmission of salmonella by way of the food fish . On that occasion also a non-frequent serovar such as S . paratyphi B can be isolated and a light course of disease as e.g . febrile gastroenteritis can be observed and may attain epidemiological significance. Zentralbl Veterinarmed B, 1994 Apr, 41(2), 113 - 25 Lymphocyte subpopulations in jejunal and ileal Peyer's patches of calves with experimental Salmonella dublin infection; Liebler EM et al.; Changes of lymphocyte subpopulations in jejunal and ileal Peyer's patches (JPP and IPP) of six calves inoculated with Salmonella dublin were investigated at 9 hours, 1, 2, 3 and 7 days post inoculation (p.i.) using immunohistochemistry . Reactive areas and area percentages of B-lymphocytes, as well as CD4+, CD8+ and gamma delta T-lymphocytes within the different compartments of PP were estimated using computer-assisted morphometric analysis . A significant, linear decline of the areas of lymphoid follicles and domes in JPP and IPP due to depletion of B-lymphocytes was found . The rate of decline was similar in JPP and IPP, but more severe in lymphoid follicles than in domes . Intraepithelial cells in follicle-associated epithelium changed from predominantly B-lymphocytes in controls to CD8+ T-lymphocytes in inoculated calves and clusters of B-lymphocytes were observed above domes at days 1 and 2 p.i . Areas of CD4+ and CD8+ T-lymphocytes within lymphoid follicles and domes were increased at 3 and 7 days . p.i . resulting in decreased compartmentalization of the normally segregated T- and B-lymphocyte populations . The increase of CD4+ and CD8+ T-lymphocytes was, however, significant in lymphoid follicles in the JPP only . No significant changes in the amount and distribution of gamma delta T-lymphocytes were observed. Avian Dis, 1994 Apr-Jun, 38(2), 329 - 33 Effect of prolonged administration of dietary capsaicin on broiler growth and Salmonella enteritidis susceptibility; McElroy AP et al.; The effect of continuous (42 days) dietary administration of 5 or 20 ppm capsaicin to broiler chickens on Salmonella enteritidis susceptibility, body weight, and feed efficiency was investigated . Chickens were weighed at 1, 21, and 42 days of age . No significant differences in body weight or feed efficiency were observed . Chickens were challenged with 1 x 10(8) colony-forming units of S . enteritidis at 21, 28, or 42 days of age . The S . enteritidispositive culture rate for cecal tonsils was significantly lower (P < 0.05) in the treatment groups receiving 5 ppm or 20 ppm dietary capsaicin than in the untreated control group at all challenge times . Dietary capsaicin (5 and 20 ppm) resulted in protection against S . enteritidis organ invasion at 28 days in one experiment and at both 21 and 42 days in the other . These results indicate that continual dietary capsaicin administration increases resistance to S . enteritidis colonization and organ invasion throughout the normal growth period without detrimental effects on growth in broiler chickens. Avian Dis, 1994 Apr-Jun, 38(2), 297 - 303 Competitive exclusion of Salmonella enteritidis in Leghorn chicks: comparison of treatment by crop gavage, drinking water, spray, or lyophilized alginate beads; Corrier DE et al.; The protective effect of cecal bacteria cultures on Salmonella enteritidis cecal colonization was evaluated . Competitive-exclusion cultures were administered by crop gavage, in first drinking water, by whole body spray, or encapsulated in alginate beads and provided in feed pans . Leghorn chicks were treated with cultures of cecal bacteria on the day of hatch and challenged orally with 10(4) S . enteritidis 2 days after treatment . Salmonella cecal colonization was evaluated 7 days after challenge . No Salmonella organisms were detected in the ceca of chicks treated with cecal cultures by crop gavage . Chicks treated with cecal cultures in the drinking water or by spray application showed comparable protection and significant decreases (P < 0.05) in the number of Salmonella in the cecal contents compared with untreated controls . The consumption of cecal bacteria encapsulated in alginate beads significantly decreased (P < 