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mass production of sphingomyelinase of bacillus cereus by a protein-hyperproducing strain, bacillus brevis 47, and its purification.sphingomyelinase (sphingomyelin cholinephosphohydrolase) [ec 3.1.4.12] of bacillus cereus was overproduced in a protein-hyperproducing strain, b. brevis 47, by cloning the gene into an expression vector pnu211, which has been developed to express a foreign gene utilizing a promoter and a signal sequence of an outer cell wall protein gene. from 1 liter of culture, about 10 mg of protein was purified to near-homogeneity by two steps of column chromatography; this is almost 500 times higher product ...19921491003
relationship between selected bacteria and the growth of immature house flies, musca domestica, in an axenic test system.to investigate the relationship between immature (maggot) house flies, musca domestica, and bacteria, we compared the development of sterile first-instar maggots in each of 10 pure blood agar cultures of bacteria with growth on sterile blood agar (negative control) and on standard house fly rearing medium (positive control). nine species of bacteria representing gram-negative and gram-positive rods, coccoid, and micrococcoid cell types supported house fly growth on blood agar. one bacterium, a s ...19921495035
the role of the pro-sequence in the processing and secretion of the thermolysin-like neutral protease from bacillus cereus.the bacillus cereus cnp gene coding for the thermolysin-like neutral protease (tnp) has been cloned, sequenced, and expressed in bacillus subtilis. the protease is first produced as a pre-pro-protein (m(r) = 61,000); the pro-peptide is approximately two-thirds of the size of the mature protein. the pro-sequence has been compared with those of six other tnps, and significant homologies have been found. additionally, the tnp pro-sequences are shown to be homologous to the pro-sequence of pseudomon ...19921495388
[meningitis caused by bacillus cereus]. 19921498184
early diagnosis of infectious keratitis with in vivo real time confocal microscopy.the tandem scanning confocal microscope (tsm) was adapted for in vivo examination of the cornea in rabbits with experimental bacterial and fungal keratitis. compared to slit lamp biomicroscopy, the tsm provides superior lateral and axial resolution and serial optical sectioning capability, which may be useful for identification of corneal pathogens in the early stages of infection. we used the tsm to examine normal rabbit eyes infected with bacteria (bacillus cereus) and a filamentous fungus (as ...19921499129
[determining the sensitivity of anthrax bacteria to antibiotics for its differentiation from the antibiotic sensitivity of spore-forming saprophytes]. 19921514849
monoclonal antibody-beta-lactamase conjugates for the activation of a cephalosporin mustard prodrug.cephalosporin mustard (cm) was designed as an anticancer prodrug that could be activated in a site-specific manner by monoclonal antibody-beta-lactamase conjugates targeted to antigens present on tumor cell surfaces. purified beta-lactamases from bacillus cereus (bc beta l) and escherichia coli (ec beta l) catalyzed the release of phenylenediamine mustard (pdm) from cm through a fragmentation reaction which occurs after the beta-lactam ring of cm is hydrolyzed. the km and vmax values were 5.7 mi ...19921515470
evaluation of the effect of 3 probiotics on experimental escherichia coli enterotoxaemia in weaned piglets.in this report the efficacy of 3 different probiotics (bacillus cereus "toyoi", lactobacillus spp. and streptococcus faecium) was investigated. they were supplemented in the food of recently weaned piglets that were orally infected with e. coli o141 k85ab. supplementation could not prevent mortality and clinical symptoms nor reduce the faecal excretion of hemolytic e. coli. the possible explanations for the unsatisfying results are various.19921519408
purification of an enterotoxin produced by bacillus cereus by immunoaffinity chromatography using a monoclonal antibody.a murine monoclonal antibody (mab) with high reactivity to an enterotoxin produced by bacillus cereus was used to prepare an immunoadsorbent for purification of the enterotoxin. by immunoaffinity chromatography using the immunoadsorbent, approximately 25% of crude enterotoxin applied was recovered in the eluate. the purified enterotoxin was found to be electrophoretically and antigenically homogeneous. it also showed vascular permeability activity and mouse lethality, and caused fluid accumulati ...19921521189
effect of 22r-hydroxycholesterol on the action of sphingomyelinase from bacillus cereus toward bovine erythrocytes.the incorporation of 22r-hydroxycholesterol [(22r)-5-cholestene-3 beta,22-diol] into the bovine erythrocyte membranes remarkably enhanced the degradation of sphingomyelin in erythrocyte membranes by the action of sphingomyelinase from bacillus cereus, causing much faster hemolysis of erythrocytes. the stimulative effect of 22r-hydroxycholesterol on the breakdown of sphingomyelin was maximal in the presence of mg2+. on the other hand, in spite of the presence of 22r-hydroxycholesterol, the breakd ...19921523674
structure and mechanism of alkaline phosphatase.alkaline phosphatase was the first zinc enzyme to be discovered in which three closely spaced metal ions (two zn ions and one mg ion) are present at the active center. zn ions at all three sites also produce a maximally active enzyme. these metal ions have center-to-center distances of 3.9 a (zn1-zn2), 4.9 a (zn2-mg3), and 7.1 a (zn1-mg3). despite the close packing of these metal centers, only one bridging ligand, the carboxyl of asp51, bridges zn2 and mg3. a crystal structure at 2.0-a resolutio ...19921525473
nucleotide sequence and deduced functions of a set of cotranscribed genes of streptomyces coelicolor a3(2) including the polyketide synthase for the antibiotic actinorhodin.a 5.3-kb region of the streptomyces coelicolor actinorhodin gene cluster, including the genes for polyketide biosynthesis, was sequenced. six identified open reading frames (orf1-6) were related to genetically characterized mutations of classes acti, vii, iv, and vb by complementation analysis. orf1-6 run divergently from the adjacent actiii gene, which encodes the polyketide synthase (pks) ketoreductase, and appear to form an operon. the deduced gene products of orf1-3 are similar to fatty acid ...19921527048
interaction of energized bacteria cells with particles of colloidal gold: peculiarities and kinetic model of the process.it is found that the cells of bacillus cereus b-4368 at energized state can concentrate the colloidal gold particles on their surface. it is shown that the process depends on metabolic reactions proceeding on the plasma membrane. the inhibitory analysis permits to suppose that the metal concentration is due to the functioning of atp-dependent generator of the transmembrane potential, apparently, of proton atpase. kinetic characteristics of the process show the presence of an intermediate state i ...19921532513
