Bacillus

Mol Immunol, 1990 Oct, 27(10), 965 - 71
Polyvalent interaction of antibodies with bacterial cells; Karulin AYu et al.; We have studied the physical-chemical characteristics of the interaction of peroxidase-labelled rabbit antibodies with Bacillus sp . bacterial cells . The antibodies are able to bind bivalently with two antigen sites on the bacterial cells with the formation of intramolecular "cyclic" complexes . A kinetic model is proposed suggesting the existence of monovalent and bivalent cell surface antigens . The equilibrium constant of the bivalent IgG binding to the bacterial cell is by two orders of magnitude higher as compared to monovalent Fab fragments . The intramolecular reaction between the free active site of the monovalently bound antibody and a free antigen site on the cell surface is the rate limiting step of the polyvalent interaction . Formation of the cyclic complexes seems to be accompanied by essential tension of bonds and deformation of the IgG molecule . Agglutination of bacterial cells was also studied . The cell agglomerate size dependence on the antibody concn has a threshold . Agglutination proceeds under conditions where the antigen-antibody binding on the cell surface is far from equilibrium.

Surgery, 1990 Oct, 108(4), 619 - 27; discussion 627-8
Short-term tamoxifen plus chemotherapy: superior results in node-positive breast cancer; Crowe JP Jr et al.; Three hundred eleven patients with node-positive breast cancer were randomized to one of three adjuvant treatments: cyclophosphamide (Cytoxan), methotrexate, and 5-fluorouracil; all of the above with tamoxifen citrate; or all of the above with tamoxifen and bacillus Calmette-Guerin vaccination . Local therapy for all patients was a modified radical mastectomy . Estrogen receptors were measured on all primary tumors . Patients were stratified by the number of positive nodes (one to three nodes and more than three nodes) and estrogen-receptor value (less than 3 femtomole/mg and greater than or equal to 3 femtomole/mg) . Follow-up is available, with a mean of 9.1 and maximum of 14.2 years . In this study the efficacy of short-term tamoxifen is apparent over that of chemoimmunotherapy alone and continues to be significant with prolonged follow-up . The addition of tamoxifen to chemoimmunotherapy significantly prolonged disease-free survival among patients with estrogen receptor-positive tumors who were postmenopausal, who had larger tumors (greater than 3 cm), or who had more extensive axillary node involvement (more than three nodes) . Tamoxifen improved overall survival for patients with estrogen receptor-positive tumors larger than 3 cm . The addition of bacillus Calmette-Guerin Cytoxan, methotrexate, 5-fluorouracil, and tamoxifen did not significantly alter disease-free or overall survival.

J Bacteriol, 1990 Oct, 172(10), 6156 - 9
Presence of N6-methyladenine in GATC sequences of Bacillus popilliae and Bacillus lentimorbus KLN2; Dingman DW; Nine strains of Bacillus popilliae and Bacillus lentimorbus KLN2 contain N6-methyladenine in GATC sequences, as determined by using the restriction enzymes MboI and DpnI . Among eight other Bacillus species examined, all, except one strain of Bacillus brevis (ATCC 9999), lacked adenine methylation in GATC . A methylase with Escherichia coli dcm site specificity was not present in any of the Bacillus species studied.

J Biochem (Tokyo), 1990 Oct, 108(4), 583 - 7
Purification and characterization of catalase from a facultative alkalophilic Bacillus; Yumoto I et al.; Catalase was purified to an electrophoretically homogeneous state from the facultative alkalophilic bacterium, Bacillus YN-2000, and some of its properties were studied . Its molecular weight was 282,000 and its molecule was composed of four identical subunits . The enzyme contained two protoheme molecules per tetramer . The enzyme showed an absorption spectrum of typical high-spin ferric heme with a peak at 406 nm in the oxidized form and peaks at 440, 559, and 592 nm in the reduced form . In contrast to the typical catalases, the enzyme was reduced with sodium dithionite, like peroxidases . The enzyme showed an appreciable peroxidase activity in addition to high catalase activity . The amino acid composition of Bacillus YN-2000 catalase was very similar to those of catalase from Neurospora crassa and peroxidase from Halobacterium halobium . The catalase content in the soluble fraction from the bacterium was higher with the cells grown at pH 10 than with the cells grown at lower pHs (pH 7-9).

Protein Eng, 1990 Oct, 4(1), 65 - 7
Engineering a heavy atom derivative for the X-ray structure analysis of cyclodextrin glycosyltransferase; Klein C et al.; Based on a preliminary structural model of cyclodextrin glycosyltransferase from Bacillus circulans (EC 2.4.1.19), Ser428 and Ser475 of the enzyme were mutated to cysteines in order to produce suitable heavy atom derivatives . Mutant Ser475----Cys could not be expressed as protein . Mutant Ser428----Cys was expressed in Escherichia coli and purified . It crystallized isomorphously and gave rise to a mercury derivative that improved the electron density map . The structural results show that the new mercury-binding site is in a pocket at the protein surface.

Arch Biol Med Exp (Santiago), 1990 Oct, 23(2), 101 - 12
Evaluation of soil microorganisms with inhibitory activity against Rhizoctonia solani causal agent of the damping-off of canola; Ciampi L et al.; Pre- and post-emergence damping-off of canola seedlings caused by Rhizoctonia solani is a serious disease in Western Canada . Other fungi such as Fusarium spp . and Pythium spp . are also related to seedling damping-off . To-day, the search of soil bacteria is becoming a tool to use microorganisms as potential biocontrol agents for several plant diseases . The purpose of this research was to detect bacteria to biologically control R . solani, Pythium spp., and Fusarium spp . Soil samples were collected throughout Alberta during 1987 to isolate bacteria . Canola seedlings were also used to obtain bacteria from the same samples . Plant pathogenic fungi were tested to detect the antagonistic activity of the isolates . Tests were made with coated canola seeds, amendments and fresh of freeze-dried cells . Three hundred forty-one bacterial cultures were isolated . Only 16 inhibited fungal growth: 7 showed the same effects against R . solani and 9 showed uneven effects . Some isolates showed a weak action to Pythium spp . and Fusarium spp . Three isolates showed inhibitory effect on R . solani and Pythium spp . Isolate F1 improved by about 50% the germination of canola seeds in inoculated pots when compared with the inoculated control . Coated seeds had low germination and emergence was below the inoculated control . The emergence of canola seedlings was very much improved when isolate 147 was delivered as an amendment in inoculated pots . Identification showed that 3 bacterial belonged to Bacillus spp., 4 to green fluorescent Pseudomonas spp . and 2 were Streptomyces spp.

Rev Med Chir Soc Med Nat Iasi, 1990 Oct-Dec, 94(3-4), 491 - 3
{Risk factors in chronic posttubercular cor pulmonale in the elderly}; Nicola V et al.; From the analysis of 866 aged patients with post tuberculosis sequels admitted to the Tuberculosis Hospital at Constanta in the interval 1979-1988, a high percentage of the post-tuberculosis respiratory syndromes (41.8%) was recorded . Post-tuberculosis chronic cor pulmonale was in 62 patients (17%) . The recent onset or reactivation of the bacillary process seem to represent important risk factors for such patients . The old age and other associated diseases usually restrict the therapeutical possibilities and, consequently, delay the cure.

J Appl Bacteriol, 1990 Oct, 69(4), 557 - 62
Forces involved in adhesion of Bacillus cereus spores to solid surfaces under different environmental conditions; Husmark U et al.; The adhesion of Bacillus cereus spores (NCTC 2599) to hydrophobic and hydrophilic glass surfaces was studied when environmental conditions were varied . The spores were exposed in media of different polarities as well as different pH and ionic concentrations . With increasing ethanol concentrations, the polarity of the medium was decreased and the predominant force of attraction was found to be hydrophobic . The spore surface was uncharged at a pH around 3, at which value the spore was most adhesive to both hydrophobic and hydrophilic glass . This could be attributable to the absence of electrostatic repulsion . An increased ionic concentration of the bulk increased the degree of adhesion especially to the hydrophilic surfaces . This indicates the suppression of a solvation barrier at high ionic concentrations, when the polymers of the spore surface become dehydrated.

FEMS Microbiol Lett, 1990 Oct, 60(1-2), 123 - 6
Plasmid transformation of Bacillus cereus on cellophane membranes; Sabelnikov AG et al.; A simple approach to test the ability of bacteria to undergo natural genetic transformation is suggested . The basic feature of the approach is the cultivation of bacterial cells in the presence of exogenous (plasmid) DNA on cellophane membranes placed successively on nutrient and selective agar . Using this approach the ability of Bacillus cereus for "natural" genetic transformation was detected . Transformation frequencies varied from 10(-8) to 10(-6).

Panminerva Med, 1990 Oct-Dec, 32(4), 199 - 201
A case of subprimary suprarenal tuberculosis; Bonanni G et al.; In spite of the current availability of new and more up-to-date methods of prevention, diagnosis and therapy, specific forms in general and the ability to perform rapid, efficacious and resolutory operations, a number of cases of subprimary or secondary tuberculosis still exist which are most probably the result of delayed diagnosis, contagious episodes or the failure to adhere to recommended treatment protocols and, often, less than optimum medical therapy . A rare case of adrenal tuberculosis diagnosed in a patient hospitalized for left renal calculosis is reported . Routine tests indicated a specific form of diagnosis, and during surgery for left renal calculosis the patient underwent left adrenalectomy . Histological tests confirmed the initial diagnosis . For 180 days after the operation the patient received pharmacological treatment directed specifically against Koch's bacillus.

Chem Pharm Bull (Tokyo), 1990 Oct, 38(10), 2887 - 9
Inhibitory effect of bisbenzylisoquinoline alkaloids on the quick death of mice treated with BCG/LPS; Kondo Y et al.; Three bisbenzylisoqinoline alkaloids, chondocurine (Chon), berbamine (Ber), and cycleanine (Cyc) were tested for their protective effect on the quick death of mice primed with bacillus Calmette Guerin (BCG) and elicited with lipopolysaccharide (LPS) . Seven-consecutive treatments with Chon or Cyc at a dose of 10 mg/kg following BCG priming resulted in significant improvement in the survival rate . A single dose of Chon also protected the BCG/LPS-treated mice from death if it was given immediately after, not 4 h after, LPS elicitation . These data show that bisbenzylisoquinoline alkaloids can protect mice from the lethal toxicity induced by the BCG/LPS combination treatment by inhibiting the priming with BCG or the elicitation with LPS.

Ecotoxicol Environ Saf, 1990 Oct, 20(2), 211 - 21
Employment of CEPEX enclosures for monitoring toxicity of Hg and Zn on in situ structural and functional characteristics of algal communities of River Ganga in Varanasi, India; Rai LC et al.; Effects of Hg and Zn on in situ nitrogen fixation, autotrophic index, pigment diversity, 14CO2 uptake, and change in algal community structure of Ganges water have been studied for the first time using CEPEX chambers in aquatic ecosystem of India . A concentration-dependent decrease in in situ nitrogenase activity of Ganges water with Hg and Zn has been noticed . No ethylene production was observed at 0.8 microgram/ml of Hg . However, an increase in the autotrophic index was observed in CEPEX enclosures treated with Hg and Zn . The AI value was maximum at 0.8 microgram/ml Hg after an incubation of 15 days . An increase in pigment diversity also followed the pattern of AI with the test metals used . Inhibition of 14CO2 uptake of phytoplankton of Ganges water was maximum at 0.8 microgram/ml Hg (79%) followed by Zn (69%) . Carbon fixation showed an increase for 1 hr, after which no appreciable change was noticed . Maximum inhibition of algal number was observed at 0.8 microgram/ml Hg followed by 8.0 micrograms/ml of Zn in the CEPEX chamber . Members of Chlorophyceae showed more tolerance than Cyanophyceae and Bacillariophyceae . The filamentous forms were more tolerant to Hg and Zn . In contrast, unicellular forms were more sensitive to Hg . The test of significance (ANOVA) showed that metal-induced variations in pigment diversity, the autotrophic index, and the 14CO2 uptake were highly significant (P less than 0.001).

Am J Trop Med Hyg, 1990 Oct, 43(4), 373 - 9
Bb65, a major immunoreactive protein of Bartonella bacilliformis; Knobloch J et al.; A 65 kDa protein (Bb65) has been identified as one of the major specific antigens of Bartonella bacilliformis, the causative agent of bartonellosis which is a bacterial infectious disease of inhabitants of the Andes . The gene encoding this antigen (7B2) was isolated from an expression library made directly from randomly generated fragments of B . bacilliformis genomic DNA using Bartonella antibodies raised in rabbits and sera of bartonellosis patients . The Bartonella 7B2 gene was expressed in Escherichia coli and the recombinant Bb65 protein was purified by column chromatography . Using polyclonal antibodies raised in rabbits, the antigen was shown to be present in all of 13 B . bacilliformis isolates from different Peruvian regions . Immune electron microscopy demonstrated the probable cytoplasmatic localization of Bb65 . When applied to enzyme immunoassays, Bb65 sensitively and specifically bound to IgG antibody of sera of bartonellosis patients, convalescents, and immunes from various Peruvian regions . IgM antibody was not recognized by Bb65, neither was IgG antibody circulating during the first 2 weeks of illness . The amino-terminal amino acid sequence of Bb65 was 53% homologous to the 65 kDa heat shock protein of Mycobacterium tuberculosis.

Appl Microbiol Biotechnol, 1990 Oct, 34(1), 57 - 62
Alkaline serine protease produced from citric acid by Bacillus alcalophilus subsp . halodurans KP 1239; Takii Y et al.; Maximum production of alkaline serine protease by Bacillus alcalophilus subsp . halodurans KP 1239 was achieved after 24 h cultivation, at an initial pH of 7.6, on a medium containing 1.0% sodium citrate, 0.3% yeast extract, and 0.3% KH2PO4 . The enzyme was purified to crystalline form from culture broth . The enzyme was most active at 60 degrees C and at pH 11.5 . The molecular weight, isoelectric point and sedimentation coefficient in water at 20 degrees C were estimated as 29,000, 8.8 and 3.3S, respectively . The N-terminal amino acid sequence was Ala-Gln-Ser-Val-Pro-Trp-Gly-Ile-Ser-Arg-Val-Gln-Ala-Pro-Ala-Ala- His-Asn-Arg-Gly- . The enzyme shared its antigenic determinants with B . alcalophilus ATCC 21522 serine protease, but not with the subtilisins Carlsberg and BPN'.

Biotechnology (N Y), 1990 Oct, 8(10), 939 - 43
Insect resistant cotton plants; Perlak FJ et al.; We have expressed truncated forms of the insect control protein genes of Bacillus thuringiensis var . kurstaki HD-1(cryIA(b) and HD-73 (cryIA(c) in cotton plants at levels that provided effective control of agronomically important lepidopteran insect pests . Total protection from insect damage of leaf tissue from these plants was observed in laboratory assays when tested with two lepidopteran insects, an insect relatively sensitive to the B.t.k . insect control protein, Trichoplusia ni (cabbage looper) and an insect that is 100 fold less sensitive, Spodoptera exigua (beet armyworm) . Whole plants, assayed under conditions of high insect pressure with Heliothis zea (cotton bollworm) showed effective square and boll protection . Immunological analysis of the cotton plants indicated that the insect control protein represented 0.05% to 0.1% of the total soluble protein . We view these results as a major step towards the agricultural use of genetically modified plants with insect resistance in this valuable, high acreage crop.

Biochemistry, 1990 Sep 18, 29(37), 8587 - 91
Designs for a broad substrate specificity keto acid dehydrogenase; Wilks HM et al.; Variations have been made to the structure of the nicotinamide adenine dinucleotide (NAD) dependent L-lactate dehydrogenase from Bacillus stearothermophilus at regions of the enzyme that we believe determine specificity toward different alpha-hydroxy acids (RCHOHCOO-, R = CH3, C2H5, etc.) . Two regions of LDH that border the active site (but are not involved in the catalytic reaction) were altered in order to accommodate substrates with hydrophobic side chains larger than that of the naturally preferred substrate, pyruvate (R = CH3) . The mutations 102-105GlnLysPro----MetValSer and 236-237AlaAla----GlyGly were made to increase the tolerance for large hydrophobic substrate side chains . The triple and double mutants alone gave little improvement for branched-chain-substituted pyruvates . The five changes together produced a broader substrate specificity alpha-hydroxy acid dehydrogenase, with a 55-fold improved kcat for alpha-ketoisocaproate to a value about 1/14 that of the native enzyme for pyruvate . Rational protein engineering enabled coupled changes in enzyme structure to be obtained with greater probability of success than random mutagenesis.

J Biol Chem, 1990 Sep 15, 265(26), 15481 - 8
Use of amber suppressors to investigate the thermostability of Bacillus licheniformis alpha-amylase . Amino acid replacements at 6 histidine residues reveal a critical position at His-133; Declerck N et al.; A set of 12 Escherichia coli suppressor tRNAs, inserting different amino acids in response to an amber codon, has been used to create rapidly numerous protein variants of a thermostable amylase; by site-directed mutagenesis, amber mutations were first introduced into Bacillus licheniformis alpha-amylase gene at position His35, His133, His247, His293, His406, or His450; genes carrying one or two amber mutations were then expressed in the different suppressor strains, generating over 100 amylase variants with predicted amino acid changes that could be tested for thermostability . Within the detection limits of the assays, amino acid replacements at five histidine positions had no significant effect . In contrast, suppressed variants substituted at residue His133 clearly exhibited modified thermostability and could be either less stable or more stable than the wild-type amylase, depending on the amino acid inserted at this position; comparison of the variants indicates that the hydrophobicity of the substituting residue is an important but not a determinant factor of stabilization . The effect of the most stabilizing and destabilizing amino acid substitutions, His133 to Tyr and to Pro, respectively, were confirmed by introducing the corresponding missense mutations into the gene sequence . The advantages and limits of informational suppression in protein stability studies are discussed as well as structural features involved in the thermostability of B . licheniformis alpha-amylase.