0.05) Salmonella cecal colonization compared with control treatment, but it provided less protection than the other treatment methods evaluated. Avian Dis, 1994 Apr-Jun, 38(2), 293 - 6 Evaluation of possible alternatives to double-strength skim milk used to saturate drag swabs for Salmonella detection; Opara OO et al.; The drag-swab Salmonella screening technique was evaluated using less expensive alternatives to double-strength skim milk (2 x SM) as a saturating medium for drag swabs . Ten pre-determined Salmonella-positive poultry houses were studied . In the first phase, Salmonella screening efficiency of drag swabs impregnated with 2 x SM and commercially available canned Carnation evaporated skim milk (CESM) were compared . Results showed CESM to be a less efficient alternative . In the second phase of the study, the Salmonella screening efficiency of drag swabs impregnated with 2% buffered peptone water (BPW), physiological saline (PS), and distilled water (DW) were evaluated along with an unimpregnated drag swab (dry drag swab) (DD) as possible alternatives to 2 x SM . The efficiency of Salmonella detection using various impregnation treatments were in the following order: 2 x SM > PS > BPW > DW > DD. Avian Dis, 1994 Apr-Jun, 38(2), 282 - 8 Horizontal transmission of Salmonella enteritidis and effect of stress on shedding in laying hens; Nakamura M et al.; Horizontal transmission of Salmonella enteritidis in laying hens and the short-term effect of stress on shedding were examined in 32 seven-month-old laying hens . Half were inoculated with 10(5) colony-forming units of S . enteritidis phage type 4, and the remaining half were left uninoculated to study horizontal transmission . Isolation of S . enteritidis from cecal droppings of all hens was attempted every morning . Uninoculated hens rapidly became infected through contaminated drinking water . Introduction of young chickens to the same rearing room and withdrawal of water and feed for 2 days coincided with a rapid increase in the shedding rate of S . enteritidis for a short period of time . The results showed that a short-term increase in the shedding rate of S . enteritidis is associated with short-term exposure to environmental stress. Avian Dis, 1994 Apr-Jun, 38(2), 256 - 61 Effect of selected antibiotics and anticoccidials on Salmonella enteritidis cecal colonization and organ invasion in Leghorn chicks; Manning JG et al.; One-day-old leghorn chicks were placed in floor pens on previously used poultry litter (potentially providing exposure to normal chicken enteric flora) for 7 days and provided feed containing one of several antibiotics or anticoccidials . On day 7, all groups were challenged orally with an isolate of Salmonella enteritidis (10(6) colony-forming units) that was resistant to bacitracin, novobiocin, nalidixic acid, and nitrofurazone . All chicks were killed on day 13, and liver, spleen, and cecal tonsils were cultured . Dietary administration of novobiocin (0.385 g/kg) caused a significant increase (P < 0.05) in positive chick colonization rate (either liver and spleen or cecal tonsils) compared with the unmedicated controls . Similarly, chicks administered dietary nitrofurazone (0.3 g/kg) were infected with S . enteritidis at a significantly greater frequency than the unmedicated controls . A significant decrease in cecal volatile fatty acid concentration, previously shown to influence susceptibility to selected enteric pathogens, was observed in the novobiocin- and nitrofurazone-treated groups . Treatment with chlortetracycline (11.4 g/kg), monensin (0.91 g/kg), or nicarbazin (0.49 g/kg) had no effect on S . enteritidis invasion or colonization . Bacitracin (0.49 g/kg) significantly increased S . enteritidis cecal colonization rate when administered continuously throughout the study . These data support and extend previous investigations involving other salmonellae and indicate that selected antibiotics may increase the severity and frequency of S . enteritidis colonization and invasion rate in leghorn chicks. West Afr J Med, 1994 Apr-Jun, 13(2), 113 - 5 The microflora of bile in Ghanaians; Darko R et al.; Bile was obtained from the