assignment of bacillus thermoamyloliquefaciens kp1071 alpha-glucosidase i to an exo-alpha-1,4-glucosidase, and its striking similarity to bacillary oligo-1,6-glucosidases in n-terminal sequence and in structural parameters calculated from the amino acid composition.alpha-glucosidase i of bacillus thermoamyloliquefaciens kp1071 (ferm p8477, facultative thermophile) was purified to homogeneity. the relative molecular mass was estimated to be 62,000 da. from its catalytic properties, the enzyme has been assigned to an exo-alpha-1,4-glucosidase. the enzyme shares its antigenic groups in part with bacillus stearothermophilus atcc12016 (obligate thermophile) exo-alpha-1,4-glucosidase. these exo-alpha-1,4-glucosidases strikingly resemble oligo-1,6-glucosidases fr ...19921555585
adp-ribosylation of small gtp-binding proteins by bacillus cereus.a human pathogenic strain of bacillus cereus produces an exoenzyme which selectively adp-ribosylates 20-25 kda gtp-binding proteins in platelet membranes. pre-adp-ribosylation of rho proteins of human platelet membranes with clostridium botulinum exoenzyme c3 or clostridium limosum exoenzyme inhibits subsequent adp-ribosylation by the exoenzyme from b. cereus indicating similar substrate specificity of the transferases. the adp-ribosyltransferase from b. cereus reveals no immunological cross-rea ...19921567406
antimicrobial properties of secretions from the metapleural glands of myrmecia gulosa (the australian bull ant).myrmecia gulosa (australian bull ant) produce secretions from their metapleural exocrine glands which have broad spectrum antimicrobial properties. such secretions are probably of importance in disease control in bull ant communities. these antimicrobial secretions are stable at 100 degrees c, resistant to proteolytic enzymes and are active over a wide ph range. of the organisms tested only endospores of bacillus cereus were found to be resistant. the antimicrobial agent(s) are absorbed by cells ...19921568945
synthetic phospholipids as substrates for phospholipase c from bacillus cereus.the substrate requirement of phospholipids for hydrolysis with phospholipase c from bacillus cereus was studied with synthetic lipids well-defined in structure and configuration. for optimal activity, the glycerol molecule must contain three substituents: phosphocholine in sn-3-, an ester bond in sn-2- and an ether- or ester bond in sn-1-position. the length of the ester or ether chains is of minor importance. any deviation from these structural requirements results in a large decrease in the hy ...19921571359
formycins a and b and some analogues: selective inhibitors of bacterial (escherichia coli) purine nucleoside phosphorylase.formycin b (fb), a moderate inhibitor (ki approximately 100 microm) of mammalian purine nucleoside phosphorylase (pnp), and formycin a (fa), which is totally inactive vs. the mammalian enzyme, are both effective inhibitors of the bacterial (escherichia coli) enzyme (ki approximately 5 microm). examination of a series of n-methyl analogues of fa and fb led to the finding that n(6)-methyl-fa, virtually inactive vs. the mammalian enzyme, is the most potent inhibitor of e. coli purine nucleoside pho ...19921576149
bacillus cereus endocarditis: report of a case and review.bacillus cereus is a ubiquitous organism that infrequently causes serious infections. we report a patient with b. cereus endocarditis involving a mechanical aortic valve. data for 10 cases of b. cereus endocarditis reported in the literature are summarized. b. cereus is resistant to many commonly used antibiotics, a finding that has clinical significance for empirical antibiotic selection in patients with suspected endocarditis. infection in patients with valvular heart disease in the few cases ...19921576291
toxic properties of bacillus cereus strains isolated from different foodstuffs.toxic activity of 89 culture filtrates of bacillus cereus strains were examined. from the total number of the filtrates tested, 3 were found to cause fluid accumulation in the ligated rabbit ileal loop. 71 samples caused a vascular permeability increase in the rabbit skin corresponding to an intermediate or strong toxin production. 59 of the 89 culture filtrates were cytotoxic on vero cells, 31 had a lethal effect, 81 induced hemolysis of 0.5% rabbit erythrocytes and 55 tested samples showed lec ...19921576401
substrate-enzyme interactions and catalytic mechanism in phospholipase c: a molecular modeling study using the grid program.based on the high-resolution x-ray crystallographic structure of phospholipase c from bacillus cereus, the orientation of the phosphatidylcholine substrate in the active site of the enzyme is proposed. the proposal is based on extensive calculations using the grid program and molecular mechanics geometry relaxations. the substrate model has been constructed by successively placing phosphate, choline and diacylglycerol moieties in the positions indicated from grid calculations. on the basis of th ...19921579567
role of spore coats in the germinative response of bacillus cereus to adenosine and its analogues.spores of the strain ncib 8122 of bacillus cereus have been depleted of coats by treatment with 0.1% sodium dodecyl sulfate--200 mm 2-mercaptoethanol--0.5 m nacl (ph 9.6). the coat-depleted spores did not show any decrease in viability, heat resistance, refractility, dipicolinic acid content, or specific activities of several protoplastic enzymes. the germinative response of the coat-depleted spores to adenosine and several analogues thereof was found qualitatively similar to that obtained with ...19921581864
a type-iii dna restriction and modification system in bacillus cereus? 19921587478
physical maps of the genomes of three bacillus cereus strains.noti restriction maps of the chromosomes from bacillus cereus atcc 10876, atcc 11778, and the b. cereus type strain atcc 14579 have been established and compared with the previously established map of b. cereus atcc 10987. between 10 and 14 noti fragments were observed, ranging from 15 to 1,300 kb, in digests of dna from the various strains. the sizes of the genomes varied between 5.4 and 6.3 mb. the maps were constructed by hybridization of 42 random probes, prepared from b. cereus atcc 10987 l ...19921592826
crystal structures of phosphate, iodide and iodate-inhibited phospholipase c from bacillus cereus and structural investigations of the binding of reaction products and a substrate analogue.the crystal structure of the complex formed between phospholipase c (plc) from bacillus cereus and inorganic phosphate (pi), which is an inhibitor, has been determined and refined to 2.1 a resolution. the final r-factor is 19.7%. we have also studied the binding of two other inhibitors, iodide and iodate, to plc. x-ray data for these two complexes were collected to 2.8 a resolution during the search for heavy-atom derivatives. a series of screening experiments where plc crystals have been treate ...19921593635