Eur J Biochem, 1990 Sep 11, 192(2), 337 - 45
Structural features of the lipopeptidophosphoglycan from Trypanosoma cruzi common with the glycophosphatidylinositol anchors; de Lederkremer RM et al.; The lipopeptidophosphoglycan (LPPG) from Trypanosoma cruzi, a major constituent of the plasma membrane of epimastigote forms, has been now extracted with butanol/water from delipidated cells and purified by hydrophobic chromatography . We have found that the LPPG undergoes two reactions, characteristic of the glycosylphosphatidylinositol anchors: (a) cleavage of the ceramide by phosphatidylinositol-specific phospholipase C (PtdIns-specific phospholipase C) from Bacillus thuringiensis, (b) nitrous acid deamination of the non-N-acylated glucosamine . Palmitoylsphinganine, palmitoylsphingosine, lignoceroylsphinganine and, as minor components, the stearoylceramides were identified by gas liquid chromatography/mass spectrometry . The presence of cross reacting determinant (CRD) epitopes in the glycophosphoinositol released by PtdIns-specific phospholipase C was investigated by direct and inhibition ELISA . A sample of glycophosphoinositol containing 5 micrograms carbohydrate caused 60% inhibition of the binding of anti-CRD antibodies raised against the soluble form of variant surface glycoprotein.

Biochim Biophys Acta, 1990 Sep 10, 1087(1), 73 - 9
Expression of the copy DNA for human A4 and B4 L-lactate dehydrogenases in Escherichia coli; Barstow DA et al.; The human LDH-A and LDH-B cDNAs, containing the coding regions for the L-lactate dehydrogenase A4 (M) and B4 (H) polypeptides respectively have been cloned into Escherichia coli to place the cDNAs under the control of hybrid E . coli/Bacillus stearothermophilus transcriptional and translational signals . Human A4- and B4-isoenzymes are produced in E . coli cells harbouring the expression plasmids pHLDHA22 and pHLDHB10 at levels of 6.5 and 1.5% of the soluble protein of the cell, respectively . The tac promoter of these vectors was not induced by isopropyl beta-D-thiogalactopyranoside . The A4 and B4 human isoenzymes synthesized in E . coli were purified to homogeneity and show the same properties as isoenzymes isolated from human tissue . The amino acid sequences of 12 N-terminal residues of the human isoenzymes synthesized in E . coli were determined to be identical to those deduced from the DNA sequence of the cloned cDNAs except that the N-terminal methionine was absent from both . However, in contrast to LDH made in human cells, acetylation of the N-terminal alanine does not take place in E . coli cells.

J Biol Chem, 1990 Sep 5, 265(25), 15034 - 9
Rabbit small intestinal trehalase . Purification, cDNA cloning, expression, and verification of glycosylphosphatidylinositol anchoring; Ruf J et al.; alpha,alpha-Trehalase (EC 3.2.1.28), an intrinsic protein of intestinal brush-border membranes, was purified to homogeneity from rabbits . Partial amino acid sequences were determined . Two degenerate oligonucleotides based on the sequence of a CNBr peptide were employed in a polymerase chain reaction to amplify a 71-base pair fragment of trehalase DNA with rabbit intestine cDNA as a starting template . This fragment was used as a hybridization probe to isolate full length trehalase clones from a rabbit intestine cDNA bank . Sequence analysis revealed that trehalase comprises 578 amino acids, contains at the amino terminus a typical cleavable signal sequence, at the carboxyl terminus a rather hydrophobic region typical of proteins anchored via glycosylphosphatidylinositol, and four potential N-glycosylation sites . Trehalase has no sequence homologies with other sequenced brush-border glycosidases . Northern blot analysis revealed a 1.9-kilobase trehalase mRNA in small intestine and kidney, smaller amounts in liver, and none in lung . Southern blot analysis indicated the gene has a length of 20 kilobase pairs or less . Injection into Xenopus laevis oocytes of mRNA synthesized in vitro from a trehalase template resulted in the expression of trehalase activity several hundredfold above background . The trehalase activity was membrane-bound and could be solubilized upon digestion with phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis . This strongly suggests that rabbit small intestinal trehalase is anchored via glycosylphosphatidylinositol also when expressed in X . laevis oocytes.

Biochemistry, 1990 Sep 4, 29(35), 8056 - 62
Phosphatidylinositol-specific phospholipase C from Bacillus cereus combines intrinsic phosphotransferase and cyclic phosphodiesterase activities: a 31P NMR study; Volwerk JJ et al.; The inositol phosphate products formed during the cleavage of phosphatidylinositol by phosphatidylinositol-specific phospholipase C from Bacillus cereus were analyzed by 31P NMR . 31P NMR spectroscopy can distinguish between the inositol phosphate species and phosphatidylinositol . Chemical shift values (with reference to phosphoric acid) observed are 0.41, 3.62, 4.45, and 16.30 ppm for phosphatidylinositol, myo-inositol 1-monophosphate, myo-inositol 2-monophosphate, and myo-inositol 1,2-cyclic monophosphate, respectively . It is shown that under a variety of experimental conditions this phospholipase C cleaves phosphatidylinositol via an intramolecular phosphotransfer reaction producing diacylglycerol and D-myo-inositol 1,2-cyclic monophosphate . We also report the new and unexpected observation that the phosphatidylinositol-specific phospholipase C from B . cereus is able to hydrolyze the inositol cyclic phosphate to form D-myo-inositol 1-monophosphate . The enzyme, therefore, possesses phosphotransferase and cyclic phosphodiesterase activities . The second reaction requires thousandfold higher enzyme concentrations to be observed by 31P NMR . This reaction was shown to be regiospecific in that only the 1-phosphate was produced and stereospecific in that only D-myo-inositol 1,2-cyclic monophosphate was hydrolyzed . Inhibition with a monoclonal antibody specific for the B . cereus phospholipase C showed that the cyclic phosphodiesterase activity is intrinsic to the bacterial enzyme . We propose a two-step mechanism for the phosphatidyl-inositol-specific phospholipase C from B . cereus involving sequential phosphotransferase and cyclic phosphodiesterase activities . This mechanism bears a resemblance to the well-known two-step mechanism of pancreatic ribonuclease, RNase A.

Urology, 1990 Sep, 36(3), 222 - 5
Maintenance intravesical bacillus calmette-guerin for superficial transitional cell carcinoma of bladder . A retrospective study; Schwalb DM et al.; Eighteen patients with recurrent and/or multifocal superficial transitional cell carcinoma of the bladder who were rendered tumor-free by transurethral resection and were then treated with either a single or second six-week course of induction Bacillus Calmette-Guerin (BCG) therapy, followed by maintenance therapy, were retrospectively reviewed . A 73 percent complete response rate was achieved in those patients treated prophylactically, while a 70 percent complete response rate was observed in patients treated for carcinoma in situ (CIS) with an average follow-up of twenty-nine months . Maintenance therapy may be warranted in those patients able to tolerate it without significant side effects.

Antimicrob Agents Chemother, 1990 Sep, 34(9), 1725 - 32
Nucleotide sequence and phylogeny of SHV-2 beta-lactamase; Huletsky A et al.; We determined the nucleotide sequence of the blaSHV-2(pBP60-1) gene from Klebsiella ozaenae which confers resistance to broad-spectrum cephalosporins . The structural gene encodes a polypeptide product of 286 amino acids, and the estimated molecular weight of the mature protein is 28,900 . Amino acid sequence comparison of the SHV-2pBP60-1 enzyme with all known class A beta-lactamases and homology studies showed that the residues were highly conserved . Furthermore, SHV-2pBP60-1 was clearly related to SHV-1, LEN-1, and OHIO-1 . The SHV-2pBP60-1 enzyme differed from SHV-1 isolated from Klebsiella pneumoniae by seven amino acid substitutions . One of these substitutions, the Gly----Ser substitution at position 234, is probably a key region for the novel activity of cefotaxime hydrolysis . A phylogenetic tree was constructed by using all class A beta-lactamases of known sequences by a progressive alignment method . The data suggested that the beta-lactamases of gram-positive Streptomyces, Staphylococcus, and Bacillus species appeared early in evolution, followed by the PSE and CARB enzymes of Pseudomonas species and, more recently, by the SHV-type and TEM-type enzymes found in enteric bacteria . Larger evolutionary distances separated clusters of the gram-positive beta-lactamases than separated clusters of the gram-negative enzymes . Results of this phylogenetic study suggested that extended-spectrum enzymes are recent derivatives that are selected by the use of new cephalosporins.

Trans R Soc Trop Med Hyg, 1990 Sep-Oct, 84(5), 739 - 44
Tuberculosis in developing countries and methods for its control; Rodrigues LC et al.; Tuberculosis is a major cause of morbidity and mortality in developing countries . It is estimated that one-fifth of the world population is infected, 12-16 million people have the disease, every year 6-8 million develop tuberculosis and 2-3 million die from it . Four methods for the prevention of tuberculosis are available: improvement of socio-economic conditions, case-finding and treatment, chemoprophylaxis, and vaccination . Each of these methods is examined in relation to a model of the chain of transmission and development of disease . Improvement of socio-economic conditions, responsible for the decline of tuberculosis in the developed world, must be seen as a long-term solution . Case-finding and treatment is the only method expected to have an important short-term impact on transmission . A summary of the results of 35 studies shows the estimated efficacy of bacillus Calmette-Guerin (BCG) vaccination ranging from 96% to none . Follow-up results from 10 controlled trials are consistent with waning of BCG protective efficacy with time since vaccination . Chemoprophylaxis and vaccination are expected to protect the individual but not to have a significant short-term impact on transmission.

J Biochem (Tokyo), 1990 Sep, 108(3), 379 - 81
Crystallization and preliminary crystallographic study of bacterial alpha-amylases; Suzuki A et al.; Crystallization of bacterial alpha-amylases has been achieved by the hanging-drop vapor diffusion method . The crystals of Bacillus licheniformis and B . licheniformis 584 amylases are isomorphous to each other . The crystals of B . licheniformis amylase belong to the tetragonal system, space group P4(2)2(1)2 with cell dimensions of a = 119.3 and c = 85.4 A . The asymmetric unit contains one molecule of amylase, with a volume per molecular mass, Vm, of 2.75 A3/Da . The crystals of B . licheniformis and B . licheniformis 584 amylases diffract beyond 2.5 A resolution and are suitable for X-ray diffraction analysis . The crystals of B . amyloliquefaciens amylase are orthorhombic, and have space group C222(1), with cell dimensions of a = 154, b = 298, and c = 90 A . The asymmetric unit contains three to five molecules . In the crystallization of B . licheniformis and B . licheniformis 584 amylases, the addition of EDTA was indispensable to obtain large single crystals, while it had an adverse effect on the crystallization of B . amyloliquefaciens amylase, producing a large amount of small crystals.

J Invertebr Pathol, 1990 Sep, 56(2), 258 - 66
Specificity and efficacy of purified Bacillus thuringiensis proteins against agronomically important insects; MacIntosh SC et al.; The host range and relative efficacy of three purified Bacillus thuringiensis insect control proteins were determined against 17 different agronomically important insects representing five orders and one species of mite . The three B . thuringiensis proteins were single gene products from B . thuringiensis ssp . kurstaki HD-1 (CryIA(b)) and HD-73 (CryIA(c)), both lepidopteran-specific proteins, and B . thuringiensis ssp . tenebrionis (CryIIIA), a coleopteran-specific protein . Seven insects showed sensitivity to both B . thuringiensis ssp . kurstaki proteins, whereas only 1 of the 18 insects was sensitive to B . thuringiensis ssp . tenebrionis protein . The level of B . thuringiensis ssp . kurstaki protein required for 50% mortality (LC50) varied by 2000-fold for these 7 insects . A larval growth inhibition assay was developed to determine the amount of B . thuringiensis ssp . kurstaki protein required to inhibit larval growth by 50% (EC50) . This extremely sensitive assay enabled detection of B . thuringiensis ssp . kurstaki HD-73 levels as low as 1 ng/ml.

J Invertebr Pathol, 1990 Sep, 56(2), 233 - 6
Activation and germination of Bacillus thuringiensis spores in Manduca sexta larval gut fluid; Wilson GR et al.; Incubation of Bacillus thuringiensis HD-1 spores in the larval gut fluid of Manduca sexta (tobacco hornworm) resulted in increased viable counts, conversion to phase-dark spores, and a loss of absorbance in spore suspensions, indicative of spore germination . Heat-activated and untreated spores incubated in water did not exhibit these changes . Only when spores were heat activated and incubated in germinants L-alanine and adenosine did changes in the spores approximate those observed in gut fluid . These data suggest that M . sexta larval gut fluid induces the activation and germination of B . thuringiensis spores.

J Invertebr Pathol, 1990 Sep, 56(2), 143 - 9
Selective inhibition of cecropin-like activity of insect immune blood by protease from American foulbrood scales; Jarosz J et al.; Bioassays of American foulbrood larval scale filtrates have shown the presence of an immune inhibitor with a specific activity of proteases that selectively destroy cecropin-like activity in insect immune hemolymphs . It was an unexpected phenomenon to find that Bacillus larvae protease(s), even at trace concentrations, totally inhibits bactericidal activity of immune blood against Escherichia coli D 31 . Thermal inactivation of a proteolytic enzyme coincides strictly with a disappearance of the ability to inhibit a cecropin-type activity in an assay system used, although the immune inhibitor was unaffected by trypsin digestion . Since neither bee larval hemolymph nor larval body homogenates possess demonstrable inhibitory action against cecropins present in hemolymph of Celerio euphorbiae and other insects, one can suggest that B . larvae proteases selectively destroy some molecules of the insect humoral immune system . The absence of cecropins in blood of immunized honeybee, Apis mellifera, larvae and a simultaneous high inhibitory activity of B . larvae protease(s) need further study on their role in pathogenesis of American foulbrood of the honeybee.

Izv Akad Nauk SSSR Biol, 1990 Sep-Oct, (5), 789 - 91
{Bacillus thuringiensis subsp . toguchini--a new subspecies of crystal-forming bacteria}; Khodyrev VP; A new subspecies of B . thuringiensis subsp . toguchini has been isolated from soil in Novosibirsk region . New subspecies forms elliptic crystals . Biologically and serologically it is different from other known subspecies.

Zentralbl Hyg Umweltmed, 1990 Sep, 190(3), 275 - 92
{Characteristics of a new bioindicator and its testing in practice assays of thermal disinfection methods}; Senkpiel K et al.; Thermolysin (EC 3.4.24.4) is a thermostable metalloproteinase isolated from Bacillus thermoproteolyticus . It has been tested for its suitability as a macromolecular bioindicator for the screening of thermal disinfection methods . Using the time- und temperature-dependent inactivation rate of the immobilized enzyme we get a measure if given parameters of disinfection have been maintained . In order to determine standard conditions of practical use, physico-chemical properties of the enzyme (pH- and temperature-optimum, Km, Vmax) were ascertained first . Then, thermal inactivation of thermolysin was analyzed for temperatures between 75 degrees and 93 degrees C and exposition time 3-20 min . This new method of testing thermal disinfection is compared with conventional physical and microbiological testing methods; in addition values of reference for selected disinfection programs were analyzed . The bioindicator proved to be stable for up to 12 weeks of storage (room temperature).

Mol Gen Genet, 1990 Sep, 223(2), 332 - 4
Expression of the blasticidin S deaminase gene (bsr) in tobacco: fungicide tolerance and a new selective marker for transgenic plants; Kamakura T et al.; Blasticidin S (BS), a fungicide of microbial origin, is used for the practical control of rice blast disease . It has broad antimicrobial activity but occasionally exhibits adverse phytotoxic effects on some dicot plants . An inactivating enzyme, BS deaminase, was discovered in the BS resistant strain, Bacillus cereus K55-S1, and the structural gene, bsr, for the enzyme has been cloned . We introduced the bsr gene into tobacco plants using the Ti plasmid vector system and demonstrated that the bsr gene conferred a BS resistant phenotype to the plants . Thus the bsr gene could be useful as a selective marker for plant transformation and provides an example for a new approach to the solution of phytotoxicity problems associated with the use of some types of fungicide.

Mt Sinai J Med, 1990 Sep, 57(4), 209 - 15
Pulmonary infection with Mycobacterium avium-intracellulare: diagnosis, clinical patterns, treatment; Teirstein AS et al.; Most physicians fail to recognize Mycobacterium avium-intracellulare (MAI) as a major pathogen for pulmonary disease among patients admitted to hospitals throughout the United States . In a review of all records of positive MAI cultures during the 10 years beginning July 1, 1979, at The Mount Sinai Hospital, New York City, we have identified 244 patients who had pulmonary disease primarily or secondarily complicated by MAI . We also identified another 243 patients as false positive for MAI infection . We classed as false positives patients who had no subsequent positive culture and whose clinical picture was and remained incompatible with MAI infection . We identified four distinct clinical patterns in the 244 patients with true positive MAI infections: (a) pulmonary nodules ("tuberculomas") indistinguishable from pulmonary neoplasms (78 patients); (b) chronic bronchitis or bronchiectasis with sputum repeatedly positive for MAI or granulomas on biopsy (58 patients, virtually all older white women); (c) cavitary lung disease and scattered pulmonary nodules mimicking M . tuberculosis infection (12 patients); (d) diffuse pulmonary infiltrations in immunocompromised hosts, primarily patients with AIDS (96 patients) . The diagnosis should be established either by surgical resection and culture of resected nodules, or by three repeated positive acid-fast bacillus cultures of sputum or fluid and tissue obtained by bronchoscopy, or by biopsy of other tissue which shows granulomas and one or more positive MAI cultures . Surgical resection is the best treatment for "solitary" MAI nodules . Multiple antituberculous drug therapy is indicated for patients with chronic infection that impairs function or causes hemoptysis . The presence of MAI in the sputum or lung aspirates of patients with AIDS usually heralds the presence of a preterminal disseminated infection.