gall bladder of 104 patients undergoing cholecystectomy for gall stone disease . Bile was also obtained from the common bile duct and T-tube of 17 patients who also had exploration of common bile duct . During the same period, 148 cholecystectomies were performed . The specimens were sent for culture and sensitivity and 32.7% of the specimens grew bacteria . The factors that were associated with positive culture were emergency cholecystectomy for acute cholecystitis and empyema of gall bladder, carcinoma of gall bladder and obstructive jaundice . The commonest organisms were E . Coli (28.2%) and Klebsiella (17.9%) . Pseudomonas surprisingly formed 10.2% of the cultured organisms . Salmonella that causes typhoid, which is an endemic disease in Ghana, formed only 7.7% of the isolates . Most of the organisms were resistant to Ampicillin and tetracyclines . The antibiotics that most were sensitive to were Gentamicin and Cefuroxime . Therefore the antibiotics that are recommended for use as prophylaxis in biliary tract surgery are Gentamicin or Cefuroxime. Shock, 1994 Apr, 1(4), 254 - 66 CT-1501R selectively inhibits induced inflammatory monokines in human whole blood ex vivo; Rice GC et al.; The effect of (R)-1-(5-hydroxyhexyl)-3,7-dimethylxanthine (CT-1501R; the nonproprietary name for CT-1501R approved by the United States Name Council is lisofylline), an inhibitor of second messenger signaling through phosphatidic acid, on release of endogenous mediators important in the systemic inflammatory response syndrome (SIRS) was studied using the human whole blood ex vivo assay system . Human blood was stimulated with various endotoxin preparations, zymosan, or protein A, and the levels of secreted monokines were measured by enzyme-linked immunosorbent assay . CT-1501R inhibited tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta), and IL-6 release in a dose-dependent manner and was active with all stimuli tested including Salmonella and Escherichia coli-derived endotoxin, endotoxin from both rough and smooth E . coli strains, as well as zymosan and protein A . CT-1501R inhibited monokine release by approximately 50% at 200 microM and 30% at 50 microM and was independent of the relative potency of stimulus . CT-1501R also inhibited IL-1 alpha or IL-1 beta induction of either TNF-alpha or IL-1 beta and inhibited the synergistic effects of stimulation with both human IL-1 beta and murine TNF-alpha on release of human TNF-alpha . Inhibition of monokine release following stimulation with monokine(s) was, in general, greater than that achieved with lipopolysaccharide (LPS) stimulation . Northern blot analysis showed decreased mRNA accumulation of TNF-alpha and IL-1 beta in CT-1501R-treated samples following LPS stimulation suggesting that CT-1501R acts at least in part, at the pretranslational level . In contrast, CT-1501R does not inhibit LPS-stimulated IL-8 or IL-1 receptor antagonist (IL-1ra) release in human whole blood or IL-1 alpha-induced release of PGE2 in human foreskin fibroblast cells . These data suggest that CT-1501R may be of use for clinical intervention in SIRS. J Bacteriol, 1994 Apr, 176(8), 2406 - 14 Hypervariable region IV of Salmonella gene fliCd encodes a dominant surface epitope and a stabilizing factor for functional flagella; He XS et al.; To identify the major antigenic determinant of native Salmonella flagella of antigenic type d, we constructed a series of mutated fliCd genes with deletions and amino acid alterations in hypervariable region IV and in region of putative epitopes as suggested by epitope mapping with synthetic octameric peptides (T.M . Joys and F . Schodel, Infect . Immun . 