investigation of the substrate binding and catalytic groups of the p-c bond cleaving enzyme, phosphonoacetaldehyde hydrolase.kinetic studies with substrate analogs and group-directed chemical modification agents were carried out for the purpose of identifying the enzyme-substrate interactions required for phosphonoacetaldehyde (p-ald) binding and catalyzed hydrolysis by p-ald hydrolase (phosphonatase). malonic semialdehyde (ki = 1.6 mm), phosphonoacetate (ki = 10 mm), phosphonoethanol (ki = 10 mm), and fluorophosphate (ki = 20 mm) were found to be competitive inhibitors of the enzyme but not substrates. thiophosphonoa ...19921605625
bacteriological quality of infant feeding bottle-contents and teats in addis abeba, ethiopia.bacteriological quality of 244 infant feeding-bottles and 61 teats were studied from january 1989 to november 1989. samples were collected from feeding-bottles of babies who were brought by nursing mothers to clinics and hospitals for varying complaints. of the 244 bottle-content samples, 144 (59%) harboured more than 2 x 10(6) colonies of bacteria per ml. of samples. the predominant bacterial isolates both from bottle-contents and teat-swabs were the coliform group of bacteria, such as e. coli, ...19921606947
isolation and characterisation of restriction endonuclease bcuai from bacillus cereus a. 19921614885
rapid identification of viable bacterial spores using a fluorescence method.a method has been devised to differentiate viable and nonviable bacterial spores. "germination-like" changes are initiated in spores with performic acid and lysozyme. the germinated spores are stained with aqueous acridine orange, a fluorescent dye. nonviable spores fluoresce lemon-green and viable spores orange-red. it is proposed that with the use of a membrane filter resistant to performic acid and lysozyme, the method may be used for spore enumeration in foods in about 4 hr compared to conve ...19921616999
nucleotide sequence and phylogeny of the tet(l) tetracycline resistance determinant encoded by plasmid pste1 from staphylococcus hyicus.the nucleotide sequence of the tetracycline resistance (tet) gene and its regulatory region, encoded by the plasmid pste1 from staphylococcus hyicus, was determined. the tet gene was inducible by tetracycline and encoded a hydrophobic protein of 458 amino acids. comparisons between the predicted amino acid sequences of the pste1-encoded tet from s. hyicus and the previously sequenced tet k variants from staphylococcus aureus, tet l variants from bacillus cereus, bacillus stearothermophilus, and ...19921622166
campylobacter jejuni non-culturable coccoid cells.the behaviour of campylobacter jejuni in the environment is poorly documented. rapid loss of viability on culture media is reported. this phenomenon is associated with the development of so-called coccoid cells. it has been suggested that these cells can be infective to animals and man. results obtained with atp-measurements of coccoid cells and direct viable count (dvc) support this hypothesis. introduction of coccoid cells into simulated gastric, ileal and colon environments did not result in ...19921622752
tracking laboratory contamination by using a bacillus cereus pseudoepidemic as an example.from 1 march to 31 may 1990, bacillus cereus was recovered from 24 of 5,534 (0.49%) blood cultures and 22 of 1,088 (2.02%) other body fluid cultures. the rarity of this organism as a pathogen and comparison with previous baseline rates led to the conclusion that it was a pseudoepidemic involving some form of culture contamination. generalized precautions taken without specific knowledge of the contaminant source reduced the recovery rate of the organism. recovery rates for the organism returned ...19921624565
transfer of a chloroplast-bound precursor protein into the translocation apparatus is impaired after phospholipase c treatment.we have studied the influence of phospholipase c treatment of intact purified chloroplast on the translocation of a plastid destined precursor protein. under standard import conditions, i.e. in the light in the presence of 2 mm atp translocation was completely abolished but binding was observed at slightly elevated levels. an experimental regime which allowed binding but not import of the precursor protein, i.e. in the dark in the presence of 10 microm atp, demonstrated that translocation interm ...19921628746
quantification of distinct molecular species of the 2-lyso metabolite of platelet-activating factor by gas chromatography-negative-ion chemical ionization mass spectrometry.the biological activity of platelet-activating factor (paf) is comprised by a few molecular species of phosphatidylcholine which contain a fatty alcohol connected by an ether linkage to the sn-1 position of the glycerol backbone and an acetate ester at the sn-2 position. the various molecular species of paf differ in chain length and degree of unsaturation in the fatty alcohol residue side-chain. paf is rapidly hydrolyzed to lyso-paf by an acetylhydrolase enzyme which is quite active in a number ...19921629294
differential scanning calorimetric study of the thermal unfolding of beta-lactamase i from bacillus cereus.the irreversible thermal unfolding of the class a beta-lactamase i from bacillus cereus has been investigated at ph 7.0, using differential scanning calorimetry (dsc) and inactivation kinetic techniques. dsc transitions showed a single peak with a denaturation enthalpy of 646 kj.mol-1 and were moderately scan rate dependent, suggesting that the process was partially kinetically controlled. the inactivation kinetics at constant temperature showed that the irreversible denaturation of the enzyme o ...19921633173
the structure of neutral protease from bacillus cereus at 0.2-nm resolution.the crystal structure of the neutral protease from bacillus cereus has been refined to an r factor of 17.5% at 0.2-nm resolution. the enzyme, an extracellular metalloendopeptidase, consists of two domains and binds one zinc and four calcium ions. the structure is very similar to that of thermolysin, with which the enzyme shares 73% amino-acid sequence identity. the active-site cleft between the two domains is wider in neutral protease than in thermolysin. this suggests the presence of a flexible ...19921633827
a single mode fibre-optic evanescent wave biosensor.this paper reports experimental developments in the construction and operation of a single-mode fibre-optic evanescent wave biosensor using an exposed core silica single-mode fibre embedded in a silica block. the device was able to monitor the concentration of a blue dye, procion blue mx-g, in overlayers of various refractive indices. the practicality of such a biosensor has been demonstrated with a colorimetric enzyme assay system. penicillin g in the 0-0.4 mm concentration range was monitored ...19921637526