Tubercle, 1990 Sep, 71(3), 169 - 72
Chromosome damage in untreated tuberculosis patients; Rao VV et al.; The frequency of chromosomal aberrations and sister chromatid exchanges (SCEs) were estimated in the lymphocytes of 95 untreated tuberculosis patients to evaluate the chromosomal damage with reference to bacterial invasion . The frequency of chromosomal aberrations was significantly higher in tuberculosis patients compared to controls (p less than 0.001) while the SCE frequency did not show any statistically significant differences . These findings indicate that the basic mechanism of formation of chromosomal aberrations and SCE are different and they suggest that the tubercle bacillus might be capable of inducing chromosome damage.

J Am Mosq Control Assoc, 1990 Sep, 6(3), 496 - 9
Evaluation of Bacillus sphaericus 2362 against Culex quinquefasciatus in septic ditches; Jones JW et al.; Four application rates of Bacillus sphaericus strain 2362 were tested for efficacy in septic ditches against Culex quinquefasciatus 2nd-4th instar larvae . Trials were conducted over a 2-year period . In 1987, all dosages applied to dairy effluent ditches resulted in substantial reductions after 48 h . Differences among dosages did not differ significantly (P greater than or equal to 0.05) . After 5 days, mosquito larval numbers increased with dosage rates of 0.6, 0.9 and 1.2 liters/ha . Residual control was maintained, however, at 2.4 liters/ha for 17 days . In 1988, B . sphaericus applied to domestic sewage effluent ditches at rates of 1.2 liters/ha, and 2.4 liters/ha induced greater than 79% suppression for a period of 15 days, and the 1.2 liters/ha rate induced greater than 79% suppression for a period of 20 days . A rate of 0.9 liters/ha provided good to excellent control (greater than 88%) for a period of 10 days . A rate of 0.6 liters/ha produced less than 50% suppression after 48 h.

J Am Mosq Control Assoc, 1990 Sep, 6(3), 384 - 9
Fate and persistence of Bacillus sphaericus used as a mosquito larvicide in dairy wastewater lagoons; Matanmi BA et al.; The fate and persistence of the mosquitocidal bacterium, Bacillus sphaericus, in dairy wastewater lagoons was evaluated in conjunction with trials of its larvicidal efficacy against Culex stigmatosoma . Two commercial formulations, BSP-2 (at 4.48 kg/ha) and ABG-6184 (at 2.24 kg/ha) gave about 90% reduction for up to 4 weeks, although surface water lost its insecticidal activity by 3 days posttreatment . Spores settled to the bottom within 3 days of treatment, but could be recovered in surface water after reflooding . Spore concentrations in bottom water varied widely, yet insecticidal activity remained high for from 3 days (BSP-2 at 4.48 kg/ha) to 2 weeks (ABG-6184 at 2.24 kg/ha) . Spores persisted in the mud throughout the study period . These results indicate the extended control obtained was due primarily to the ingestion of spores from bottom water and mud by larvae, which routinely inhabit the shallow areas toward the edge of the pond and browse at the pond bottom.

J Clin Microbiol, 1990 Sep, 28(9), 2114 - 6
Bacillus species pseudobacteremia traced to contaminated gloves used in collection of blood from patients with acquired immunodeficiency syndrome; York MK; Ten nonpathogenic Bacillus isolates were obtained from blood cultures collected over a 2-year period . Eight of these isolates were from patient with acquired immunodeficiency syndrome, and seven were recovered from blood cultures obtained in outpatient clinics . Five cases occurred during a 5-month period . These five cases were clinically evaluated, and the Bacillus isolates were characterized . The same Bacillus species was isolated from nonsterile gloves from the same lot worn by phlebotomists for blood collection in the outpatient clinics during this period, implicating the gloves as the cause of this pseudoepidemic . Awareness of the nonsterile nature of gloves used by laboratory personnel should be considered in the evaluation of Bacillus spp . in blood cultures.

Thorax, 1990 Sep, 45(9), 709 - 10
Pulmonary disease following intravesical BCG treatment; Kesten S et al.; Bacillus Calmette-Guerin (BCG) is an attenuated strain of Mycobacterium bovis that has been used in the treatment of malignant disease for over 20 years and for the treatment of bladder cancer since 1976 . Major complications of this treatment are infrequent . We report two cases of systemic illness with pulmonary manifestations after treatment with intravesical BCG.

Int J Lepr Other Mycobact Dis, 1990 Sep, 58(3), 480 - 90
Operational value of serological measurements in multibacillary leprosy patients: clinical and bacteriological correlates of antibody responses; Roche PW et al.; The antibody responses of 100 previously untreated multibacillary (MB) leprosy patients to one protein and two carbohydrate antigens were examined: 94% of the patients had Mycobacterium leprae-specific antibodies; 89% directed to the species-specific epitope on phenolic glycolipid (PGL-I), 89% against the specific epitope on the 35-kDa protein, and 94% against one or both of the two . By contrast, 67% of the patients had anti-lipoarabinomannan (LAM) antibodies . There were trends for the seropositivity rate and the antibody level to rise with the increasing extent of the disease and as patients moved to the polar lepromatous end of the spectrum . The bacillary load, as measured by the bacterial index, was moderately correlated with the IgM anti-PGL-I and the anti-35-kDa antibody levels and, to a lesser extent, with the IgG antibodies directed at the common mycobacterial carbohydrate LAM . The sensitivity of the IgM anti-PGL-I antibodies for detecting smear-positive MB disease was 91%; that for the anti-35-kDa antibodies was 92%.

J Urol, 1990 Sep, 144(3), 658 - 61
A randomized controlled study of intravesical alpha-2b-interferon in carcinoma in situ of the bladder; Glashan RW; We treated 87 patients with carcinoma in situ of the bladder in a prospective randomized trial of 2 dose levels of intravesically administered alpha-2b-interferon . Patients received either low dose (10 million units) or high dose (100 million units) recombinant alpha-2b-interferon weekly for 12 weeks and then monthly for a maximum of 1 year . Of the 47 high and 38 low dose patients 20 (43%) and 2 (5%), respectively, achieved a complete response . Additionally, partial responses (cytology results positive with no histological evidence of carcinoma in situ) were noted in 23% of the high dose group . Notably, 6 of 9 patients who had failed prior intravesical bacillus Calmette-Guerin therapy responded to alpha-2b-interferon treatment . Preliminary assessment has shown that among the complete responders 18 of 20 (90%) in the high dose group have maintained responses for at least 6 months after the completion of treatment (10 for more than 12 months) . Seven patients in each treatment group have undergone radical cystectomy . All 14 patients had progressive disease except 1 who chose cystectomy although she was still responding to treatment . The median intervals from initial treatment to cystectomy were 18 and 32 weeks in the low and high dose groups, respectively . Local irritation or toxicity did not occur and other adverse effects were rare except for mild to moderate flu-like symptoms (8% in the low dose and 17% in the high dose groups) . No patient discontinued therapy due to treatment-related adverse effects . Intravesical alpha-2b-interferon demonstrated a high level of activity in the treatment of carcinoma in situ of the bladder with the 100 million unit dose producing a significantly greater response rate (43% complete response, p less than 0.0001) than the low dose (5% complete response) . Safety and tolerance were excellent with no local irritative toxicity.

J Invertebr Pathol, 1990 Sep, 56(2), 237 - 42
An in vitro system for testing Bacillus thuringiensis toxins: the lawn assay; Gringorten JL et al.; A cell assay system was developed that allows Bacillus thuringiensis delta-endotoxins activated at high pH (10.5) to be tested in vitro without causing alkaline injury to target cells . The assay is carried out on a lawn of gel-suspended cells, requires only 1 microliter of sample per dose, and is quantitative, rapid, and sensitive . The threshold dose for toxicity of B . thuringiensis subsp . kurstaki HD-73 with IPRI-CF-1 cells was 24 pg protein . The assay is also very useful for identifying antibodies which inhibit toxicity and for detecting beta-exotoxin.

Gene, 1990 Sep 1, 93(1), 49 - 54
Hyperexpression of a Bacillus thuringiensis delta-endotoxin-encoding gene in Escherichia coli: properties of the product; Ge AZ et al.; Conditions for hyperexpression, in Escherichia coli, of the Bacillus thuringiensis var, kurstaki gene, cryIA9(c)73, encoding an insecticidal crystal protein, CryIA(c)73, were investigated by varying the promoter type, host cell, plasmid copy number, the second codon and number of terminators . The cryIA(c)73 gene was cloned into three E . coli expression vectors, pKK223-3 (Ptac promoter), pET-3a (P phi 10 promoter), and pUC19 (Ptac promoter) . The level of cryIA(c)73 expression was measured by ELISA and compared to total cellular protein over growth periods of 24 and 48 h . Maximum expression levels of 284 microgram CryIA(C)73/ml (48% of cellular protein) were obtained in shake flasks with the Ptac promoter in E . coli JM103 . Optimal conditions were found to be low-copy-number plasmid (pBR322 ori), 48 h of growth, in lon+ cells . A change of the gene's second codon to AAA can improve expression by two to three fold but is undetectable in the presence of a strong E . coli promoter . The cryIA(c)73 gene product, in E . coli, formed crystals with the same lattice structure as the native crystals formed in B . thuringiensis (as visualized by electron microscopy) . Bioassay results (insect toxicity and specificity) of the crystal produced in E . coli were similar to that produced in B . thuringiensis.

Cesk Epidemiol Mikrobiol Imunol, 1990 Sep, 39(5), 272 - 9
{Products of Bacillus cereus contributing to its pathogenicity}; Hostacka A; In a review the author describes the most important exoproducts of strains of Bacillus cereus which contribute to its pathogenicity . The toxic product are evaluated with regard to their physical, chemical and biological properties as well with regard to mechanism of their pathogenic action.

Mikrobiologiia, 1990 Sep-Oct, 59(5), 782 - 9
{Biosynthesis of fibrinolytic enzymes with different mechanisms of action by microorganisms of the genus Bacillus}; Vyrbornykh CN et al.; The effect exerted by the composition of a growth medium on the biosynthesis of fibrinolytic enzymes was studied in Bacillus licheniformis 125, B . macerans P1 and Bacillus sp . Fibrinolytic enzymes with the plasmin and activator effects on fibrin were found to be formed predominantly depending on the specified conditions, e.g . in a chemically defined medium with potato broth.

Ukr Biokhim Zh, 1990 Sep-Oct, 62(5), 103 - 6
{The effect of respiratory toxins on the bacterial concentration of trivalent gold}; Karamushka VI et al.; It has been found that concentration of trivalent gold by Bacillus cereus V-4368 cells is sensitive to respiratory inhibitors such as rotenone and antimycin A . Inhibitory action of rotenone is neutralized by succinate, that of antimycin--by ATP . ATP influence disappears in presence of DCCD . It is supposed that Au(III) concentration is due to the ATPase action and the respiration systems supply this process with ATP.

Nippon Saikingaku Zasshi, 1990 Sep, 45(5), 841 - 3
{Spore-spheroplast transformation of Bacillus megaterium}; Takubo Y et al.; A new method for transformation of Bacillus megaterium was developed by modification of Chang and Cohen's method . In our method, spore spheroplasts were used as recipient cells instead of the protoplasts of vegetative cells . Longer incubation (60 min) of spore spheroplasts and plasmid DNA before treatment with polyethylene glycol remarkably increased the efficiency of transformation . The frequency of transformation was about 10(4) per microgram of plasmid DNA . A shot-gun-type cloning of chromosome DNA of B . megaterium ATCC 12872 was available in B . megaterium ATCC 19213 strain by this transformation method.

J Appl Bacteriol, 1990 Sep, 69(3), 414 - 20
Effects of heat-, CaCl2- and ethanol-treatments on activation of Bacillus spores; Kim J et al.; The effects of heat, CaCl2, and ethanol on activation of Bacillus spores were determined by monitoring the absorbance decrease during germination in inosine . Bacillus cereus T, B . subtilis A and B . megaterium QM B1551 spores were activated by heat- and CaCl2-treatments . Ethanol activated B . megaterium and B . subtilis spores yet did not activate B . cereus spores . CaCl2- and ethanol-activations were less effective than heat-activation as judged by optimal germination rates and germination extents . The presence of CaCl2 during heat-treatment inhibited heat-activation of all three Bacillus spores without affecting viability or dipicolinic acid content of the spores . The electrophoretic patterns of coat plus outer membrane proteins extracted from Bacillus spores treated with CaCl2 and heat in the presence of CaCl2 were similar to each other and were distinctively different from the patterns of proteins from unactivated spores or the spores treated with heat and/or ethanol.

EMBO J, 1990 Sep, 9(9), 2743 - 50
The multifunctional peptide synthetase performing the first step of penicillin biosynthesis in Penicillium chrysogenum is a 421,073 dalton protein similar to Bacillus brevis peptide antibiotic synthetases; Smith DJ et al.; The nucleotide sequence of the Penicillium chrysogenum Oli13 acvA gene encoding delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase, which performs the first step in penicillin biosynthesis, has been determined . The acvA gene contains an open reading frame of 11,238 bp encoding a protein of 3746 amino acids with a predicted mol . wt of 421,073 dalton . Three domains within the protein of approximately 570 amino acids have between 38% and 43% identity with each other and share similarity with two antibiotic peptide synthetases from Bacillus brevis as well as two other enzymes capable of performing ATP-pyrophosphate exchange reactions . The acvA gene is located close to the pcbC gene encoding isopenicillin N synthetase, the enzyme for the second step of beta-lactam biosynthesis, and is transcribed in the opposite orientation to it . The intergenic region of 1107 bp from which the acvA and pcbC genes are divergently transcribed has also been sequenced.

Am Rev Respir Dis, 1990 Sep, 142(3), 508 - 11
Antituberculous drug resistance in central Haiti; Scalcini M et al.; To determine the prevalence of antituberculous drug resistance in Haiti, we conducted a 1-yr survey in a central district . From a bacillary positive (smear and/or culture positive) case rate of 80/100,000, there were 282 patients from whom Mycobacterium tuberculosis was cultured . Each isolate was packaged and delivered to Canada where speciation and drug susceptibility testing were performed . Reported resistances are those using the proportions method (Laboratory Center for Disease Control, Ottawa, Canada) . Resistance to one or more drugs was found in 22% of isolates . Age was the most important predictor of resistance in Haiti; resistance rates for age groups less than 14, 14 to 29, 30 to 44, greater than or equal to 45 were 8, 19, 22, and 31%, respectively . In patients not known to have received antituberculous drugs in the past, resistances were isoniazid (19%), streptomycin (5%), ethambutol (2%), ethionamide (2%), rifampin (1%) . We conclude that antituberculous drug resistance is prevalent in Haiti, especially in older age groups, and that in persons with no known antituberculous drug use in the past, resistance to isoniazid is significant.

Res Microbiol, 1990 Sep-Oct, 141(7-8), 931 - 9
Recombinant BCG as a candidate oral vaccine vector; Barletta RG et al.; Bacille Calmette-Guerin (BCG), currently the most widely used vaccine in the world, was originally administered for many years as an oral vaccine . The low frequency of serious complications, inexpensive production, and adjuvanticity make BCG an ideal candidate for a recombinant vaccine vehicle . Although mycobacteria are slow growing and not yet well characterized genetically, we have recently developed technology for the genetic manipulation of BCG and other mycobacteria . Phage and plasmid systems based on a shuttle strategy to manipulate DNA in Escherichia coli and transfer it to mycobacteria have been developed . We have established that the aminoglycoside phosphotransferase gene can be used as an effective selectable marker in the mycobacteria and that a foreign antigen from Mycobacterium leprae can be expressed in BCG . Furthermore, a thorough analysis of mycobacterial expression sequences has been undertaken to optimize the expression of foreign antigens in BCG . We constructed an expression probe shuttle plasmid with beta-galactosidase as reporter gene, and have used it successfully to identify multiple mycobacteriophage DNA sequences with varying levels of constitutive or regulable promoter activity . Further genetic advances required for development of recombinant BCG into an effective recombinant vaccine vehicle, including possibilities for oral administration, are adumbrated.