59:3330-3332, 1991) . The expressed product of most of the mutant genes, with deletions of up to 92 amino acids in region IV, assembled into functional flagella and conferred motility on flagellin-deficient hosts . Serological analysis of these flagella with different anti-d antibodies revealed that the peptide sequence centered at amino acids 229 to 230 of flagellin was a dominant B-cell epitope at the surface of d flagella, because replacement of these two amino acids alone or together with their flanking sequence by a tripeptide specified by a linker sequence eliminated most reactivity with antisera against wild-type d flagella as tested by enzyme-linked immunosorbent assay or by Western immunoblot . Functional analysis of the mutated flagellin genes with or without an insert suggested that amino acids 180 to 214 in the 5' part of hypervariable region IV (residues 181 to 307 of the total of 505) is important to the function of flagella . The hybrid proteins formed by insertion of peptide sequence pre-S1 12-47 of hepatitis B virus surface antigen into the deleted flagellins assembled into functional flagella, and antibody to the pre-S1 sequence was detected after immunization of mice with the hybrid protein . This suggests that such mutant flagellins containing heterologous epitopes have potential as vaccines. Mutat Res, 1994 Apr, 317(2), 89 - 109 Experimental databases on inhibition of the bacterial mutagenicity of 4-nitroquinoline 1-oxide and cigarette smoke; Camoirano A et al.; Two antimutagenicity databases were prepared by applying a co-treatment procedure to the Salmonella reversion assay . Ninety compounds belonging to various chemical classes were quantitatively tested for antimutagenicity towards the direct-acting mutagen 4-nitroquinoline 1-oxide (4NQO) in strain TA100 of S . typhimurium and 63 of them were additionally tested for antimutagenicity towards unfractionated mainstream cigarette smoke (CS) in strain TA98, in the presence of S9 mix . Twelve compounds (13.3%) inhibited 4NQO mutagenicity by at least 50%, with a MID50 (dose inhibiting 50% of mutagenicity) varying over a 1226-fold range . Twenty-six compounds (41.3%) inhibited CS mutagenicity, with a MID50 varying over a 520-fold range . Three compounds only, i.e., bilirubin, curcumin and myricetin, were capable of inhibiting the mutagenicities of both 4NQO and CS . However, myricetin and the other flavonoid rutin were at the same time mutagenic by inducing frameshift mutations following metabolic activation . There was a rather rigorous selectivity of antimutagenicity data depending on the chemical class of inhibitors and it was possible to discriminate protective effects within several pairs or series of structurally related compounds . For instance, all eight thiols and aminothiols inhibited 4NQO mutagenicity, which contrasted with the inactivity of the remaining 17 sulfur compounds tested, all of them lacking a free sulfhydryl group . The mutagenicity of CS was consistently inhibited by the majority of phenols (eight out of 10 tested) and by all two isothiocyanates, two dithiocarbamates, three indole derivatives, three tetrapyrrole compounds and three flavonoids tested . Although the results obtained cannot be extrapolated to other mutagens or test systems, they may provide a useful source of information for research in the area of antimutagenesis and for the development of chemopreventive agents. Mutat Res, 1994 Apr, 317(2), 111 - 32 Ethylene thiourea (ETU) . A review of the genetic toxicity studies; Dearfield KL; Ethylene thiourea (ETU) is a common contaminant, metabolite and degradation product of the fungicide class of ethylene bisdithiocarbamates (EBDCs); as such, they present possible exposure and toxicological concerns to exposed individuals . ETU has been assayed in many different tests to assess genotoxicity activity . While a great number of negative results are found in the data base, there is evidence that demonstrates ETU is capable of inducing genotoxic endpoints . These include responses for gene mutations (e.g . Salmonella), structural chromosomal alterations (e.g . aberrations in cultured mammalian cells as well as a dominant lethal assay) and other genotoxic effects (e.g . bacterial rec assay and several yeast assays) . It is important to consider the magnitude of the positive responses as well as the concentrations/doses used when assessing the genotoxicity of ETU . While ETU induces a variety of genotoxic endpoints, it does not appear to be a potent genotoxic agent . For example, it is a weak bacterial mutagen in the Salmonella assay without activation in strain TA1535 at concentrations generally above 1000 micrograms/plate . Weak