an outbreak of bacillus cereus food poisoning--are caterers supervised sufficiently.bacillus cereus is an uncommonly reported cause of foodborne illness in the united states. in may 1989, an outbreak of b. cereus gastroenteritis occurred among 140 guests who had attended a catered wedding reception in napa, ca. investigation established cornish game hens served at the event as the vehicle for disease transmission (or = 29, p = 0.0001). although the spores of b. cereus are ubiquitous, large numbers of toxin-producing organisms (more than 10(5) per gram of food) are required for ...19921641447
production of the vacuolation factor of bacillus cereus isolated from vomiting-type food poisoning.vacuole response in hep-2 cells was induced with culture supernatants of bacillus cereus strains isolated from outbreaks of vomiting- and diarrheal-type food poisoning grown in rice flour and laboratory media. high vacuole response was obtained with culture supernatants of b. cereus strains isolated from vomiting-type food poisoning grown in cooked rice suspension or on a cooked rice plate, whereas no response was obtained with those of the same strains grown in brain heart infusion and trypto-s ...19921643168
an investigation of dye reduction by food-borne bacteria.the rates of reduction of seven redox dyes by 13 bacterial strains were measured and found to vary greatly between different bacterium/dye combinations. phenazine ethosulphate and toluidine blue were the most rapidly reduced dyes by the majority of bacteria and resorufin and 2-hydroxy-1,4-naphthoquinone were reduced slowly, if at all. there was also considerable variation in the rates of reduction with any single dye/organism combination. glucose stimulated the rates of endogenous dye reduction ...19921644703
identification of phosphatidylinositol-specific phospholipase c activity in listeria monocytogenes: a novel type of virulence factor?a phospholipase c which cleaves phosphatidylinositol and glycosylphosphatidylinositol (gpi) anchors was identified in listeria monocytogenes. this 36 kda protein is encoded by the gene plca, and is homologous to the bacillus cereus, bacillus thuringiensis and eukaryotic phosphatidylinositol-specific phospholipases c (pi-plc). expression of the plca gene in escherichia coli correlates with the appearance of pi-plc activity in the cells. in listeria monocytogenes, the activity is secreted to the c ...19911645839
phosphatidylinositol-specific phospholipases c from bacillus cereus and bacillus thuringiensis. 19911646937
beta-lactamase inhibiting properties of 6-oxopenicillanic and 7-oxo-cephalosporanic acid derivatives. 19911649611
the possible involvement of trypsin-like enzymes in germination of spores of bacillus cereus t and bacillus subtilis 168.germination of spores of bacillus cereus t and bacillus subtilis 168 was inhibited by the trypsin inhibitors leupeptin and tosyllysine chloromethyl ketone (tlck) and by the substrates tosylarginine methyl ester (tame), benzoyl-l-arginine-p-nitroanilide (l-bapna) and d-bapna. potencies of these inhibitory compounds were estimated by finding the concentration which inhibited 50% germination (id50), as measured by events occurring early (loss of heat resistance), at an intermediate stage [dipicolin ...19911650815
two different aa3-type cytochromes can be purified from the bacterium bacillus cereus.two aa3-type cytochromes were purified from membranes of sporulating bacillus cereus. one of them, an aa3 complex, was found to be composed of two subunits (51 and 31 kda), two a hemes and three copper atoms, thus being similar to the cytochrome aa3 previously purified from vegetative b. cereus [garcía-horsman, j. a., barquera, b., gonzález-halphen, d. & escamilla, j. e. (1991) mol. microbiol. 5, 197-205]. the second isoform, a caa3 complex, was expressed in sporulating cells only, and was found ...19911651246
role of gtpase activating protein in mitogenic signalling through phosphatidylcholine-hydrolysing phospholipase c.recent evidence has accumulated showing that activation of plc-catalysed hydrolysis of phosphatidylcholine (pc-plc) is a critical step in mitogenic signal transduction both in fibroblasts and in oocytes from xenopus laevis. the products of ras genes activate pc-plc, bind guanine nucleotides, have intrinsic gtpase activity, and are regulated by a gtpase-activating protein (gap). it has been suggested that, in addition to its regulatory properties, gap may also be necessary for ras function as a d ...19911655413
a surface antigen of giardia lamblia with a glycosylphosphatidylinositol anchor.since giardia lamblia trophozoites are exposed to high concentrations of fatty acids in their human small intestinal milieu, we determined the pattern of incorporation of [3h]palmitic acid and myristic acid into g. lamblia proteins. the pattern of fatty acylation was unusually simple since greater than 90% of the giardia protein biosynthetically labeled with either [3h]palmitate or myristate migrated at approximately 49 kda (gp49) in reducing sodium dodecyl sulfate-polyacrylamide gel electrophor ...19911657957
glycosyl-phosphatidylinositol anchored acetylcholinesterase as substrate for phosphatidylinositol-specific phospholipase c from bacillus cereus.we investigated the enzymatic properties of phosphatidylinositol-specific phospholipase c (pi-plc) from bacillus cereus towards glycosyl-phosphatidylinositol anchored acetylcholinesterase (ache) from bovine erythrocytes and torpedo electric organ as substrate. the conversion of membrane from ache to soluble ache by pi-plc depended on the presence of a detergent and of phosphatidylcholine. in presence of mixed micelles containing triton x-100 (0.05%) and phosphatidylcholine (0.5 mg/ml) the rate o ...19911660725
high-level expression in escherichia coli and rapid purification of phosphatidylinositol-specific phospholipase c from bacillus cereus and bacillus thuringiensis.the construction of four vectors for high-level expression in escherichia coli of the phosphatidylinositol-specific phospholipase c from bacillus cereus or bacillus thuringiensis is described. in all constructs the coding sequence for the mature phospholipase is precisely fused to the e. coli heat-stable enterotoxin ii signal sequence for targeting of the protein to the periplasm. in one set of plasmids expression of the b. cereus or b. thuringiensis enzyme is under control of the e. coli alkali ...19911668269
fried rice from the take-away. 19911669788
purification and characterization of a repressor for the bacillus cereus glnra operon.we report the overexpression, purification, and properties of the regulatory protein, glnr, for glutamine synthetase synthesis of bacillus cereus. the protein was found to be a dimer with a molecular weight of approximately 30,000, and its subunit molecular weight was 15,000 in agreement with that (15,025) of deduced amino acid sequence of glnr. the purified glnr protein bound specifically to the promoter region of the glnra operon of b. cereus and bacillus subtilis. the binding of the glnr prot ...19911677938