Hua Xi Yi Ke Da Xue Xue Bao, 1990 Sep, 21(3), 330 - 3
{Diagnostic value of transbronchial lung biopsy in diffuse or peripheral lung lesions}; Chen W et al.; The results of 102 cases of diffuse or peripheral lung lesions examined by transbronchial lung biopsy (TBLB), bronchial brushing (BB), and bronchial alveolar lavage (BAL) via fiberoptic bronchoscope, were reported . The positive diagnostic rate was 74.5% (76 cases) . In lung cancer, the positive rate by means of BB was 77.1%, which was higher than that by TBLB (58.1%) . In pulmonary tuberculosis, the positive rate by TBLB was 76.9%, higher than that by BAL fluid for identification of tubercle bacillus by culture (44.4%) . If TBLB was combined with BB and BAL, the positive diagnostic rate would be further elevated . The data showed that if the size of the masses greater than or equal to 3cm in peripheral lung field on chest film, the possibility of lung cancer was greater than that of those less than 3cm . Most cases of localized infiltration in the lungs were caused by tuberculosis . But the diffuse lesions of the lungs were often caused by bronchiolo-alveolar carcinoma, adenocarcinoma, diffuse interstitial fibrosis of the lungs, silicosis, sarcoidosis, etc . By the careful study of the chest film and ascertainment of the exact locations of the pulmonary lesions there, we can carry out the TBLB and obtain a satisfactory specimen without any X-ray monitoring . In the present group of patients who underwent TBLB, one was complicated by haemorrhage (greater than 50ml) and two by pneumothorax, but all of them recovered promptly after proper management . By strick adherence to indication, adequate preoperative preparations and very careful performance of the procedure, the complications of TBLB could be reduced to minimum.(ABSTRACT TRUNCATED AT 250 WORDS)

Southeast Asian J Trop Med Public Health, 1990 Sep, 21(3), 430 - 6
The effect of Bacillus thuringiensis israelensis {H-14} on emergence of Mansonia mosquitos from natural breeding habitat; Chang MS et al.; The measurement of the ultimate effects of the microbial insecticides on mosquito density is best obtained by assessment of adult populations . The main aims of this study are: (1) to assess the effect of Bacillus thuringiensis israelensis (Bti) FC Skeetal and Bactimos briquettes on the emergence rate of Mansonia bonneae developed from the introduced first-instar stage larvae and (2) to measure the effect of these two formulations of insecticides on Mansonia adult populations emerging from the natural breeding plots . Bti Skeetal and Bactimos briquettes at the lower applied dosages of 2.3 kg/ha and 1 briquette case/20 m2 respectively achieved 39-40% pupation rates and 31.5-34.2% adult emergence rates . At these low applied dosages, there was little or no direct effect on pupation from the surviving larvae and thereafter on the emergence of adults from the pupae . A two-fold increase in dosage, however, produced a drastic decline in the pupation rate and adult emergence rate . The rates dropped to 6.5% (pupation) and 4.3% (adult emergence) of the total larvae for Bactimos briquettes and to merely 1.5% (pupation) and 1.3% (adult emergence) of the total larvae for Skeetal . In studying the effect of Bti on the field populations of Mansonia mosquitos, two plots each were treated with Bactimos at 1 briquette case/10 m2 and Skeetal at 4.6 kg/ha . A wooden pyramid-shaped screened cage was placed on a cluster of host plants for a period of 2 weeks to trap the emerging adult mosquitoes . There were a total of 24 clusters of host plants in each plot.(ABSTRACT TRUNCATED AT 250 WORDS)

Protein Expr Purif, 1990 Sep, 1(1), 13 - 8
Purification of the temperature-specific surface antigen of Paramecium primaurelia with its glycosyl-phosphatidylinositol membrane anchor; Azzouz N et al.; The membrane form of the temperature-specific G surface antigen of Paramecium primaurelia strain 156 has been purified by a novel procedure utilizing solubilization by detergent, ammonium sulfate precipitation, and high-performance liquid chromatography . The surface antigen, which was prepared in a nondenatured state containing a glycosyl-phosphatidylinositol membrane anchor, migrated as a single band upon electrophoresis in sodium dodecyl sulfate-polyacrylamide gels . Following cleavage of the purified surface antigen by a phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis, the soluble form was released with the unmasking of a particular glycosidic immunodeterminant called the cross-reacting determinant . The purification protocol described here will now permit further biochemical and biophysical characterization of the nondenatured membrane form of Paramecium surface antigens.

Mikrobiol Zh, 1990 Sep-Oct, 52(5), 8 - 11
{The site-specific endonucleases of bacteria in the genus Bacillus}; Sorokulova IB et al.; Production regularities of site-specific endonucleases by aerobic spore-forming bacteria, separated from different ecological sources have been studied . It is shown that more than 1/3 of all the studied cultures produce site-specific endonucleases . A dependence of occurrence frequency of bacteria producers on the econiche of their separation has been noticed . The data on production of species-specific restrictases are obtained which can serve additional characteristics for differentiation of close species of Bacillus.

Biochem J, 1990 Sep 1, 270(2), 363 - 7
The poly-alpha- and -beta-1,4-glucuronic acid moiety of teichuronopeptide from the cell wall of the alkalophilic Bacillus strain C-125; Aono R; Teichuronopeptide is a structural component of the cell wall of alkalophilic Bacillus strain C-125 and is a complex composed of polyglutamate and polyglucuronate . A structural analysis of the polyglucuronic acid moiety was carried out . Periodate oxidation and Smith degradation of the moiety, and enzymic analysis after reduction of glucuronic acid to glucose, revealed that glucuronic acid bound together with alternately alpha- and beta-1,4-linkages.

Am J Clin Pathol, 1990 Sep, 94(3), 307 - 12
Polyclonal antibodies raised against Bacillus Calmette-Guerin, Mycobacterium duvalii, and Mycobacterium paratuberculosis used to detect mycobacteria in tissue with the use of immunohistochemical techniques; Wiley EL et al.; Commercially available polyclonal antibodies raised against strains of mycobacteria were used to detect organisms in tissue sections from 34 cases of tuberculosis, leprosy, and atypical mycobacteria . Thirty-two cases of fungal infections, granulomatous inflammation, and sarcoidosis were used as negative controls . Sections stained with the use of antibodies raised against Bacillus Calmette-Guerin (BCG), Mycobacterium duvalii (MD), and Mycobacterium paratuberculosis (MP) were compared with Kinyoun and Fite-stained tissue sections . In caseating granulomata, clumps of mycobacterial debris, cells, and cell fragments stained . In histiocytic granulomata of mycobacterial infections, histiocyte cytoplasm contained both organisms and debris . The three antibodies showed cross-reactivity against the four groups of mycobacteria tested . Mycobacterial staining using immunoperoxidase was apparent in most cases at low-power (scanning) magnification . Thirty-two of 34 cases of mycobacterial infection, including all 24 Kinyoun-Fite-positive cases, were positive for immunoreactive organisms and debris using anti-MD, anti-BCG, and/or anti-MP . Eight of ten cases of culture-proven mycobacterial infection, in which Kinyoun and Fite stains were negative, had immunoreactive organisms or antigen with anti-BCG, MD, or MP . The antibodies also stained organisms in five cases of sporotrichosis in which the organisms were identified as yeast forms in tissue sections.

J Biol Chem, 1990 Aug 25, 265(24), 14065 - 8
The major native proteins of the leprosy bacillus; Hunter SW et al.; This study addresses a major obstacle to vaccine development for leprosy, the isolation and characterization of the native protein antigens of the leprosy bacillus . Mycobacterium leprae harvested from armadillos was subjected to a simple fractionation protocol to arrive at the three major subcellular fractions, cell walls, cytoplasmic membrane, and soluble cytoplasm . The application of extensive detergent phase separations to membrane fractions allowed removal of lipoarabinomannan and the mannosyl phosphatidylinositols, and the recognition and purification of two major membrane proteins (MMP) of molecular mass 35 kDa (MMP-I) and 22 kDa (MMP-II); recovery of these proteins was about 0.5 mg each per g of M . leprae . MMP-I is N-blocked and is perhaps a lipoprotein . End group analysis on MMP-II indicates a new protein . Three major cytoplasmic proteins (MCP) of molecular mass 14 kDa (MCP-I), 17 kDa (MCP-II), and 28 kDa (MCP-III) were also recognized . MCP-I, the most abundant protein in M . leprae, represents 1% of the bacterial mass . End group analysis of the first 30 residues and immunoblotting studies demonstrate sizeable structural homology to a protein from Mycobacterium tuberculosis but immunological distinctiveness . MCP-I, which also occurs in highly immunogenic peptidoglycan-bound form, is a primary candidate for future vaccine development . The cell walls of M . leprae are also characterized by one major extractable protein, also of molecular mass 17 kDa . Thus the major antigens of the leprosy bacillus, protein and carbohydrate alike, are now nearer to complete definition.

FEBS Lett, 1990 Aug 20, 269(1), 69 - 72
Submicromolar Ag+ increases passive Na+ permeability and inhibits the respiration-supported formation of Na+ gradient in Bacillus FTU vesicles; Semeykina AL et al.; The effect of Ag+ on Na+ pumping by Na(+)-motive NADH-quinone reductase and terminal oxidase has been studied in Bacillus FTU inside-out vesicles . Very low concentrations of Ag+ (C1/2 = 1 x 10(-8) M or 2 x 10(-12) g ion.mg protein-1) are shown to inhibit the uphill Na+ uptake coupled to the oxidation of NADH by fumarate or of ascorbate + TMPD by oxygen but exert no effect on the H+ uptake by the H(+)-motive respiratory chain . Low Ag+ also induces a specific increase in the Na+ permeability of the vesicles . HQNO, added before and not after Ag+, prevents the Ag(+)-induced permeability increase, with effective HQNO concentrations being similar to those inhibiting the uphill Na(+)-uptake coupled to the NADH-fumarate oxidoreduction . Reduction of terminal oxidase by ascorbate + TMPD in the presence of cyanide sensitizes the Na+ permeability to Ag+ . It is suggested that low {Ag+}, known as a specific inhibitor of electron transport by the Na(+)-motive NADH-quinone reductase, uncouples the electron and Na+ transports so that the Ag(+)-modified NADH-quinone reductase operates as an Na+ channel rather than an Na+ pump . This effect is discussed in connection with the antibacterial action of Ag+.

J Mol Biol, 1990 Aug 20, 214(4), 807 - 9
Maltodextrin-dependent crystallization of cyclomaltodextrin glucanotransferase from Bacillus circulans; Lawson CL et al.; Crystals of cyclomaltodextrin glucanotransferase from Bacillus circulans (EC 2.4.1.19) suitable for high-resolution X-ray analysis were obtained by vapor diffusion against 60% (v/v) 2-methyl 2,4-pentanediol buffered with 100 mM-sodium Hepes, pH 7.55 . The crystals have P2(1)2(1)2(1) space group symmetry, with a = 120.4 A, b = 110.9 A and c = 66.4 A, and contain one molecule of 68,000 in the asymmetric unit . Growth of single enzyme crystals was found to require the presence of either alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, or maltose in high molar excess, a requirement that could not be fulfilled by glucose, the basic building block of these compounds . Although the exact role of cyclic and linear maltodextrins in enzyme crystallization is not yet known, we have preliminary evidence that these compounds are degraded by the enzyme in the crystallization droplet.

Biochem J, 1990 Aug 15, 270(1), 133 - 6
The construction of Bacillus thuringiensis strains expressing novel entomocidal delta-endotoxin combinations; Crickmore N et al.; Using our recently reported method of electroporation to transform Bacillus thuringiensis {Bone & Ellar (1989) FEMS Microbiol . Lett . 58, 171-178}, cloned B . thuringiensis entomocidal delta-endotoxin genes have been introduced into several native B . thuringiensis strains . In many cases the resulting transformants expressed both their native toxins and the cloned toxin, producing strains with broader toxicity spectra . The introduction of the var . tenebrionis toxin gene into B . thuringiensis var . israelensis resulted in a strain with activity against Pieris brassicae (cabbage white butterfly), an activity which neither parent strain possesses . We discuss further the possibility of synergism and also the problems associated with introducing cloned DNA by this method.

N Engl J Med, 1990 Aug 9, 323(6), 357 - 65
An investigation of the cause of the eosinophilia-myalgia syndrome associated with tryptophan use; Belongia EA et al.; BACKGROUND . The eosinophilia-myalgia syndrome is a newly recognized illness that has been associated with the consumption of tryptophan products . It is not known whether the cause is related to the tryptophan itself or to chemical constituents introduced by the manufacturing process . METHODS . To describe the epidemiology of the eosinophilia-myalgia syndrome further and elucidate a possible association with the manufacturing process, we conducted surveillance for the syndrome in Minnesota, a community survey of tryptophan use in Minneapolis-St . Paul, and a case-control study to assess potential risk factors, including the use of tryptophan from different manufacturers . We performed high-performance liquid chromatography on tryptophan samples to identify other chemical constituents . RESULTS . The prevalence of tryptophan use increased from 1980 to 1989 and was highest among women . Among the subjects for whom the source of the tryptophan was known, 29 of 30 case patients (97 percent) and 21 of 35 controls (60 percent) had consumed tryptophan manufactured by a single company (odds ratio, 19.3; 95 percent confidence interval, 2.5 to 844.9; P less than 0.001) . This company used a fermentation process involving Bacillus amyloliquefaciens to manufacture tryptophan . Analysis of the manufacturing conditions according to the retail lot demonstrated an association between lots used by case patients and the use of reduced quantities of powdered carbon in a purification step (odds ratio, 9.0; 95 percent confidence interval, 1.1 to 84.6; P = 0.014), as well as the use of a new strain of B . amyloliquefaciens (Strain V) (odds ratio, 6.0; 95 percent confidence interval, 0.8 to 51.8; P = 0.04) . There was a significant correlation (r = 0.78, P less than 0.001) between the reduced amount of powdered carbon used during manufacturing and the use of the new bacterial strain . High-performance liquid chromatography of this company's tryptophan demonstrated one absorbance peak (peak E) that was present in 9 of the 12 retail lots (75 percent) used by patients and 3 of 11 lots (27 percent) used by controls (odds ratio, 8.0; 95 percent confidence interval, 0.9 to 76.6; P = 0.022) . CONCLUSIONS . The outbreak of the eosinophilia-myalgia syndrome in 1989 resulted from the ingestion of a chemical constituent that was associated with specific tryptophan-manufacturing conditions at one company . The chemical constituent represented by peak E may contribute to the pathogenesis of the eosinophilia-myalgia syndrome, or it may be a surrogate for another chemical that induces the syndrome.

Appl Environ Microbiol, 1990 Aug, 56(8), 2480 - 3
Toxicity to Spodoptera exigua and Trichoplusia ni of individual P1 protoxins and sporulated cultures of Bacillus thuringiensis subsp . kurstaki HD-1 and NRD-12; Moar WJ et al.; The toxicities to neonate Spodoptera exigua and Trichoplusia ni of lyophilized powders obtained from sporulated liquid cultures (referred to as sporulated cultures) and Escherichia coli-expressed P1 {cryIA(a) cryIA(b) cryIA(c)} protoxins from three-gene strains of NRD-12 and HD-1 of Bacillus thuringiensis subsp . kurstaki were determined by using diet incorporation bioassays . Although sporulated cultures from both strains were more toxic to T . ni than S . exigua, there were no differences in toxicity between NRD-12 and HD-1 . Toxicities of the three individual P1 protoxins against S . exigua varied by at least fivefold, with the cryIA(b) protein being the most toxic . These same protoxins varied in toxicity against T . ni by at least 16-fold, with the cryIA(c) protein being the most toxic . However, when tested against either S . exigua or T . ni, there were no differences in toxicity between an NRD-12 P1 protoxin and the corresponding HD-1 P1 protoxin . Comparing the toxicities of individual protoxins with that of sporulated cultures demonstrates that no individual protoxin was as toxic to S . exigua as the sporulated cultures . However, this same comparison against T . ni shows that both the cryIA(b) and cryIA(c) proteins are at least as toxic as the sporulated cultures . Results from this study suggest that NRD-12 is not more toxic to S . exigua than HD-1, that different protein types have variable host activity, and that other B . thuringiensis components are not required for T . ni toxicity but that other components such as spores might be required for S . exigua toxicity.

J Gen Virol, 1990 Aug, 71 ( Pt 8), 1661 - 8
Comparative study of virion structure, protein composition and genomic DNA of three ascovirus isolates; Federici BA et al.; The virions of three ascoviruses isolated from the noctuids Heliothis zea, Spodoptera frugiperda and Trichoplusia ni were compared with respect to their size and structure, protein composition and the size and relatedness of their DNAs . The virions of the isolates from H . zea (HAV) and T . ni (TAV) were allantoid in shape (400 x 130 nm), enveloped and contained an inner particle which appeared to have an internal lipid bilayer surrounding the DNA core . The virions of the S . frugiperda isolate (SAV) were similar in structure and size, but were bacilliform in shape, and after formation, were often occluded in vesiculate occlusion bodies . In preparations of purified virions of each isolate, at least 12 polypeptides were detected that ranged in size from 10K to 200K and contained a major species of about 50K . The genome of SAV was about 140 kbp in size, whereas those of TAV and HAV were approximately 180 kbp . Analysis of DNA fragment patterns of the three isolates generated with BamHI, HindIII or XhoI, as well as DNA-DNA dot blot and Southern blot hybridization studies, demonstrated that HAV and TAV were closely related but not identical . The DNA from SAV, however, did not hybridize with the DNA from either of the other isolates . Thus the ascovirus isolates from T . ni and H . zea are considered variants of the same virus, whereas the isolate from S . frugiperda is a separate member of the ascovirus group.

Am J Med, 1990 Aug, 89(2), 216 - 22
Visceral bacillary epithelioid angiomatosis: possible manifestations of disseminated cat scratch disease in the immunocompromised host: a report of two cases; Kemper CA et al.; Opportunistic infection with the causative agent of cat scratch disease may be responsible for an unusual vascular proliferative lesion, referred to as bacillary epithelioid angiomatosis, previously described only in human immunodeficiency virus (HIV)-infected patients . We present a case of an HIV-infected patient with bacillary epithelioid angiomatosis involving the liver and bone marrow causing progressive hepatic failure . We also report a case of a cardiac transplant recipient with hepatic and splenic bacillary epithelioid angiomatosis manifesting as a fever of unknown origin, a previously unreported event in a non-HIV-infected patient . These cases represent the first documentation of bacillary epithelioid angiomatosis with visualization of cat scratch-like organisms involving internal organs.

Biochim Biophys Acta, 1990 Aug 1, 1040(1), 130 - 3
Studies on the interactions of glycerol dehydrogenase from Bacillus stearothermophilus with Zn2+ ions and NADH; Spencer P et al.; The interactions of the essential divalent cation, Zn2+, with the binary complex formed between glycerol dehydrogenase (glycerol:NAD+ 2-oxidoreductase, EC 1.1.1.6) and its coenzyme NADH have been examined by fluorescence spectroscopy . Both the metallo and non-metallo form of the enzyme bind the coenzyme NADH . The addition of Zn2+ ions to a solution of the binary complex formed between metal-depleted enzyme and NADH results in a rapid increase in fluorescence emission at 430 nm . This has been used to determine the on rate for Zn2+ to the enzyme/binary complex . A dissociation constant of 3.02 +/- 0.25.10(-9) M for the equilibrium between Zn2+ ions and the enzyme has been determined.