genotoxic activity of this sort is usually observed in most of the assays with positive results . Since ETU does not appear very potent and is not extremely toxic to test cells and organisms, it is not surprising to find that ETU does not produce consistent effects in many of the assays reviewed . Consequently, in many instances, mixed results for the same assay type are reported by different investigators, but as reviewed herein, these results may be dependent upon the test conditions in each individual laboratory . A primary shortcoming with many of the reported negative results is that the concentrations or doses used are not high enough for an adequate test for ETU activity . There are also problems with many of the negative assays generally in protocol or reporting, particularly with the in vivo studies (e.g . inappropriate sample number and/or sampling times; inadequate top dose employed) . Overall, while ETU does not appear to be a potent genotoxic agent, it is capable of producing genotoxic effects (e.g . gene mutations, structural chromosomal aberrations) . This provides a basis for weak genotoxic activity by ETU . Furthermore, based on a suggestive dominant lethal positive result, there may be a concern for heritable effects . Due to the many problems with the conduct and assessment of the in vivo assays, it is worth repeating in vivo cytogenetic assays and a dominant lethal assay (with acceptable test procedures and data generation) to determine if these results would continue to support a heritable mutagenicity concern. Mutat Res, 1994 Apr, 321(1-2), 43 - 56 Evaluation of carbendazim for gene mutations in the Salmonella/Ames plate-incorporation assay: the role of aminophenazine impurities; Sarrif AM et al.; Benomyl (methyl {1-{(butylamino)carbonyl}-1H-benzimidazol-2- yl}carbamate) and its major metabolite carbendazim (methyl 2-benzimidazolecarbamate) are major agricultural systemic fungicides . These compounds inhibit fungal microtubular function and thereby cause nondisjunction of chromosomes at cell division . Several investigators have proposed that these compounds can also cause gene mutations (base-pair substitutions) . In this laboratory, no mutagenic activity was observed with either benomyl (analytical grade) or Benlate (samples tested up to 500 and 1200 micrograms/plate, respectively, the limit of cytotoxicity) in the Salmonella/Ames plate-incorporation test in either base-pair substitution (TA100 and TA1535) or frameshift-sensitive (TA98 and TA1537) strains with or without S9 metabolic activation . However, some carbendazim preparations caused mutations in frameshift-sensitive strains at very high concentrations (> or = 5000 micrograms/plate) with metabolic activation . The mutagenic activity was not due to the major carbendazim metabolite, methyl (5-hydroxy-1H-benzimidazol-2-yl)carbamate (5-OH MBC), since 5-OH MBC was not mutagenic with (up to 20,000 micrograms/plate) or without (up to 16,000 micrograms/plate) activation . Subsequently, two highly mutagenic contaminants, 2,3-diaminophenazine (DAP) and 2-amino-3-hydroxyphenazine (AHP) were detected in mutagenic carbendazim samples . In those samples, DAP and AHP contaminant levels ranged as high as 46.5 and 11.6 ppm, respectively . No evidence of mutagenicity could be detected in preparations in which the DAP content was < 1.8 ppm . The mutagenic activity of these two contaminants was further investigated in strain TA98 . Without activation, DAP and AHP were positive at test concentrations as low as 5 and 10 micrograms/plate, respectively . In the presence of S9, mutations were detected at much lower concentrations (beginning at 0.025 and 0.05 microgram/plate, respectively) . These results indicate that carbendazim samples containing DAP or AHP at levels as low as 5 or 10 ppm, respectively, would be positive in the Salmonella/Ames test with activation when tested at 5000 micrograms/plate . Purified carbendazim is not mutagenic. Proc Natl Acad Sci U S A, 1994 Mar 29, 91(7), 2552 - 6 Recombinational basis of serovar diversity in Salmonella enterica; Li J et al.; The fliC gene, which encodes phase 1 flagellin, was sequenced in strains of 15 Sa