detection of bacillus cereus flagellar antigen by enzyme-linked immunosorbent assay (elisa)a serological typing scheme of bacillus cereus has been developed by immunochemical analyses of flagellar antigen using an agglutination method. enzyme-linked immunosorbent assay (elisa) for the classification of flagellar serotype of bacillus cereus had greater sensitivity. 10-500 times, than that of agglutination method. the specificity of flagellar antigen and antibody was determined by immunogold electron microscopy and elisa inhibition assay. application of elisa is useful for the detection ...19911678490
inhibition of the phosphatidylinositol-specific phospholipase c from bacillus cereus by a monoclonal antibody binding to a region with sequence similarity to eukaryotic phospholipases.bacterial phosphatidylinositol-specific phospholipases c (pi-plc) display similar substrate specificity as their eukaryotic counterparts involved in signal transduction of insulin and ca2(+)-mobilizing hormones, and are used in the study of the novel glycosylphosphatidylinositol-protein anchors (gpi-anchors). for the investigation of structure-function aspects of the pi-plc secreted from bacillus cereus cells, a panel of murine monoclonal antibodies was generated and shown to be specific for the ...19901701099
purification and characterization of neuron-specific surface antigen defined by monoclonal antibody bm88.monoclonal antibody bm88 recognizes a neurospecific surface antigen in the cns and the pns. in the present study, the antigen recognized by bm88 was immunopurified from pig brain and shown to be a 22-kda polypeptide by reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. under nonreducing conditions a protein of 40 kda was obtained, a result indicating that the antigen is composed of two polypeptide chains of equal molecular weight linked by disulfide bridges. gel filtration of th ...19911704420
crystal structure of penicillium citrinum p1 nuclease at 2.8 a resolution.p1 nuclease from penicillium citrinum is a zinc dependent glyco-enzyme consisting of 270 amino acid residues which cleaves single-stranded rna and dna into 5'-mononucleotides. the x-ray structure of a tetragonal crystal form of the enzyme with two molecules per asymmetric unit has been solved at 3.3 and refined at 2.8 a resolution to a crystallographic r-factor of 21.6%. the current model consists of 269 amino acid residues, three zn ions and two n-acetyl glucosamines per subunit. the enzyme is ...19911710977
comparative analysis of bacillus anthracis, bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16s rrna.the primary structures of the 16s rrnas of bacillus anthracis, bacillus cereus, bacillus mycoides, and bacillus thuringiensis were determined by using the reverse transcription-dideoxy sequencing method. all of the strains exhibited very high levels of sequence similarity (greater than 99%) that were consistent with the close relationships shown by previous dna hybridization studies. the sequences of b. anthracis sterne and b. cereus emetic strain nctc 11143 were found to be identical for a cont ...19911715736
lipid globule staining to aid in differentiating bacillus species.the use of the lipid globule stain to aid in differentiating the bacillus cereus group (i.e., b. cereus, b. cereus var. mycoides, and b. thuringiensis) from other bacillus species was investigated. smears from colonies grown on suitable agar were made on precleaned slides, stained, and examined microscopically for characteristic deep blue lipid globules. the study included a total of 649 cultures of bacillus species plus 143 incompletely characterized bacillus isolates from food. only b. cereus, ...19911717433
blasticidin s-resistance gene (bsr): a novel selectable marker for mammalian cells.blasticidin s is a microbial antibiotic that inhibits protein synthesis in both prokaryotes and eukaryotes. the blasticidin s-resistance gene (bsr), isolated from bacillus cereus k55-s1 strain, was inserted into psv2 plasmid vector and introduced into cultured mammalian cells by transfection. the bsr gene was integrated into the genome and conferred blasticidin s resistance on hela cells. the transfection frequency of the bsr gene was as high as that of the aminoglycoside phosphotransferase gene ...19911720391
deoxyribose 5-phosphate aldolase of bacillus cereus: purification and properties.deoxyribose 5-phosphate aldolase was purified 41 times from bacillus cereus induced by growth on deoxyribonucleosides. the purification procedure includes ammonium sulphate fractionation, gel filtration on sephadex g-100, ion-exchange chromatography on deae-sephacel and preparative electrophoresis on 10% polyacrylamide gel. the enzyme is stable above ph 6.5, but is rapidly inactivated by sulfhydryl reagents. being insensitive to edta, it may be considered as a class i aldolase. among a number of ...19921730028
critical micelle concentration and hemolytic activity--a correlation suggested by the marine sterol, halistanol trisulfate.the marine natural product, halistanol trisulfate, has a relatively low critical micelle concentration of 0.001% m/v (14.5 microm) and strong hemolytic potency with an ec50 of 0.00046% m/v (6.67 microm). as expected of a detergent, it inhibits the growth of gram-positive but not gram-negative bacteria. the hemolytic activity of halistanol trisulfate and other detergents has been shown to correlate with critical micelle concentration. this correlation may have important implications in the mechan ...19921731774
the effect of microwave and conventional cooking on the temperature profiles and microbial flora of minced beef.beef mince samples were cooked to the rare, medium and well done states by microwave and conventional oven methods. the raw samples all contained large numbers of contaminating organisms and low numbers of listeria monocytogenes were detected in the majority of samples. a substantial proportion of the contaminating flora survived in mince cooked by microwaves to the rare, medium and well done states, prior to a 30-min standing period. listeria monocytogenes survived in all samples cooked by micr ...19911742174
microbiological analysis and starter culture growth in retentates.pasteurized skim milk was concentrated by uf to 2-, 4-, and 5-fold. the retentates were evaluated for microbiological quality, heat treatments to inactivate microorganisms, and lactic acid bacterial starter culture activity. aerobic mesophilic bacterial counts in raw milk decreased from an initial 1.4 x 10(6) to 3.9 x 10(2) cfu/ml after pasteurization. during uf, counts increased from 3.9 x 10(2) cfu/ml uf, counts increased from 3.9 x 10(2) cfu/ml in pasteurized milk to 1.4 x 10(3), 1.4 x 10(4), ...19911744259
amperometric monitoring of bacteria-induced milk acidity using a platinum disc microelectrode.the acidity induced by the action of bacteria in milk samples was monitored amperometrically by using a platinum microelectrode. the measurements were performed directly on commercial packs of milk, stored at 32 degrees c, and were continued for 9-10 d after inoculation. the data were compared with those obtained by measuring the ph of the samples and the results are discussed on the basis of the metabolism of each bacterial species. the effects of the following bacteria were examined: staphyloc ...19911759720