J Urol, 1990 Aug, 144(2 Pt 1), 313 - 5
The preparation, handling and use of intravesical bacillus Calmette-Guerin for the management of stage Ta, T1, carcinoma in situ and transitional cell cancer; Brosman SA et al.; The increasing use of bacillus Calmette-Guerin in the treatment of patients with low stage bladder cancer will inevitably bring attention to problems of proper use and toxicity . There is a need to identify patients who are most likely to benefit from this therapy and those who may be at risk for serious complications.

Acta Paediatr Jpn, 1990 Aug, 32(4), 357 - 60
Adverse effects on EEG and clinical condition after immunizing children with convulsive disorders; Nouno S et al.; 116 immunizations were given to 61 children with febrile convulsion or epilepsy who had not had a seizure for 1 year since the last attack . In 92 of the 116 immunizations the electroencephalogram (EEG) was examined before and after immunization . No adverse effects on the EEG were observed in 19 immunizations with Japanese encephalitis, measles, mumps or rubella vaccines . Epileptic spikes reappeared after 10 immunizations and epileptic spikes increased after 10 immunizations among 73 given for diphtheria, acellular pertussis and tetanus (DPT), diphtheria and tetanus (DT), or Bacillus Calmette-Guerin (BCG) . A convulsion was observed once in one child 7 days after immunization with BCG . A follow-up EEG examination is necessary after children with convulsive disorders are immunized.

Gig Sanit, 1990 Aug, (8), 18 - 22
{Survival of bacteria of the Bacillus and Pseudomonas species in phosphorus-polluted river and sea waters}; Alton LV; It has been stated, that phosphorus in the concentration 0.15 g/l, as an additional nutrition component in marine and river water produces a growth stimulating effect on certain types of bacteria . The concentration of 15 g/l, on the contrary, inhibits bacteria growth, and terms of their survival in marine and river water are reduced.

J Paediatr Child Health, 1990 Aug, 26(4), 212 - 6
Neonatal gram negative meningitis: a 10-year review, with reference to outcome and relapse of infection; Anderson SG et al.; Twenty-four infants treated for neonatal Gram negative bacillary meningitis over a 10-year period were reviewed to determine the mortality and incidence of complications including relapse . Nine (37.5%) infants died; two survivors had major and 13 minimal or no handicap . Five (21%) infants had a relapse of meningitis after the initial course of treatment; two of these infants had been treated with cefotaxime . The outcome did not correlate with age, sex, gestation, cerebrospinal fluid (CSF) parameters or peripheral neutrophil counts . A poor outcome was associated with thrombocytopenia, persistence of viable organisms in the CSF for more than 24 h and with seizures, particularly in infants aged 7 days or less . There were no clinical or laboratory parameters predictive of subsequent relapse and there was no apparent relationship with choice or dose of antibiotic used or duration of treatment.

Wei Sheng Wu Xue Bao, 1990 Aug, 30(4), 254 - 8
{Separation and purification of the toxic protein of Bacillus sphaericus Ts-1}; Yu Z et al.; Bacillus sphaericus strain Ts-1 is highly insecticidal to larvae of the mosquito . It's insecticidal component is toxic proteins . The toxin was extracted from spore-crystal complexes by disruption in a Sonicator Cell Disruptor Model W-220F followed by treatment with 0.05 mol/L NaOH . Fraction recovered from chromatography of the spore-crystal complexes on column of Sephadex G-200 were assayed against mosquito larvae and the toxic fractions from gel chromatography were subjected to SDS-PAGE . The toxic proteins in B . sphaericus Ts-1 spore-crystal complex migrated in position corresponding to 42kD and 43kD . Bioassay of the two purified proteins prepared by PAGE indicated that they were all toxic to mosquito larvae . Toxic protein was further purified by DEAE-cellulose chromatography . The toxic protein with a molecular weight of 42kD was obtained.

J Bacteriol, 1990 Aug, 172(8), 4247 - 54
Cloning, sequencing, and expression of a xylanase gene from the anaerobic ruminal bacterium Butyrivibrio fibrisolvens; Mannarelli BM et al.; A gene coding for xylanase activity, xynA, from the anaerobic ruminal bacterium Butyrivibrio fibrisolvens 49 was cloned into Escherichia coli JM83 by using plasmid pUC19 . The gene was located on a 2.3-kilobase (kb) DNA insert composed of two adjacent EcoRI fragments of 1.65 and 0.65 kb . Expression of xylanase activity required parts of both EcoRI segments . In E . coli, the cloned xylanase enzyme was not secreted and remained cell associated . The enzyme exhibited no arabinosidase, cellulase, alpha-glucosidase, or xylosidase activity . The isoelectric point of the cloned protein was approximately 9.8, and optimal xylanase activity was obtained at pH 5.4 . The nucleotide sequence of the 1,535-base-pair EcoRV-EcoRI segment from the B . fibrisolvens chromosome that included the xynA gene was determined . An open reading frame was found that encoded a 411-amino-acid-residue polypeptide of 46,664 daltons . A putative ribosome-binding site, promoter, and leader sequence were identified . Comparison of the XynA protein sequence with that of the XynA protein from alkalophilic Bacillus sp . strain C-125 revealed considerable homology, with 37% identical residues or conservative changes . The presence of the cloned xylanase gene in other strains of Butyrivibrio was examined by Southern hybridization . The cloned xylanase gene hybridized strongly to chromosomal sequences in only two of five closely related strains.

J Protein Chem, 1990 Aug, 9(4), 501 - 7
Subdomain organization of Bacillus thuringiensis entomocidal proteins' N-terminal domains; Chestukhina GG et al.; N-Terminal domain (65 kD) of delta-endotoxin produced by Bacillus thuringiensis ssp . alesti, as shown by limited proteolysis, consists of two subdomains of molecular mass 30 and 33 kD that correspond, respectively, to conservative and variable regions of the delta-endotoxin primary structure . Furthermore, proteolysis of these subdomains leads to their conversion into at least two fragments of molecular mass 10 kD stable to proteinase action . Such a pattern of molecular organization appears to be common for several structurally related delta-endotoxins that belong to the kurstaki group . Entomicidal protein produced by ssp . israelensis (70 kD), which differs strongly from alesti and other kurstaki group delta-endotoxins, retains a similar type of molecular organization and consists of two subdomains with molecular mass of approximately 35 kD . Apparently, the characteristic pattern of the delta-endotoxins' molecular structure reflects separation of functions (e.g., host recognition and toxicity per se) between domains and subdomains of these proteins.

J Appl Bacteriol, 1990 Aug, 69(2), 241 - 6
A simple haemolytic method for quantitation of the delta endotoxin of Bacillus thuringiensis subsp . israelensis from crude samples; Majumdar MK et al.; A simple haemolytic assay method for quantitative estimation of the delta endotoxin of Bacillus thuringiensis subsp . israelensis from a crude preparation has been developed . The method has several advantages over mosquito-larvicidal methods of assay as it is inexpensive, highly sensitive and easier to run and can be used for performing a reasonably large number of assays rapidly with high precision and with a coefficient of variation that does not exceed 1.96%.

J Biochem (Tokyo), 1990 Aug, 108(2), 235 - 40
Purification and some properties of a novel alpha-L-fucosidase capable of acting on alpha-(1----6)-L-fucosidic linkages from Bacillus circulans M28; Tsuji Y et al.; Two types of alpha-L-fucosidase (F-I and F-II), that differ in substrate specificity, were produced in the culture fluid by Bacillus circulans isolated from soil when the bacterium was cultivated on medium containing porcine gastric mucin . F-I was able to cleave the alpha-(1----2), alpha-(1----3), and alpha-(1----4)-L-fucosidic linkages in various oligosaccharides and glycoproteins, but not p-nitrophenyl alpha-L-fucoside, as previously reported {Y . Tsuji et al . (1990) J . Biochem . 107, 324-330} . F-II was purified from the culture fluid obtained with glucose medium by ammonium sulfate fractionation and various subsequent column chromatographies . The purified enzyme was found to be homogeneous on PAGE and its molecular weight was estimated to be approximately 250,000 . The maximal activity was observed between pH 6.0 to 7.0, the stable pH range being 6.0 to 8.5 . The enzyme specifically cleaved alpha-L-fucosidic bonds in low molecular weight substrates . The enzyme cleaved not only p-nitrophenyl alpha-L-fucoside, but also 2-fucosyllactose and 3-fucosyllactose . The enzyme was also able to act on the alpha-(1----6)-L-fucosidic linkages to N-acetylglucosamine in 6-O-alpha-L-fucopyranosyl-N-acetylglucosamine, and bi- and tetra-antennary oligosaccharides derived from porcine pancreatic lipase, which were not hydrolyzed by F-I.

Offentl Gesundheitswes, 1990 Aug-Sep, 52(8-9), 496 - 500
{Analysis of the epidemiological tuberculosis status in south Baden in the years 1986-1989--current tasks of the public health department in the campaign against tuberculosis}; Jancik M; All cases of tuberculosis reported in the districts Freiburg-City, Breisgau-Hochschwarzwald, Emmendingen, Lorrach and Ortenau in the years 1986/87 and 1988/89 have been analysed by a simple questionnaire . In both periods--86/87 and 88/89--an increasing number of cases of advanced pulmonary tuberculosis during the last two years, in particular among the younger population was detected . The bacillary form of pulmonary tuberculosis was notified in 55% of all reported cases in the years 86-89; before that, the highest rate was 44% . During the 2-two year period the number of registered cases of pulmonary tuberculosis did not change, patients were recorded 258 in the years 86/87 and 251 cases in 88/89 . The great majority of the patients was detected on the basis of subjective complaints . In the last two years, however, (88-89) there was a significant increase in converters in the environment of Tb cases and also--as compared with the years 86/87--more cases of pulmonary tuberculosis in children and adults among the examined contact persons . It is suggested to evolve a strategic plan for the elimination of tuberculosis similar to the procedure in the United States.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1990 Aug, 23(3), 211 - 9
Detection of mycobacterial antigens in sputum by enzyme-linked immunosorbent assay; Wu CH et al.; Enzyme-linked immunosorbent assay (ELISA) was evaluated for its usefulness in detecting mycobacterial antigens in sputum . Using anti-mycobacterial saline extract (MSE) and commonly available anti-bacillus Calmette Guerin (BCG) antibodies, Mycobacterium tuberculosis antigens can be detected in sputum by the double-antibody sandwich ELISA . A total of 1904 sputum specimens were tested by acid-fast bacillus (AFB) smear, culture and ELISA . Of 249 pulmonary tuberculosis patients, 225 (90.4%), 167 (67.1%), 114 (45.8%), 103 (41.4%) and (36.9%) were positive by BACTEC 460, 7H11 culture, AFB smear, BCG-ELISA and MSE-ELISA, respectively . If BACTEC 460 positive culture only is considered as the standard, sensitivity of 43.6% and 36.9%, and specificity of 98.2% and 98.3% are achieved, with positive predictive value of 76.0% and 74.1%, and negative predictive value of 92.8% and 92.1% for BCG- and MSE-ELISA, respectively . In 7H11 culture positive cases: 46.7% (BCG) and 43.1% (MSE) for sensitivity, 97.1% (BCG) and 97.7% (MSE) for specificity, 60.5% (BCG) and 64.3% (MSE) positive, 95% (BCG) and 94.7% (MSE) negative predictive values were achieved . When there is an urgent need the ELISA can serve as an alternative aid for the presumptive differentiation of other mycobacteria from M . tuberculosis.

Immunobiology, 1990 Aug, 181(1), 40 - 50
Staphylococcal enterotoxin A induced interferon (IFN)-gamma production in spleen cells from BCG-immunized mice: the IFN production is dependent on leukotriene C4 but not dependent on interleukin 2; Kato K et al.; In our previous paper, we showed that IFN was induced in sera by injection of staphylococcal enterotoxin A (SEA) in Bacillus Calmette-Guerin (BCG) immunized C57BL/6 (B6) mice . In analyzing the phenomenon in vitro, we showed that SEA induced IFN-gamma in the supernatant of the spleen cell culture from BCG immunized B6 mice and that leukotriene C4 (LTC4) from BCG activated macrophages in the spleen was involved in the IFN production from Ly 1+ T cells . On the other hand, interleukin-2 (IL-2) has reported to play an important role in the regulation of synthesis of IFN-gamma by T cells . In the present study, we examined whether IL-2 is involved in SEA-induced IFN production . The result showed that the SEA-induced IFN-gamma production was observed in spite of suppression of SEA-induced IL-2 production in spleen cells from BCG-immunized B6 mice . On the contrary, the depressed IFN production was observed in spite of high SEA-induced IL-2 production in spleen cells from their control mice . On the other hand, LTC4 production was 8 times higher in spleen cells from BCG-immunized B6 mice, high producer of SEA-induced IFN, than in that from BCG-immunized C3H mice, the low producer . We also observed that the IFN and the LTC4 production of spleen cells from BCG-immunized B6 mice was suppressed in the presence of caffeic acid and nordihydroguaiaretic acid, non-specific lipoxygenase inhibitors, and that LTC4 augmented the IFN production of normal B6 mouse spleen cells in the presence of 2-mercaptoethanol . Therefore, involvement of LTC4 rather than of IL-2 was supported in our experimental system.

Int J Food Microbiol, 1990 Aug, 11(1), 21 - 33
The effect of glucose, starch, and pH on growth, enterotoxin and haemolysin production by strains of Bacillus cereus associated with food poisoning and non-gastrointestinal infection; Garcia-Arribas ML et al.; Brain-Heart Infusion medium, modified by (a) 1.0% w/v glucose supplement, (b) 1.0% w/v soluble starch supplement, (c) pH adjustment to 8.8 or (d) pH adjustment to 5.0, was used to investigate the influence of glucose, starch and pH on growth, enterotoxin and haemolysin production by Bacillus cereus . The four test strains selected for comparison originated from episodes of emetic-syndrome food poisoning, diarrhoeal-syndrome food poisoning, traumatic wound infection and bovine mastitis, respectively . In the presence of either glucose or starch, growth of all strains was found to be near-optimum and accompanied by enhanced enterotoxin production . However, under the moderately acid and alkaline test conditions B . cereus strains exhibited varying degrees of growth inhibition and partial repression of enterotoxin and haemolysin production . In this respect the food-poisoning isolates proved to have greater resistance to adverse pH environments than did the non-gastrointestinal infection isolates.

Antimicrob Agents Chemother, 1990 Aug, 34(8), 1590 - 2
Cloning and sequencing of the class B beta-lactamase gene (ccrA) from Bacteroides fragilis TAL3636; Rasmussen BA et al.; Bacteroides fragilis TAL3636 produces a class B, Zn2(+)-requiring beta-lactamase . The gene, ccrA, was cloned and expressed in Escherichia coli . The gene was sequenced and shown to share greater than 33% identity with the metalloenzyme from Bacillus cereus 569/H.

Appl Environ Microbiol, 1990 Aug, 56(8), 2282 - 6
Plasmid-associated sensitivity of Bacillus thuringiensis to UV light; Benoit TG et al.; Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B . thuringiensis strains or of the closely related Bacillus cereus . Introduction of B . thuringiensis plasmids into B . cereus by cell mating increased the UV sensitivity of the cells and spores . Protoxins encoded by one or more B . thuringiensis plasmids were not involved in spore sensitivity, since a B . thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures . In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B . thuringiensis all increased the UV sensitivity of the spores . Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores . The cells of some strains harboring only B . thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores.

An Med Interna, 1990 Aug, 7(8), 406 - 10
{An analysis of the mortality from extrapulmonary tuberculosis in the catchment areas of the community of Valencia (1976-1980)}; Morales Suarez-Varela MM et al.; Tuberculosis is a chronic infectious disease produced by Mycobacterium tuberculosis or Koch's bacillus, which mainly affects the airways and, less frequently, other organs in the body . Tuberculosis is still a health and social problem in Spain . The data of this study were obtained from "Monografias Sanitarias", "Analisis de Mortalidad", published by "Generalitat Valenciana", the standard mortality rate was calculated by direct methods . Mortality is not the best sanitary parameter to study the real situation of this disease, because the present methods of treatment are effective and death caused by tuberculosis is very rare . Despite the aforementioned fact, when comparing the mortality data of different health areas during a period between 1976 and 1980, we confirm that this disease persists as a cause of death in our community.

Zhonghua Nei Ke Za Zhi, 1990 Aug, 29(8), 482 - 4, 511
{Effect of fluoroquinolones made in China in the treatment of acute bacillary dysentery . A report of 212 cases}; Wang XF et al.; 212 cases of acute bacillary dysentery were treated with enoxacin and norfloxacin, another 15 cases were treated by chujunsheng . The cure rate in enoxacin group was 99.1%, in norfloxacin group 98.04% and in chujunsheng group 63.64% . The isolated pathogens were tested for susceptibility with disk diffusion and MIC determination . The results showed that both enoxacin and norfloxacin are better than gentamycin, ampicillin, chloramphenicol and trimethoprim-sulfamethoxazole . Side effects of enoxacin group and norfloxacin were noted in are 2.7% and 3.0% of the patients respectively . This study suggests that enoxacin and norfloxacin are highly effective, quite convenient for use and have few adverse reactions . They are considered to be drugs of choice for the treatment of acute bacillary dysentery.