proline residues responsible for thermostability occur with high frequency in the loop regions of an extremely thermostable oligo-1,6-glucosidase from bacillus thermoglucosidasius kp1006.the gene encoding for an extremely thermostable oligo-1,6-glucosidase from bacillus thermoglucosidasius kp1006 (dsm2542, obligate thermophile) was sequenced. the amino acid sequence deduced from the nucleotide sequence of the gene (1686 base pairs) corresponded to a protein of 562 amino acid residues with a mr of 66,502. its predicted amino acid composition, mr, and n-terminal sequence of 12 residues were consistent with those determined for b. thermoglucosidasius oligo-1,6-glucosidase. the dedu ...19911761534
enhancement of soybean nodulation by bacillus cereus uw85 in the field and in a growth chamber.seed treatments with bacillus cereus uw85 increased nodulation of soybeans in three field seasons and in three different sterilized soils in the growth chamber. in the field, 28 and 35 days after planting, uw85-treated plants had 31 to 133% more nodules than untreated plants. from 49 days after planting until seed harvest, there were no significant differences between nodulation of uw85-treated plants and untreated control plants. in the growth chamber, in sterilized soil-vermiculite mixtures, a ...19911768151
[effects of sodium chloride on destruction of microorganisms by microwave heating in potatoes].to assess the destructive effect of different sodium chloride concentrations (0, 0.3, 1.5, and 3%) on microorganisms with microwave heating, strains of vibrio parahaemolyticus, staphylococcus aureus, salmonella enteritidis, escherichia coli and bacillus cereus were used to inoculate a mashed potato preparation. after microwave heating for 1 min at 800 w, resulted in temperatures and rate of destruction of bacteria significantly higher (p less than 0.05) in the core than on the surface of mashed ...19911773068
purification and some properties of phospholipase c from achromobacter xylosoxidans.a non-haemolytic phospholipase c (ec 3.1.4.3) was purified from the culture medium of achromobacter xylosoxidans with a 5% yield and a purification factor of 330. a combination of ultrafiltration, acetone precipitation and two subsequent affinity chromatographic steps was used. the affinity chromatography is a new application of 2-(4-aminophenylsulphonyl)ethyl-cellulose, a sorbent that has previously been used for the purification of phospholipase c from bacillus cereus. the purified enzyme gave ...19911783637
[effect of escherichia coli on growth of bacillus cereus].the effect of 101 escherichia coli strains on growth of 90 bacillus cereus strains on solid media was investigated. only 9 e. coli strains (in particular the colicin +-generating ones) were antagonistic towards b. cereus, giving distinct growth-inhibition zones around the colonies.19911788515
phospholipase c-sensitive factor vii complexes in dog plasma.we report the presence of a phospholipase c-sensitive activated factor vii complex in canine plasma after feeding a special diet. low levels of complex was observed in the fasting state. the response to feeding in terms of activated factor vii varied markedly among the dogs investigated.19911788825
a fluorescent substrate for the continuous assay of phosphatidylinositol-specific phospholipase c: synthesis and application of 2-naphthyl myo-inositol-1-phosphate.a fluorescent water-soluble substrate for phosphatidylinositol-specific phospholipase c was synthesized. the diacylglycerol moiety of the natural substrate, phosphatidylinositol, was replaced by the fluorescent moiety, 2-naphthol, resulting in the synthetic substrate, racemic 2-naphthyl myo-inositol-1-phosphate. the synthetic substrate provided a continuous fluorometric assay for the phosphatidylinositol-specific phospholipase c from bacillus cereus. initial rates of the cleavage of the 2-naphth ...19911789410
ph gradients through colonies of bacillus cereus and the surrounding agar.ph-sensitive microelectrodes, constructed with a tip diameter of about 4 microns, were deployed through 24 h and 48 h colonies of bacillus cereus incubated on cys medium (casamino acids, yeast extract, salts), with and without glucose. measurements of ph were used to construct ph profiles through the colony and the surrounding agar. ph gradients could be detected for at least 800 microns into the agar beneath a 24 h colony, and to approximately 10 mm horizontally away from the edge of the colony ...19911791442
results in the management of bacillus endophthalmitis.eighteen cases of culture-proven bacillus endophthalmitis were reviewed. the majority (13/18) were secondary to penetrating ocular trauma. others were secondary to corneal ulcer (1), postoperative (1), and endogenous (3). bacillus cereus was the most commonly identified species (12/18). antibiotic sensitivities demonstrated uniform sensitivity to vancomycin and the aminoglycosides. visual prognosis was generally poor; only two patients maintained 20/200 or better in the affected eye. fourteen ey ...19911792034
hydroxylation and glucoside conjugation in the microbial metabolism of the diterpene sclareol.1. the microbial metabolism of sclareol (1), a labdane diterpene ditertiary alcohol, was studied. preliminary screening identified a number of microorganisms capable of metabolizing sclareol. 2. preparative scale fermentation of growing cultures of bacillus cereus ui-1477 resulted in the production of seven metabolites which have been characterized as 3 beta-hydroxysclareol (2), 2 alpha-hydroxysclareol (3), 18-hydroxysclareol (4), 2 alpha,18-dihydroxysclareol (6), 8 alpha,13 beta-dihydroxy-labd- ...19911796608
isolation and purification of bacterial proteinases by means of autofocusing.autofocusing was used for the isolation and purification of neutral and alkaline proteinases from fermentation broth, after separation of cells. the yield of proteinases achieved was 19-78%, and was inversely proportional to the degree of purification, which varied from 3.0 to 7.9. because of considerable losses of enzyme activity and the long duration of the process, autofocusing seems to be a non-economic technique for industrial isolation of relatively cheap enzymes.19911804847
a chromogenic substrate for phosphatidylinositol-specific phospholipase c: 4-nitrophenyl myo-inositol-1-phosphate.a chromogenic water-soluble substrate for phosphatidylinositol-specific phospholipase c was synthesized starting from myo-inositol employing isopropylidene and 4-methoxytetrahydropyranyl protecting groups. in this analogue of phosphatidylinositol, 4-nitrophenol replaces the diacylglycerol moiety, resulting in synthetic, racemic 4-nitrophenyl myo-inositol-1-phosphate. using this synthetic substrate a rapid, convenient and sensitive spectrophotometric assay for the phosphatidylinositol-specific ph ...19911814635
role of side chain moiety in the hydrolysis of penicillins by beta-lactamase.various beta-lactam compounds and structurally related moieties were examined as substrates of beta-lactamase from bacillus cereus 5/b nctc 9946. the enzyme was specific for penicillins and none of the cephalosporins were hydrolysed. electronic environment of allylic carboxy group in dihydrothiazine ring restricts the acceptance of cephalosporins as substrates. the efficiency of hydrolysis of penicillins is dependent on dense resonating electronic environment of phenyl ring present in the side c ...19911814862
hyperexpression in escherichia coli, purification, and characterization of the metallo-beta-lactamase of bacillus cereus 5/b/6.we used site-directed mutagenesis to introduce both a ndei restriction endonuclease site and an initiator codon at the junction of the leader and structural gene sequences of the metallo-beta-lactamase of bacillus cereus 5/b/6. this construct allowed us to clone just the beta-lactamase structural gene sequence into an escherichia coli expression vector. e. coli cells were transformed with the recombinant plasmid, the b. cereus beta-lactamase was expressed, and these e. coli cells were disrupted ...19911821784
fluid accumulation in mouse ligated intestine inoculated with the vascular permeability factor produced by bacillus cereus.partially purified vascular permeability (vp) factor (vpf) of bacillus cereus induced fluid accumulation in the ligated intestinal loops of mouse (mil) and rabbit (ril), suggesting that the vp activity may correlate with fluid accumulation in ligated intestinal loops of these animals. fluid accumulation was observed at 6-8 hr in 55-67% of mouse intestinal loops inoculated with 40-50 immunodiffusion units (idu) of partially purified vpf, whereas it was found at 2 hr in all loops with 400-600 idu ...19911830786
purification and characterization of the vascular permeability factor produced by bacillus cereus.purification of an extracellular protein exhibiting the vascular permeability activity produced by bacillus cereus was performed by ammonium sulfate precipitation followed by chromatography on de-32 cellulose, sephadex g-100, and sephadex g-75. the purified protein was found to be electrophoretically and antigenically almost homogeneous although it contained a trace of contaminant. the molecular weight of the protein was calculated to be 45,000 by polyacrylamide gel electrophoresis in the presen ...19911830799
in vitro stability in biological activity and antigenicity of the vascular permeability factor produced by bacillus cereus. 19911830805
development of murine monoclonal antibodies against an enterotoxin produced by bacillus cereus.three murine monoclonal antibodies (mabs) were prepared against an enterotoxin (et) produced by bacillus cereus. although these mabs were found to react with the et, their specificities appeared to be different in competitive binding assays. one of the mabs (d-8), which was highly reactive with the et, will be useful in developing immunological methods to detect crude et and to isolate the et in high yield.19911832030
purification and some properties of a bacillus cereus mouse lethal toxin.a mouse lethal toxin (mlt) produced by bacillus cereus isolated in vomiting-type food poisoning was purified by chromatography on deae-sephadex a-25 followed by gel filtration on sephadex g-75. purified mlt possessed a molecular weight of 33,000-34,000. it showed mouse lethality and hemolytic (hl) activity on sheep and rabbit erythrocytes; the latter erythrocytes were more weakly hemolyzed than the former ones. however, fluid accumulation in mouse ligated intestinal loops was not induced by puri ...19911832034
adsorption on bacterial spores depending on the aggregation properties of antimicrobial tensides.a change in interaction with spores of bacillus cereus occurred in the range of critical concentrations of micelle formation. with 1-methyldodecyldimethylamine-n-oxide and n,n'-bis(dodecyldimethyl)-1,2-ethanediammonium dibromide, the induced release of dipicolinic acid was blocked and the adsorption dynamics changed, respectively.19911841878
mechanism of inhibition of adenylate cyclase by phospholipase c-catalyzed hydrolysis of phosphatidylcholine. involvement of a pertussis toxin-sensitive g protein and protein kinase c.the phospholipase c-mediated hydrolysis of phosphatidylcholine has been shown recently to be activated by a number of agonists. muscarinic receptors, which trigger various signal transduction mechanisms including inhibition of adenylate cyclase through gi, have been shown to be potent stimulants of this novel phospholipid degradative pathway. we demonstrate here, by exogenous addition of bacillus cereus phosphatidylcholine-hydrolyzing phospholipase c, that phosphatidylcholine breakdown mimics th ...19911845988
purification and characterization of two-subunit cytochrome aa3 from bacillus cereus.cytochrome c-oxidase type aa3 (ec 1.9.3.1) was purified to homogeneity from vegetative bacillus cereus by ion-exchange and hydroxylapatite chromatography in the presence of triton x-100. gel filtration analysis suggested a dimeric structure apparently 172 kda in size; however, only a monomer of 81 kda was detected when analysed by non-denaturing gel electrophoresis. denaturing gel electrophoresis analysis of the protein showed the presence of two subunits (51 and 30 kda). atomic absorption and v ...19911849607
the aeromonas hydrophila cpha gene: molecular heterogeneity among class b metallo-beta-lactamases.an aeromonas hydrophila gene, named cpha, coding for a carbapenem-hydrolyzing metallo-beta-lactamase, was cloned in escherichia coli by screening an aeromonas genomic library for clones able to grow on imipenem-containing medium. from sequencing data, the cloned cpha gene appeared able to code for a polypeptide of 254 amino acids whose sequence includes a potential n-terminal leader sequence for targeting the protein to the periplasmic space. these data were in agreement with the molecular mass ...19911856163
travelers' diarrhea among united states military personnel during joint american-egyptian armed forces exercises in cairo, egypt.a study was conducted of travelers' diarrhea in a united states military population on deployment in cairo, egypt, during july and august 1987. acute diarrhea requiring medical attention developed in 183 (4%) of 4,500 troops. a possible etiologic agent was identified in 49% of all diarrhea cases. enteric pathogens associated with cases of diarrhea included: enterotoxigenic escherichia coli (17% st-producers, 13% lt-producers, and 3% lt/st-producers); shigella (9%); campylobacter spp. (2%); salmo ...19911900113
production of 2-methyl-4[3h]-quinazolinone, an inhibitor of poly(adp-ribose) synthetase, by bacterium. 19911900501