J Econ Entomol, 1990 Aug, 83(4), 1280 - 5
Efficacy and persistence of Bacillus sphaericus, Bacillus thuringiensis var . israelensis, and methoprene against Culiseta incidens (Diptera: Culicidae) in tires; Kramer VL; The efficacy and persistence of Bacillus sphaericus (2362) was compared at three dosage rates in tires that continually contained cadavers of Culiseta incidens (Thomson) versus tires with all dead larvae removed . At treatment rates of 3.75 and 7.5 ppm, the continual presence of cadavers in the tire water resulted in higher mortality rates . At the 15 ppm treatment rate, mortality rates were similar in tires with or without cadavers . Mortality rates increased in all tires 4-6 wk after treatment, suggesting an amplification of the pathogen . The mortality rate did not exceed 90% for greater than 2 wk in any of the tires . At a second site, the efficacy of B . sphaericus, Bacillus thuringiensis var . israelensis, and methoprene was evaluated against C . incidens in tires exposed to full sunlight versus shaded tires . In shaded tires inoculated with B . sphaericus (15 ppm) and B . thuringiensis var . israelensis (15 ppm), mortality exceeded 90% for 5 and 2 wk, and 50% for 10 and 4 wk, for the two bacteria, respectively . Larvae were adequately controlled (greater than 75% mortality) in the sunny tires for approximately 1 wk . The insect growth regulator, methoprene (applied at 2.5 ppm), inhibited the emergence of approximately 90% of the larvae present at the time of treatment, but not of larvae subsequently introduced into either the sunny or shaded tires.

Appl Microbiol Biotechnol, 1990 Aug, 33(5), 519 - 23
Characterization of an alkaline protease from Bacillus sp . no . AH-101; Takami H et al.; The Bacillus sp . no . AH-101 alkaline protease showed higher hydrolysing activity against insoluble fibrous natural proteins such as elastin and keratin in comparison with subtilisins and Proteinase K . The optimum pH of the enzyme toward elastin and keratin was pH 10.5 and pH 11.0-12.0 respectively . The specific activity toward elastin and keratin was 10,600 units/mg protein and 3970 units/mg protein, respectively . The enzymatic activity was not inhibited by p-chloromercuribenzoic acid and iodoacetic acid . Carbobenzoxy-glycyl-glycyl-L-phenylalanyl chloromethyl ketone completely inhibited the caseinolytic activity, but 36% elastolytic activity remained . No inhibitory effect on caseinolytic and elastolytic activity was shown by tosyl-L-phenylalanyl-chloromethyl ketone, tosyl-L-lysine chloromethyl ketone, carbobenzoxy-L-phenylalanyl chloromethyl ketone, and elastatinal . The amino acid composition and amino terminal sequence of the enzyme were determined . The no . AH-101 alkaline protease was compared with subtilisin BPN', subtilisin Carlsberg, no . 221, and Ya-B alkaline proteases . Extensive sequence homology existed among these enzymes.

Appl Microbiol Biotechnol, 1990 Aug, 33(5), 542 - 6
Molecular cloning, nucleotide sequence and expression in Escherichia coli of the beta-cyclodextrin glycosyltransferase gene from Bacillus circulans strain no . 8; Nitschke L et al.; The beta-cyclodextrin glycosyltransferase (beta-CGTase) gene was isolated from a lambda-library prepared from Bacillus circulans strain no . 8 . It was subcloned into plasmid pTZ and expressed by its endogenous regulatory sequences in Escherichia coli JM 103 . The structural gene was sequenced and showed an open reading frame for a polypeptide of 718 amino acid residues . The recombinant beta-CGTase had the same enzymatic properties as the extracellular CGTase (684 amino acid residues, corresponding to a mol . wt . of 74416) produced by B . circulans strain no . 8 . The amino acid sequence showed the highest homology (74.6% identical amino acids) with the CGTase of B . circulans strain F-2, which had been erroneously described as an amylase . The homology with the enzyme from the alkalophilic Bacillus sp . strain no . 1011 was 71.4% . The amino acid sequence derived will be used for elucidating the three-dimensional structure of the enzyme.

Biochemistry, 1990 Jul 31, 29(30), 7101 - 6
Probing the coenzyme specificity of glyceraldehyde-3-phosphate dehydrogenases by site-directed mutagenesis; Corbier C et al.; By combining our knowledge of the crystal structure of the glycolytic NAD-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the sequence of the photosynthetic NADP-dependent GAPDH of the chloroplast, two particular amino acid residues were predicted as the principal determinants of differing coenzyme specificity . By use of site-directed mutagenesis, the amino acids Leu 187 and Pro 188 of GAPDH from Bacillus stearothermophilus have been replaced with Ala 187 and Ser 188, which occur in the sequence from the chloroplast enzyme . The resulting mutant was shown to be catalytically active not only with its natural coenzyme NAD but also with NADP, thus confirming the initial hypothesis . This approach has not only enabled us to alter the coenzyme specificity by minimal amino acid changes but also revealed factors that control the relative affinity of the enzyme for NAD and NADP.

Biochem Biophys Res Commun, 1990 Jul 31, 170(2), 665 - 72
Purification and characterization of Bacillus thuringiensis var . tenebrionis insecticidal proteins produced in E . coli; MacIntosh SC et al.; Native and single amino acid variants of the Bacillus thuringiensis var . tenebrionis insecticidal proteins were expressed in Escherichia coli, purified and examined for biological and biochemical properties . A novel, pH dependent, preferential precipitation method was implemented to purify Escherichia coli produced Bacillus thuringiensis var . tenebrionis proteins, which are active against Colorado potato beetle (Leptinotarsa decemlineata) larvae . Cysteine residues of the native Bacillus thuringiensis var . tenebrionis protein were replaced by serine residues by site-directed mutagenesis to investigate the biological and structural importance of the individual cysteine residues . Sulfhydryl determination of the native and amino acid variant Bacillus thuringiensis var . tenebrionis proteins revealed that the native protein contains no disulfide bonds . Modification of the carboxyl terminal cysteine residue (amino acid 540) caused complete inactivation of the protein . Native, truncated and single amino acid variants (other than at amino acid 540) exhibited insecticidal activities comparable to each other and to solubilized crystals from the original strain.

Biochem J, 1990 Jul 15, 269(2), 507 - 12
Comparative analysis of the individual protoxin components in P1 crystals of Bacillus thuringiensis subsp . kurstaki isolates NRD-12 and HD-1; Masson L et al.; Two commercially important strains (NRD-12 and HD-1) of the entomopathogenic bacterium Bacillus thuringiensis subsp . kurstaki each contain three genes of partially identical sequence coding for three classes of 130-135 kDa protoxins (termed the 4.5, 5.3 and 6.6 protoxins) that display toxicity towards various lepidopteran larvae . These gene products combine to form the intracellular bipyramidal P1 crystal . Each of the genes from both strains was cloned and expressed in Escherichia coli . Analysis of the cloned genes at the restriction-endonuclease level revealed no detectable differences among genes within a particular gene class . The composition of the P1 crystal from both strains was quantitatively analysed by CNBr cleavage of the purified P1 crystal, with the purified recombinant gene products as reference proteins . Independent verification of the presence of high 6.6-protoxin gene product in the P1 crystal was provided by a rapid in vitro lawn cell toxicity assay directed against a Choristoneura fumiferana (CF-1) insect cell line . The results indicate that, although all three gene products are represented within the P1 crystal of either NRD-12 or HD-1, only the contents of the 4.5 and 5.3 protoxins vary between the two crystals, whereas the 6.6 protoxin contents are similar and represent approximately one-third of the P1 crystal in either strain.

Biochim Biophys Acta, 1990 Jul 6, 1039(3), 331 - 8
Purification and characterization of N-acetylmuramoyl-L-alanine amidase from human serum; Vanderwinkel E et al.; Purification to homogeneity of the N-acetylmuramoyl-L-alanine amidase (mucopeptide amidohydrolase, EC 3.5.1.28) from human serum has been achieved with a high yield . By molecular sieving chromatography, a molecular weight of 120,000-130,000 has been found for the native enzyme . Polyacrylamide gel electrophoresis under native conditions gave a unique band of Mr = 125,000 . The same technique performed under denaturing conditions revealed that the protein is a dimer composed of one subunit of Mr = 57,000 and another of Mr = 70,000 . In isoelectrofocalization assays, the amidase behaved as an acidic protein . Ethylenediaminetetraacetate inhibited the enzyme activity; the Mg2+ requirement was confirmed . The simultaneous presence of sulfhydryl groups and disulfide bonds in the protein was evidenced by the inhibitions produced by different thiol-blocking reagents and by several thiol-bearing substances . Direct measurements established the presence of two accessible thiol groups and the occurrence of nine disulfide bonds per protein molecule . Studies of substrate hydrolyzing capacities showed a marked preference for the muramoyl tripeptide derived from the Escherichia coli or Bacillus cereus mureins, the disaccharide tetrapeptide and the bis disaccharide tetra-tetrapeptide from E . coli were also good substrates . Activities on small muropeptides of other composition are also reported . Whole (insoluble) peptidoglycans representing the main bacterial chemotypes were submitted to the enzyme action; although with weak specific activities, the human amidase was nevertheless able to release soluble peptides from some of them . A bacteriolytic capacity on some microorganisms cannot be excluded . Results are discussed and the human enzyme is compared to presently known microbial muramoyl amidases.

Gaoxiong Yi Xue Ke Xue Za Zhi, 1990 Jul, 6(7), 344 - 9
{Applications of bacterial pathogens to the mosquito control}; Lu KH et al.; Both Bacillus thuringiensis subsp . isrealensis (Bti) and Bacillus sphaericus are common bacteria found in a variety of soil and aquatic habitats . Some strains of them can produce a protein crystal, called parasporal body (or crystal) beside the spore in the cell during sporulation, and it is toxic if eaten by mosquito larvae . As the parasporal bodies are ingested into a mosquito digestive tract, they will be digested and activated by certain enzymes in alkaline digestive juice . The activated toxins can destroy midgut epithelium and other tissues, and then the mosquitoes will be intoxicated and killed . There are some factors affecting efficiency of the toxins produced by these two mosquitocidal bacteria . First, the toxicity of the toxins will decrease as larval age and density increase . Second, difference in mosquito species will demonstrate a different tolerance to both toxins . However, larvae of Anopheles are more tolerant to Bti than those of Aedes and Culex; whereas larvae of Aedes are more tolerant to B . sphaericus than Culex and Anopheles . Environmental factors are as important as the mosquito hosts in influencing the efficiency of the toxins . For examples, raising temperature under suitable ranges may increase the toxicity of the toxins . The UV light of sunlight could inhibit activity of the toxins and therefore decrease their efficiency . Quality and depth of the water are also important environmental factors affecting toxicity of the toxins to the mosquito larvae . The precipitation of the crystal and organic materials absorbing the toxins decreases their toxicity to the mosquito larvae as well.

Gaoxiong Yi Xue Ke Xue Za Zhi, 1990 Jul, 6(7), 330 - 6
{Application and environmental impact of the biocontrol agent--Bacillus thuringiensis subsp . israelensis for vector control}; Hsu TH; In 1976, Goldberg and Margalit found a potent isolate of Bacillus thuringiensis subsp . israelensis, which has great mosquitocidal activity and was commercialized within 5 years in the USA . It is now the only bio-control agent on the market for vector control worldwide . Because of concerns about environmental pollution and ecological preservation, vector control can no longer be solely dependent on the use of chemicals . It appears that the best strategy is to be integrated with biological control . When considering the value and future progress of BTI, it is important to evaluate applicability based on (1) production technology and economic considerations (2) standardization and quality control, and (3) field realization and environmental impacts . Finally, government policy and regulation limitations are the key factors which influence the availability of BTI in the vector control program.

Jikken Dobutsu, 1990 Jul, 39(3), 425 - 8
Differences in susceptibility to peroral inoculation with Bacillus piliformis spores in rats and mice; Itoh T et al.; The median liver lesion producing doses of peroral inoculation with the spores of Tyzzer's organism RJ strain were 10(4 . 3) in rats and 10(2 . 7) in rats receiving prednisolone treatment for the provocation of Tyzzer's disease . In contrast to rats, liver lesions were detected in few mice inoculated perorally with 10(7) spores . In mice inoculated perorally with 10(7) spores, excretion of infective spores in the feces was detected only on day 1 postinoculation . On the other hand, no difference in susceptibility between rats and mice was detected upon intravenous inoculation with vegetative cells of the RJ strain . These results suggest that germination of the spores in the intestinal tract causes the difference in the susceptibility in rats and mice.

Jikken Dobutsu, 1990 Jul, 39(3), 371 - 6
{Heat and ozone resistance of Bacillus spores isolated from laboratory animals}; Pan TM et al.; Heat resistance of free-spores of 78 Bacillus strains isolated from laboratory animals was examined . Spores of 41 out of 78 strains survived for 320 minutes at 70 degrees C, 27 for 160 min, at 100 degrees C, only one for 20 min . at 110 degrees C by autoclaving, and none for 5 min . at 120 degrees C . D-values at 100 degrees C of 9 strains determined were between 5.03 and 30.06 min . Spores of 9 strains from stock cultures were exposed to ozone gas at various conditions . Ozone resistance of spores was closely dependent upon relative humidity . D-values of the spores tested by treatment with 200 ppm ozone at 60% RH were over 200 min., especially over 1,000 min . in 4 strains, indicating that exposure to ozone at a moderate humidity for 6 hours could not sterilize Bacillus spores . At 90% RH, however, treatment with 200 ppm ozone for 6 hr . might be effective for a routine sterilization in laboratory animal facilities.

Biomed Environ Mass Spectrom, 1990 Jul, 19(7), 395 - 9
Sequence determination of unknown cyclic peptide antibiotics by fast atom bombardment mass spectrometry; Ishikawa K et al.; The primary structures of unknown cyclic peptides produced by a Bacillus strain have been determined by fast atom bombardment (FAB) mass spectrometry, which has established the peptides as a new family of the iturin group antibiotics . FAB mass spectra of the intact peptides gave the immonium ions characteristic of constituent amino acids which made it possible to distinguish Asn from Asp and Glu from Gln . Collision-induced dissociation (CID) spectra of protonated molecules provided complete information on the connectivity of amino acid residues but did not reveal the direction of peptide bonding, while those obtained for fragment ions allowed us to make a discrimination between the correct sequence and its retro sequence . The amino acid sequences derived are c(Thr-Xaa-Asn-Tyr-Asn-Ser-Glu-Ser) (Xaa: C14 or C15 beta-amino acid) which are closely related to that of bacillomycin L . The structure is confirmed by the FAB mass spectra of the partial acid hydrolysate and the peptide mixture obtained from its single-step Edman degradation . Fragmentation processes involved in the CID spectra of the cyclic peptides are discussed based on the established amino acid sequence.

Int J Syst Bacteriol, 1990 Jul, 40(3), 242 - 6
Bacillus thiaminolyticus sp . nov., nom . rev; Nakamura LK; The name "Bacillus thiaminolyticus" Kuno 1951 was not included on the Approved Lists of Bacterial Names and has lost standing in bacteriological nomenclature . The genetic homogeneity of "Bacillus thiaminolyticus" was assessed by determining guanine-plus-cytosine contents by the buoyant density method and by measuring DNA relatedness by using spectrophotometric reassociation procedures . Of the 26 strains which I studied, 24 had guanine-plus-cytosine contents in the range from 52 to 54 mol% . The consistently high DNA relatedness values of 60 to 100% of these 24 strains to the type strain indicated that the "B . thiaminolyticus" group is genetically homogeneous . Low DNA relatedness values of 20 to 31% showed that "B . thiaminolyticus" is genetically unrelated to Bacillus alvei, "Bacillus aneurinolyticus," "Bacillus apiarius," Bacillus larvae, Bacillus laterosporus, Bacillus macerans, and Bacillus stearothermophilus . In general, the "B . thiaminolyticus" group was highly homogeneous for 49 phenotypic characteristics and clearly distinguishable from B . alvei, with which it was allegedly synonymous . On the basis of these findings, revival of the name Bacillus thiaminolyticus is proposed.

Br J Urol, 1990 Jul, 66(1), 35 - 9
Clinical study of prognostic factors of superficial bladder cancer treated with intravesical bacillus Calmette-Guerin; Shinka T et al.; Intravesical instillation of Tokyo 172 strain Bacillus Calmette-Guerin (BCG) was performed in 96 patients with initial superficial bladder cancer (Ta and T1) after transurethral resection (TUR) of tumour as a prophylaxis against tumour recurrence . The recurrence rate of tumours was estimated by the person-years method, comparing it with that of historical controls . There were statistically significant decreases in recurrent tumours following BCG therapy . To clarify the efficacy of intravesical BCG therapy, the prognostic significance of several factors was evaluated in patients with bladder cancer treated with TUR and instillation of BCG . The prophylactic effects were statistically better for those with multiple tumours, grade 3 lesions or Ta lesions than for control patients . No correlation between purified protein derivative (PPD) responsiveness and favourable results could be observed . Our results suggest that intravesical BCG instillation can alter the biological behaviour that affects the recurrence of superficial bladder cancer, especially for multiple, high grade or Ta tumours.

Immunology, 1990 Jul, 70(3), 344 - 50
Studies on the immunological properties of oxysterols: in vivo actions of 7,25-dihydroxycholesterol upon murine peritoneal cells; Moog C et al.; Oxygenated derivatives of cholesterol are potent immunosuppressors . It has been reported previously that 7,25-dihydroxycholesterol (7,25-OHC), synthesized in the URA31, strongly inhibits the early steps of T-cell activation . So far, the mechanisms underlying this type of effect have been mainly investigated in vitro, and the activity of these substances on the immune system has been poorly studied . This study describes that a single i.p . injection of 7,25-OHC induces a strong inflammatory response, consisting of a massive influx of macrophages and neutrophils into the abdominal cavity . Macrophages harvested from 7,25-OHC-treated mice express class II antigen to a lesser extent . Moreover, the 7,25-OHC treatment abolishes the class II induction by bacillus Calmette-Guerin (BCG) or interferon-gamma (IFN-gamma) . The inflammatory process triggered by the oxysterol is not the consequence of a non-specific effect due, for instance, to the presence of crystals in the abdominal cavity . Moreover, treatments by inhibitors of the acid arachidonic cascade do not affect the peritoneal exudate cell (PEC) influx induced by these substances . This study could be an important contribution to the mechanism determining the oxysterol-induced immunosuppression.