periodic acid-schiff-positive organisms in primary cutaneous bacillus cereus infection. case report and an investigation of the periodic acid-schiff staining properties of bacteria.primary cutaneous bacillus cereus infection frequently presents as a single necrotic bulla on the extremity of an immunocompromised patient. in lesional biopsy specimens and smears, the large gram-positive rods of b cereus may be mistaken for clostridium species. this is a potentially serious error, as bacillus species are resistant to penicillin and other beta-lactam antibiotics. we studied a case in which large periodic acid-schiff-staining organisms were seen in the biopsy specimen from a nec ...19911900984
in vitro susceptibilities of ocular bacillus cereus isolates to clindamycin, gentamicin, and vancomycin alone or in combination.a broth dilution assay was used to determine the in vitro susceptibilities of 10 ocular isolates of bacillus cereus to clindamycin, gentamicin, and vancomycin both alone and in combination. the checkerboard technique was used to determine fractional inhibitory and bactericidal concentration indices for combinations of clindamycin-gentamicin and vancomycin-gentamicin.19911901697
structural and stereospecific requirements for the nucleoside-triggered germination of bacillus cereus spores.a selection of adenosine analogues was tested for their ability to trigger germination of bacillus cereus ncib 8122 spores. the germination-inducing activity was governed by the structural properties of the sugar rather than the base moieties of the nucleosides. among the sugar-modified analogues, only those containing a 2'-deoxy-d-ribose moiety promoted spore germination. requirements for a specific molecular structure of the base were not clearly identified, although the highest activity was o ...19911901905
further observations on enhancement of the infectivity of fusobacterium necrophorum by other bacteria.it had already been shown with a single virulent strain (a42) of fusobacterium necrophorum that suspension of the fusobacteria in sub-lethal doses of broth cultures of other bacteria reduced the minimum infective dose (greater than 10(6) organisms) for mice by subcutaneous inoculation, sometimes to less than 10 organisms. the present study extended the known range of bacteria with strong infectivity-enhancing properties to include bacillus cereus, klebsiella oxytoca and staphylococcus aureus; an ...19911902184
characterization of the components of hemolysin bl from bacillus cereus.previously we described the partial purification of a novel hemolysin from bacillus cereus and showed that hemolytic activity required the combined action of at least two components, called b and l to signify their cell-binding and cell-lytic roles in this activity. on further purification, as described in the present article, a combination of anion-exchange chromatography and polyacrylamide gel electrophoresis separated three proteins, b, l1, and l2 (35, 36, and 45 kda, respectively). individua ...19911902196
hemolytic and sphingomyelinase activities of clostridium perfringens alpha-toxin are dependent on a domain homologous to that of an enzyme from the human arachidonic acid pathway.the n-terminal domain of clostridium perfringens alpha-toxin, homologous with the nontoxic phospholipase c of bacillus cereus, was expressed in escherichia coli and shown to retain all of the phosphatidylcholine hydrolyzing activity of the alpha-toxin, but not the sphingomyelinase, hemolytic, or lethal activities. the c-terminal domain of alpha-toxin showed sequence and predicted structural homologies with the n-terminal region of arachidonate 5-lipoxygenase, an enzyme from the human arachidonic ...19911902199
mureidomycin a, a new inhibitor of bacterial peptidoglycan synthesis.mureidomycin a (mrd), a novel peptidylnucleoside antibiotic with antipseudomonal activity, inhibited not only peptidoglycan synthesis but also lipid-intermediate formation from udp-n-acetylmuramyl (murnac)-pentapeptide and udp-n-acetylglucosamine in an in vitro peptidoglycan-synthesizing system, using ether-treated cells of pseudomonas aeruginosa. both types of inhibition by mrd disappeared when udp-murnac-pentapeptide was preincubated with ether-treated cells. moreover, mrd completely inhibited ...19911902646
inhibition of high-affinity gamma-aminobutyric acid uptake in primary astrocyte cultures by phorbol esters and phospholipase c.the effects of phorbol 12-myristate 13-acetate (pma), a potent activator of protein kinase c (pkc), on high-affinity na(+)-dependent gamma-aminobutyric acid (gaba) uptake were investigated in primary cultures of neurons and glial cells from rat brain cortex. incubation of glial cells with pma led to concentration- and time-dependent decreases in the gaba transport in glial cells. this effect could be completely suppressed by addition of the pkc inhibitor h7. the pma effects could be mimicked by ...19911902665
bacillus cereus: a significant pathogen in postoperative and post-traumatic wounds on orthopaedic wards.over a period of 19 months, bacillus species, most frequently b. cereus, were isolated from postoperative and post-traumatic wounds in 24 patients on orthopaedic wards in a swedish hospital. clinical signs of infection were found in all but 3 patients: in 9 patients, mild signs with increased secretion from the wounds; in 10 patients, moderate signs with fever and/or significant local reaction around the wounds; and in 2 patients, severe signs with necrotic infections. bacillus spp. were isolate ...19911902983
successful treatment of bacillus cereus infection with ciprofloxacin.bacillus cereus is rarely a pulmonary pathogen but may cause pneumonia in immunocompromised patients. a patient with bronchiectasis and no recognisable immunodeficiency had this organism isolated during two infective exacerbations, once from respiratory secretions and once by blood culture. ciprofloxacin treatment was effective on both occasions.19911902995
effects of salt concentration on bacterial growth on plates with gradients of ph and temperature.a temperature gradient device has been used to produce ph-temperature gradient plates. the response of salmonella typhimurium and bacillus cereus to continuous gradients of ph and temperature has been mapped. a third variable, sodium chloride, was investigated by incorporating it at different concentrations in a series of gradient plates. both organisms exhibited an optimum temperature and ph range for maximum salt tolerance.19911903754
envelope alteration of escherichia coli hb101 carrying peap31 caused by kil peptide and its involvement in the extracellular release of periplasmic penicillinase from an alkaliphilic bacillus.the plasmid peap31 contains the colicin e1 kil gene. peptidoglycan and outer-membrane components (lipopoly-saccharide, proteins and phosphatidylethanolamine) decreased concurrently in the envelope fraction from escherichia coli hb101 carrying peap31 during the stationary phase of growth. at almost the same time. d-alanine residues in peptidoglycan decreased. the kil peptide is suggested to affect, directly or indirectly, the turnover of peptidoglycan in stationary phase and, as a result, to caus ...19911903920
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