Am Rev Respir Dis, 1990 Jul, 142(1), 24 - 8
The influence of Calmette-Guérin bacillus immunization on the booster effect of tuberculin testing in healthy young adults; Sepulveda RL et al.; The booster or enhancement effect of repeated tuberculin skin testing in Calmette-Guerin bacillus (BCG)-vaccinated young adults was studied in 208 first-year medical, nursing, and medical technology students in Santiago, Chile, where BCG vaccine is usually administered at birth and at 6 and 14 yr of age . Thirty-three students had no BCG scar, 62 had one scar, 71 had two scars, and 42 had three scars . The mean age for each group was 19 yr . All students were healthy and had no known exposure to tuberculosis or history of tuberculosis or other mycobacterioses . The size in millimeters of induration of the first tuberculin reaction (PPD1) was clearly correlated with the number of BCG scars: 2.3 +/- 4.6 for no scars; 6.7 +/- 6.7 for one scar; 10.9 +/- 5.9 for two scars, and 13.2 +/- 5.3 for three scars . The second tuberculin reaction (PPD2), performed 2 wk later on the contralateral forearm, showed a marked increase in reactivity . The increase in reaction size was most evident in students who had BCG scars but who were initially PPD negative (less than 10 mm) . Smaller increases were observed in students without BCG scars, and also in those who had BCG scars but who were initially tuberculin positive (greater than or equal to 10 mm) . The persistence of the booster effect was evaluated by performing PPD3 1 yr later . PPD1-negative students with BCG scars maintained the increased level of reactivity to PPD2 after 1 yr . An immunizing effect of tuberculin testing was suggested in 11 nonimmunized students who were initially PPD negative.(ABSTRACT TRUNCATED AT 250 WORDS)

Infect Immun, 1990 Jul, 58(7), 2098 - 104
The 75-kilodalton cytoplasmic Chlamydia trachomatis L2 polypeptide is a DnaK-like protein; Birkelund S et al.; The gene coding for the 75-kilodalton cytoplasmic Chlamydia trachomatis L2 polypeptide has been cloned in Escherichia coli, and the nucleotide sequence has been determined . The cloned DNA fragment contained the coding region as well as the putative promoter . The deduced amino acid sequence of the 1,980-base-pair open reading frame revealed 94% homology with a 75-kilodalton protein from C . trachomatis serovar D and 57% homology with the DnaK proteins of E . coli and of Bacillus megaterium, while amino acid homology with human heat shock protein 70 (hsp70) was 42% . The promoter region was identified by computer search and by primer extension of mRNA synthesized in recombinant E . coli . The promoter region which differed from the putative promoter region in serovar D was shown to be a mixed promoter type in which the -10 region showed a regular TATA box configuration while the -35 region showed high homology with heat shock promoters . This mixed promoter was recognized in E . coli.

J Bacteriol, 1990 Jul, 172(7), 4017 - 22
Chitinase system of Bacillus circulans WL-12 and importance of chitinase A1 in chitin degradation; Watanabe T et al.; Bacillus circulans WL-12, isolated as a yeast cell wall-lytic bacterium, secretes a variety of polysaccharide-degrading enzymes into culture medium . When chitinases of the bacterium were induced with chitin, six distinct chitinase molecules were detected in the culture supernatant . These chitinases (A1, A2, B1, B2, C, and D) showed the following distinct sizes and isoelectric points: Mr 74,000, pI 4.7 (A1); Mr 69,000, pI 4.5 (A2); Mr 38,000, pI 6.6 (B1); Mr 38,000, pI 5.9 (B2); Mr 39,000, pI 8.5 (C); and Mr 52,000, pI 5.2 (D) . Among these chitinases, A1 and A2 had the highest colloidal-chitin-hydrolyzing activities . Chitinase A1 showed a strong affinity to insoluble substrate chitin . Purified chitinase A1 released predominantly chitobiose {(GlcNAc)2} and a trace amount of N-acetylglucosamine (GlcNAc) from colloidal chitin . N-terminal amino acid sequence analysis of chitinases A1 and A2 indicated that chitinase A2 was generated from chitinase A1, presumably by proteolytic removal of a C-terminal portion of chitinase A1 . Since chitinase A2 did not have the ability to bind to chitin, the importance of the C-terminal region of chitinase A1 to the strong affinity of chitinase A1 to substrate chitin was suggested . Strong affinity of the chitinase seemed to be required for complete degradation of insoluble substrate chitin . From these results, it was concluded that chitinase A1 is the key enzyme in the chitinase system of this bacterium.

J Urol, 1990 Jul, 144(1), 53 - 8
Effects of local bacillus Calmette-Guerin therapy in patients with bladder carcinoma on immunocompetent cells of the bladder wall; Bohle A et al.; The antitumoral effects of intravesical bacillus Calmette-Guerin against recurrent superficial urothelial bladder cancer seem to be linked to immunological effector mechanisms . To characterize further the local cellular response in the bladder wall of patients receiving intravesical bacillus Calmette-Guerin bladder biopsies were examined before and immediately after a 6-week course of bacillus Calmette-Guerin and for up to 1 year at 3-month intervals thereafter . The results showed a marked infiltration after bacillus Calmette-Guerin therapy predominantly of the suburothelial tissue with mononuclear cells, which expressed activation markers and accumulated to follicle-like granulomatous structures . With regard to T lymphocyte subsets the ratio of T helper-to-T suppressor cells in the bladder wall increased from 0.5 to nearly 2 . This phenomenon persisted even in biopsies 1 year after initial treatment . In some probes activated mononuclear cells invaded the urothelium, emanating from suburothelial granulomas . It is tempting to assume that these changes within the local immunocompetent cells are related to the therapeutic effect of bacillus Calmette-Guerin against bladder cancer.

Am J Public Health, 1990 Jul, 80(7), 848 - 52
Public health implications of the microbial pesticide Bacillus thuringiensis: an epidemiological study, Oregon, 1985-86; Green M et al.; Bacillus thuringiensis var . kurstaki (B.t.-k) is a microbial pesticide which has been widely used for over 30 years . Its safety for a human population living in sprayed areas has never been tested . Surveillance for human infections caused by B.t.-k among Lane County, Oregon residents was conducted during two seasons of aerial B.t.-k spraying for gypsy moth control . Bacillus isolates from cultures obtained for routine clinical purposes were tested for presence of Bacillus thuringiensis (B.t.) . Detailed clinical information was obtained for all B.t.-positive patients . About 80,000 people lived in the first year's spray area, and 40,000 in the second year's area . A total of 55 B.t.-positive cultures were identified . The cultures had been taken from 18 different body sites or fluids . Fifty-two (95 percent) of the B.t . isolates were assessed to be probable contaminants and not the cause of clinical illness . For three patients, B.t . could neither be ruled in nor out as a pathogen . Each of these three B.t.-positive patients had preexisting medical problems . The level of risk for B.t.-k and other existing or future microbial pesticides in immunocompromised hosts deserves further study.

Cancer Res, 1990 Jul 1, 50(13), 4032 - 7
Induction of interleukin 3 and tumor resistance by SSM, a cancer immunotherapeutic agent extracted from Mycobacterium tuberculosis; Sasaki H et al.; Interleukin 3 (IL-3) activity was demonstrated when inguinal lymph node cells obtained from Bacillus Calmette-Guerin-sensitized mice (BCG-ILNC) were stimulated in vitro with SSM, an immunomodulator extracted from Mycobacterium tuberculosis . The IL-3 activity was first detected on Day 1 in culture fluids of BCG-ILNC stimulated with SSM, reached a peak on Day 3, and then gradually decreased . The activity was completely neutralized by treatment with anti-murine IL-3 monoclonal antibody (mAb) . When BCG-ILNC were treated with anti-Thy 1.2 or anti-Lyt 1.2 mAb followed by complement, IL-3 was not produced in the culture fluids . However, IL-3 in the culture fluids was detected when BCG-ILNC were treated with anti-Lyt 2.2 mAb, anti-asialo-GM1, or anti-mouse immunoglobulin antiserum followed by complement . These results suggested that Lyt 1+ T-cells appeared to be required for the production of IL-3 from BCG-ILNC stimulated with SSM . In addition, low but significant IL-3 activity was also observed in sera of mice treated with SSM . However, serum IL-3 activity was not detected in mice treated with both SSM and Thy 1.2 or Lyt 1.2 mAb, whereas the activity was induced by SSM in mice treated with anti-Lyt 2.2 mAb or anti-asialo-GM1 antiserum . On the other hand, the in vivo growth of IMC tumors inoculated in BALB/c x DBA/2 F1 mice was significantly decreased by intralesional injection of culture fluids containing IL-3, as well as by SSM itself . This antitumor activity of the culture fluids was not altered when it was treated with mAbs for interleukin 1, interleukin 2, or anti-mouse gamma-interferon antiserum . The antitumor activity of the fluid was only eliminated when it was treated with anti-mouse IL-3 mAb . Since nonspecific resistance to tumors in mice stimulated with SSM appears to require Lyt 1+ T-cells, these results suggest that, in part, nonspecific resistance to tumors of mice stimulated with SSM may be developed through IL-3, which was produced by Lyt 1+ T-cells after SSM stimulation.

Proc Natl Sci Counc Repub China B, 1990 Jul, 14(3), 175 - 82
Characterization of the Bacillus thuringiensis strains isolated from Taiwan; Chak KF et al.; Over 100 Bacillus thuringiensis (Bt) isolates which produced phase bright inclusions have been isolated from soil samples from different areas in Taiwan . Three types of crystal proteins were visualized by phase contrast microscopy . Among these isolates, only 14 different types of plasmid profiles have been observed . They all possess a variety of plasmids ranging from a few kb to around 250 kb in size . With respect to the crystal protein profiles, the plasmid profiles, and the shapes of crystal proteins, we found that the majority of our isolates (87%) were different from most of the known Bt strains . Our other two types of isolates (10 and 3%) resembled Bt var . kurstaki HD1 and Bt var . israelensis, respectively . Most of our isolates were active against Bombyx mori (Lepidoptera) and Aedes aegypti (Diptera) . Most interestingly, two of our isolates, Nos . 82 and 96, were found highly toxic to Heliothis virescens, even compared with the standard strain, Bt var . kurstaki HD1 . Using insecticidal crystal protein (ICP) gene probe from Bt var . aizawai HD-133 to probe the total DNA of our isolates, we observed that at least one plasmid from each of the tested strains reacted with the probe . A 10 kb plasmid from some of our isolates hybridized with the probe . This probably is the first evidence demonstrating that the ICP gene sequence can be found in a low molecular weight plasmid.

Mol Gen Genet, 1990 Jul, 222(2-3), 278 - 83
Structure of the beta-1,3-1,4-glucanase gene of Bacillus macerans: homologies to other beta-glucanases; Borriss R et al.; The nucleotide sequence of an 852 base pair (bp) DNA fragment containing the entire gene coding for thermostable beta-1,3-1,4-glucanase of Bacillus macerans has been determined . The bglM gene comprises an open reading frame (ORF) of 711 bp (237 codons) starting with ATG at position 93 and extending to the translational stop codon TAA at position 804 . The deduced amino acid sequence of the mature protein shows 70% homology to published sequences of mesophilic beta-1,3-1,4-glucanases from B . subtilis and B . amyloliquefaciens . The sequence coding for mature beta-glucanase is preceded by a putative signal peptide of 25 amino acid residues, and a sequence resembling a ribosome-binding site (GGAGG) before the initiation codon . By contrast with the processed protein, the N-terminal amino acid sequence constituting the putative leader peptide bears no or only weak homology to signal peptides of mesophilic Bacillus endo-beta-glucanases . The B . macerans signal peptide appears to be functional in exporting the enzyme to the periplasm in E . coli . More than 50% of the whole glucanase activity was localized in the periplasmic space and in the supernatant . Whereas homology to endo-1,4-beta-glucanases is completely lacking, a weak amino acid homology between the sequence surrounding the active site of phage T4 lysozyme and a sequence spanning residues 126 through 161 of B . macerans endo-beta-glucanase could be identified.

Ann Oncol, 1990 Jul, 1(4), 255 - 61
Empiric treatment of infection during granulocytopenia; Klastersky J; Results from clinical trials conducted over the past 15 years suggest the following: a) Early empiric therapy with broad-spectrum antibiotics directed against Gram-negative bacillary bacteremia is necessary in febrile granulocytopenic cancer patients; b) The level and dynamics of the granulocyte count are extremely important in determining the outcome of bacteremia; c) Most empiric antimicrobial regimens will require therapeutic modifications; these alterations are necessary and contribute to a high overall success rate; d) Only microbiologically documented infections and especially bacteremias are useful for comparison of initial response to antimicrobial regimens; e) The response rate of Gram-negative bacillary bacteremia is clearly influenced by the susceptibility of the causative pathogen to the beta-lactam component of the empiric regimen; emergence of resistance to some beta-lactam antibiotics is quite common and necessitates successive modifications of empiric regimens with time; f) The combination of an anti-pseudomonal beta-lactam with an aminoglycoside is recommended as the standard for empiric therapy in febrile granulocytopenic cancer patients, especially in those with severe and persistent granulocytopenia who are suspected of having Gram-negative bacillary bacteremia; less neutropenic and/or asymptomatic patients may do well with monotherapy; g) Gram-positive pathogens have become a common cause of bacteremia in granulocytopenic cancer patients; the response rate to empiric regimens may be suboptimal but the associated mortality is low; h) Patients with severe granulocytopenia and protracted fever whose blood cultures remain negative are at high risk for contracting fungal infections; in these patients, empiric antifungal agents are probably indicated.

Indian J Lepr, 1990 Jul-Sep, 62(3), 351 - 7
Is leprosy bacillus a chemo-autotrophic nocardioform organism?
Pal D, Chakrabarty AN, Dastidar SG.
Numerous attempts at in vitro cultivation of the leprosy bacillus have all proved to be unsuccessful . Recently, we have repeatedly isolated chemo-autotrophic nocardioform (CAN) organisms in pure culture from multibacillary cases of leprosy . We find that these resemble the leprosy bacillus in many respects and suggest that the leprosy bacillus may be closer to the genus Nocardia than to Mycobacterium, and that it may be a chemo-autotroph, requiring only simple sources of carbon and nitrogen for its growth . This is in contrast to most other human pathogens, which are heterotrophs requiring complex sources of carbon and nitrogen for their growth . This could offer a possible explanation for the repeated failure at in vitro cultivation of the leprosy bacillus.

Indian J Lepr, 1990 Jul-Sep, 62(3), 310 - 5
Assessment of testicular volume in bacilliferous leprosy: correlation with clinical parameters; Abraham A et al.; Testicular involvement in leprosy was studied in 30 multi-bacillary (BL/LL) patients . Ten (33.3%) gave past history of type II reactions, of whom nine (30%) gave history of testicular pain and/or swelling . Decreased libido was a common complaint (63.3%) . Gynaecomastia was noted in 3 patients (10%) and altered hair pattern in 11 patients (36.7%) . Testicular sensation was impaired in 10 (33.3%) patients . Testicular volume was assessed objectively using the Prader orchidometer and found to be reduced in nine (30%) patients . Reduction in testicular volume correlated with longer duration of disease and a past history of type II reaction.

Avian Dis, 1990 Jul-Sep, 34(3), 779 - 86
Characterization of a gram-positive bacterium from the proventriculus of budgerigars (Melopsittacus undulatus); Scanlan CM et al.; The cellular, cultural, and biochemical characteristics of eight isolates of a large gram-positive bacillus that are commonly observed as apparently normal flora in the proventriculus of budgerigars (Melopsittacus undulatus) were determined . The bacterium was highly pleomorphic and changed markedly in both diameter and length when subcultured on agar media . The bacterium was facultative anaerobic and capnophilic, hemolytic on blood agar, and formed flat colonies with irregular edges after incubation for several days . All isolates grew on sodium azide agar but did not grow on MacConkey agar . The isolates were catalase-negative and oxidase-negative and did not reduce nitrate . All isolates failed to utilize arginine, lysine, ornithine or tryptophane but produced acid from glucose, galactose, levulose, maltose, melibiose, starch, and sucrose . All isolates produced acetoin from glucose and hydrolyzed esculin . The eight isolates could not be identified to either genus or species level based on the descriptions of currently classified organisms in the division Firmicutes as described in Bergey's Manual of Systematic Bacteriology.

Prikl Biokhim Mikrobiol, 1990 Jul-Aug, 26(4), 528 - 33
{Characteristics of hydrolase biosynthesis in Bacillus mesentericus grown on various media}; Kovalenko EA et al.; The dynamics of the consumption of major carbon and nitrogen sources and the biosynthesis of hydrolytic enzymes were studied in Bacillus mesentericus grown on semisynthetic media . Conditions were chosen that provide the obtaining of the culture liquid with predominantly proteolytic or amylolytic activity . The replacement of maltose with native starch resulted in more intensive accumulation of the biomass and hydrolytic enzymes, and in more rapid (by 3-5 hr) transformation from the logarithmic to the stationary growth phase.

J Gen Microbiol, 1990 Jul, 136 ( Pt 7), 1327 - 34
Molecular cloning and nucleotide sequence of a gene for alkaline cellulase from Bacillus sp . KSM-635; Ozaki K et al.; A gene for alkaline cellulase from the alkalophilic Bacillus sp . KSM-635 was cloned into the HindIII site of pBR322 and expressed in Escherichia coli HB101 . Although the recombinant plasmid contained two HindIII inserts of 2.6 kb and 4.0 kb, the inserts were found to be contiguous in the Bacillus genome by hybridization analysis . Nucleotide sequences of a 2.4 kb region which was indispensable for the production of cellulase, and the flanking, 1.1 kb region, were determined . There was an open reading frame (ORF) of 2823 bp in the 3498 bp sequence determined, which encoded 941 amino acid residues . Two putative ribosome-binding sites and a sigma 43-type, promoter-like sequence were found upstream from an initiation codon in the ORF . The deduced amino-terminal sequence resembles the signal peptide of extracellular proteins . A region of amino acids, 249 to 568, of the deduced amino acid sequence of the cellulase from this organism is homologous with those of alkaline and neutral enzymes of other micro-organisms, but nine amino acid residues were found to be conserved only in the alkaline enzymes.

FEMS Microbiol Lett, 1990 Jul, 58(2), 131 - 5
Nucleotide sequence of the G6-amylase gene from alkalophilic Bacillus sp . H-167; Shirokizawa O et al.; The nucleotide sequence of the G6-amylase gene from alkalophilic Bacillus sp . H-167 was determined . The open reading frame of the gene consisted of 2865 base pairs, encoding 955 amino acids . The NH2-terminal amino acid sequence analysis of the G6-amylase indicated that the enzyme had a single peptide of 33 amino acid residues and the mature enzyme was composed of 922 amino acids, giving a molecular mass of 102,598 . Identity of the NH2-terminal amino acid sequences among each component of the multiform G6-amylase suggested the proteolytic processing of the COOH-terminal side of the enzyme . The DNA sequence and the deduced amino acid sequence of the G6-amylase gene showed no homology with those of other bacterial alpha-amylases although the consensus amino acid sequences of the active center were well conserved.

Clin Ther, 1990 Jul-Aug, 12(4), 352 - 6
Evaluation of IPM/CS in gram-negative bacillus infections: use of an endotoxin-specific assay; Endo S et al.; The subjects were 26 patients hospitalized with severe trauma or burns . Twice daily for seven to 14 days, 1 gm of IPM/CS (a 1:1 combination of imipenem and cilastatin sodium) was administered by intravenous drip infusion to each patient . Clinical outcome was rated excellent in nine patients, good in 12, and fair in five . Bacteria, isolated in 21 of the 26 patients, were eliminated in 18 and decreased in three . Mean endotoxin levels, assessed by the Endospecy method, were reduced from 167.8 pg/ml before treatment to 5.1 pg/ml after treatment . No treatment side effects were noted.

Biol Chem Hoppe Seyler, 1990 Jul, 371(7), 631 - 6
Nucleotide sequences of Bacillus stearothermophilus ribosomal protein genes: part of the ribosomal S10 operon; Kromer WJ et al.; Restriction fragments from Bacillus stearothermophilus chromosomal DNA were cross-hybridized with the Escherichia coli ribosomal protein L2 gene rplB . A 2-kb EcoRI fragment which showed cross-hybridization was cloned into the M13 phage and sequenced by the dideoxy chain-terminating method . Comparison of the deduced amino-acid sequences with the corresponding sequences of E . coli ribosomal proteins showed that this fragment contains the region encoding the C-terminus of L2, the genes encoding S19, L22, S3 as well as the N-terminus of L16 . Thus the organization of this gene cluster is the same as that in the S10 operon of E . coli . The deduced sequences of proteins L22 and S3, which have not been determined so far, were found to have 52% or 55% amino-acid identity, respectively, with those of the corresponding proteins in E . coli . The deduced B . stearothermophilus S19 protein sequence was in accordance with the reinvestigated protein sequence (H . Hirano, personal communication).

Nippon Hinyokika Gakkai Zasshi, 1990 Jul, 81(7), 997 - 1001
{Intravesical bacillus Calmette-Guerin (BCG) in the treatment of superficial bladder cancer . Prospective randomized study for prophylactic effect}; Yamamoto T et al.; We report the results of prospective randomized study which was designed to evaluate prophylactic effects of intravesical bacillus Calmette-Guerin (BCG) in the treatment of superficial transitional cell carcinoma of the bladder . A total of 44 cases who had no previous treatment of bladder cancer were randomly assigned to BCG or control groups after TUR . BCG group (23 cases) received intravesical instillation of 80 mg BCG (Tokyo strain) at one week intervals for 6 weeks, at two week intervals for 12 weeks, and at one month intervals for 20 months . In BCG groups, 3 cases had recurrence at 6 months and 1 case at 9 months, while the other 19 cases had no recurrent disease for 3 to 34 months (average 20.3 months) of follow-up . Control group (21 cases) had no further treatment after TUR . In control group, recurrence was seen at 3 months in 3 cases, at 6 months in 5 cases, at 9 months in 2 cases, at 12 months in 3 cases and at 21 months in 1 case . Only 7 cases in the control group were free of recurrence for 8 to 45 months (average 32.3 months) of follow-up . One and two year-recurrence rates in BCG group (18.4%, 18.4%) was significantly (p less than 0.01) lower than those in control group (63.2%, 68.9%) . Among the complications of intravesical BCG were cystitis (76.2%), low grade fever (13.0%), and several days' persistent gross hematuria (13.0%) . Most of these signs were self-limited and only in 2 cases instillation of BCG was discontinued.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Pediatr, 1990 Jul, 149(10), 700 - 4
Bare lymphocyte syndrome--combined immunodeficiency and neutrophil dysfunction; Will N et al.; A 4-year-old girl presented with recurrent infections . Immunoglobulin deficiency (serum and secretory IgA, serum IgG3) neutropenia and neutrophil dysfunction (defective spontaneous migration and chemotaxis) were found . T-lymphocyte counts were normal and they responded to phytohaemagglutinin but were not stimulated by Concanavalin A, pokeweed mitogen and microbial antigens in vitro . Delayed cutaneous hypersensitivity testing to purified protein derivative and candidin was negative . Despite bacille Calmette-Guerm vaccination and candidiasis, near normal beta-2-micro-globulin and human leucocyte antigen (HLA) class I concentrations were detected on mononuclear cells and phytohaemagglutinin-induced lymphoblasts . HLA class II antigens (HLA-DP, -DQ, -DR) were not expressed . These observations indicated a bare lymphocyte syndrome (BLS) type II . This is the first time neutrophil dysfunction has been noted in association with BLS.

Zhonghua Yu Fang Yi Xue Za Zhi, 1990 Jul, 24(4), 199 - 202
{A methodological study on treatment of hospital sewage}; Zhou YL; With the view to finding a more effective and economic system for the disposal of hospital sewage, a series of experimental and on the spot investigations were conducted . The results are as follows . Disinfection must be taken as the key link in the treatment of hospital sewage . After primary treatment and chlorination, when the product value of concentration (mg/L) multiplied by time (min.) achieved to 240 (general hospital) and 540 (tuberculosis hospital), the content of suspended substances (SS) fell to 37 mg/L; the clearance rate of SS reached 82% . The BOD5 fell to 35 mg/L a drop to 42% . E . coli was less than 9 individual/L, the killing rate reached 99.999 99% . The intestinal pathogens and tubercle bacillus were completely wiped out . The sludge from the sewage can be treated with lime {Ca(OH)2} and when the pH value rose to 12, the requirement of disinfection was satisfied and both the sludge and sewage can be drained . The aeration of sewage through shooting flow, biological oxidation combined with sediment action by passing thru reclining tubes is an effective way for the secondary treatment of hospital sewage . In addition, we developed the double siphon equipment with water power-automatic controller, the WD-700 flowmeter, the anti-corrosive paint coating the contact pond; according to test parameters, we designed a simplified evaluation graph for the purpose of surveillance.

Am J Clin Pathol, 1990 Jul, 94(1), 80 - 3
The risk of bacterial growth in units of blood that have warmed to more than 10 degrees C; Saxena S et al.; Most transfusion services discard unopened units of blood that have been returned to the blood bank more than 30 minutes after the issuance or have attained a temperature of more than 10 degrees C . The objective of this study was to learn the prevalence of bacterial growth, if any, in the units of blood that were exposed twice, for six hours each time, to room temperature during their refrigerated storage . All of the 396 units cultured were negative except one red cell unit that grew a Bacillus species, probably B . subtilis . Further studies suggested that the growth of B . subtilis was due to laboratory contamination . The authors concluded that more work is needed to study the bacterial growth and the effect on red cell enzymes in the units of blood that are exposed to room temperature for varying periods or are returned to the blood bank after 30 minutes of issuance . If no adverse effect is noted, the policy of not reissuing such units may need revision so that more units could be salvaged.

J Urol, 1990 Jul, 144(1), 65 - 7
A randomized prospective comparison of oral versus intravesical and percutaneous bacillus Calmette-Guerin for superficial bladder cancer; Lamm DL et al.; Reports of a dramatic decrease in tumor recurrence and regression of muscle invasive disease with oral bacillus Calmette-Guerin prompted us to conduct a randomized prospective comparison of oral bacillus Calmette-Guerin with the standard intravesical plus percutaneous therapy . Oral therapy consisted of 200 mg . Tice bacillus Calmette-Guerin 3 times each week . Intravesical and percutaneous Tice bacillus Calmette-Guerin at a dose of 50 mg . was given weekly for 6 weeks, at 8, 10 and 12 weeks, then at 6 months and every 6 months thereafter . Minimal side effects confirmed the safety of oral bacillus Calmette-Guerin . Tumor recurrence was documented in 21 of 33 oral bacillus Calmette-Guerin patients (64%) and 18 of 55 (33%) who received intravesical plus percutaneous therapy (p less than 0.002, chi-square test) . We were unable to demonstrate any antitumor activity of oral bacillus Calmette-Guerin in this study.

Cancer Res, 1990 Jul 1, 50(13), 3843 - 7
Comparison of the fibronectin-binding ability and antitumor efficacy of various mycobacteria; Hudson MA et al.; Although the mechanism by which Bacillus Calmette-Guerin (BCG) exerts an antitumor effect on superficial bladder tumors is not fully understood, recent evidence has implicated binding of BCG organisms to fibronectin (FN) as requisite for this antitumor efficacy . Various substrains of BCG and other mycobacteria were tested in vitro for their relative capacities to bind both matrix and soluble FN . A substrain of Mycobacterium kansasii, designated the "high-binding strain," was found to bind FN more readily (P less than 0.05) in in vitro studies, when compared to commercially available substrains of BCG (Tice, Connaught, and Armand Frappier) . The binding by the three commercial strains of BCG to FN in vitro appeared to be equivalent . The high-binding strain was further demonstrated to attach more readily in vivo to the acutely injured murine bladder (P less than 0.005) than the Armand Frappier substrain . Finally, using the MB49 murine bladder tumor model, an enhanced antitumor effect (P less than 0.05) was noted in mice treated with intravesical high-binding strain, in comparison to the Armand Frappier substrain, during five weekly treatments . It appears not only that the commercial substrains of BCG bind FN in an equivalent manner but also that the relative binding capacities of the substrains correlate directly with antitumor activity . A substrain of M . kansasii appears to have been identified which may prove more clinically effective than the currently available strains of BCG.

Med Trop (Mars), 1990 Jul-Sep, 50(3), 307 - 9
{Anatomo-clinical forms of skin tuberculosis . (Dermatology service-Regional Hospital Center of Bouake, Ivory Coast)}; Darie H et al.; Tuberculosis of the skin is not exceptional in Africa South of the Sahara . The different ways of contamination of the skin by the tubercular bacillus and the level of immunologic reaction of the host account for the clinical polymorphisms . The bacillus is detected with difficulty, and the classical anatomo-pathological feature of tuberculoid granuloma is rarely complete: A classification based on physiopathological, clinical, bacteriological and histological criteria is an help to diagnosis.

J Invertebr Pathol, 1990 Jul, 56(1), 86 - 90
Light microscope immunolocation of Bacillus thuringiensis kurstaki delta-endotoxin in the midgut and Malpighian tubules of the tobacco budworm, Heliothis virescens; Ryerse JS et al.; Light microscope immunofluorescence was used to localize the membrane binding of Bacillus thuringiensis kurstaki 63-kDa delta-endotoxin in Heliothis virescens midgut and Malpighian tubules . Staining was observed along all exposed mucosal (apical microvillar) plasma membranes . Interpretation of the serosal (basal) plasma membrane staining was complicated because the basal lamina also stained . The results suggest that the toxin binds to all exposed plasma membranes without apparent specificity for particular membrane domains.

J Gen Virol, 1990 Jul, 71 ( Pt 7), 1535 - 44
Construction of genetically engineered baculovirus insecticides containing the Bacillus thuringiensis subsp . kurstaki HD-73 delta endotoxin; Merryweather AT et al.; The delta-endotoxin gene from Bacillus thuringiensis subsp . kurstaki HD-73 was inserted into Autographa californica nuclear polyhedrosis virus (AcMNPV) using two transfer vector systems . In the first, the delta-endotoxin gene was placed under the control of the polyhedrin gene promoter in lieu of the polyhedrin coding sequences, thus deriving a polyhedrin-negative virus . In the second, it was inserted under the control of a copy of the AcMNPV p10 promoter positioned upstream of the polyhedrin gene to produce a polyhedrin-positive virus . Analysis of infected cell extracts showed that the delta-endotoxin was expressed in insect cells as 130K, 62K and 44K proteins, with peak syntheses at 18 h post-infection . Each of these products reacted with antisera specific for the complete protoxin and the cleaved, active form . When extracts from the cells infected with the polyhedrin-negative virus were fed to Trichoplusia ni larvae, feeding by the insects was inhibited and deaths occurred that were inconsistent with virus infection . This effect was also observed after the inoculum had been treated with detergents to inactivate virus particles prior to feeding to the larvae . These data indicate that the expression of the B . thuringiensis delta-endotoxin gene by a baculovirus in insect cells produces material with insecticidal activity . The biological activities of the two recombinant viruses were assessed in conventional bioassay tests by feeding virus particles or occlusion bodies to the insects . The polyhedrin-negative virus preparation appeared to be contaminated with endotoxin which inhibited feeding of the insects and prevented determination of the LD50 value . The polyhedrin-positive virus had an LD50 value about twofold higher than that of unmodified AcMNPV . The significance of these data for the genetic engineering of virus insecticides is discussed.

J Bacteriol, 1990 Jul, 172(7), 3821 - 5
Physical map of the Bacillus cereus chromosome; Kolsto AB et al.; A physical map of the Bacillus cereus chromosome has been constructed by aligning 11 NotI fragments, ranging in size from 200 to 1,300 kilobases . The size of the chromosome is about 5.7 megabases . This is the first Bacillus genome of which a complete physical map has been described.

Am J Respir Cell Mol Biol, 1990 Jul, 3(1), 3 - 10
The role of the type 3 complement receptor in the induced recruitment of myelomonocytic cells to inflammatory sites in the mouse; Rosen H et al.; The type 3 complement receptor (CR3), initially identified as the leukocyte cell surface receptor for iC3b, is now known to form part of the extended integrin family of cell adhesion molecules that mediate both cell-cell and cell-extracellular matrix interactions . The identification of a heritable deficiency of human leukocyte adhesion together with the advent of monoclonal antibodies has shed some light on the central role of CR3 in the transendothelial migration of macrophages and neutrophils to sites of inflammation . We review the general structural features of CR3 and then examine our understanding of its role in both nonspecific and T cell-dependent inflammatory processes based on our murine in vivo experiments . CR3-dependent inflammation seems to contribute to the pulmonary response to some stimuli (lipopolysaccharide) but not to others (bacillus Calmette-Guerin) . These studies highlight the potential therapeutic benefits, as well as the significant risks of potentiating acute bacterial infections, of CR3 blockade in vivo.

Med Vet Entomol, 1990 Jul, 4(3), 283 - 8
Bait-fed adult Culex pipiens carry the larvicide Bacillus sphaericus to the larval habitat; Schlein Y et al.; Sugar meals of plant origin are a basic component of mosquito diet . We show that sugar baits have potential as vehicles for control agents . When Culex pipiens L . resting-sites were sprayed with sucrose solution, with or without incorporation of the larval toxicant Bacillus sphaericus Neide, 47% of female mosquitoes fed in situ . Dispersing B . sphaericus-carrier mosquitoes caused larval mortality in breeding-sites 60-100 m from the sprayed resting-sites . The effect was not seen where no adults rested in sparse vegetation above larval habitats . This approach may be useful for the application of biological control agents against mosquitoes in biotopes where the adults and larvae are juxtaposed.

J Gen Microbiol, 1990 Jul, 136 ( Pt 7), 1381 - 5
Growth of Bacillus stearothermophilus on glycerol in chemostat culture: expression of an unusual phenotype; Burke RM et al.; Bacillus stearothermophilus grew readily on glycerol in carbon-limited chemostat culture and expressed a high carbon conversion efficiency . However, the strain of organism used (probably B . stearothermophilus var . nondiastaticus) proved particularly sensitive to glycerol, both respiration and growth being severely impeded by any surfeit of this compound . Sensitivity was found to correlate with an exceptionally high level of expression of glycerol kinase {activities of more than 80 mumol min-1 (mg protein)-1 were manifest in crude cell-free extracts}, coupled with low activities of methylglyoxal synthase and of glyoxylase (enzymes of the methylglyoxal bypass) . It is proposed that metabolic dysfunction results from an uncontrolled gross accumulation of glycerol phosphate (and early products of its metabolism) within the cells, coupled with depletion of the intracellular phosphate pool.

J Appl Bacteriol, 1990 Jul, 69(1), 73 - 9
Psychrotrophic strains of Bacillus cereus producing enterotoxin; van Netten P et al.; In investigations on three outbreaks of Bacillus cereus food poisoning in Spain and The Netherlands, the causative strains grew within a temperature range of 4-37 degrees C, but not at 43 degrees C . Such psychrotrophic types were found to occur in various dairy products (including ca 25% of 35 samples of pasteurized milk) and some mousses and cook/chill meals . Growth of and enterotoxin production by psychrotrophic B . cereus could be prevented by temperatures below 4 degrees C and pH-values not exceeding 5.0.

Minerva Med, 1990 Jul-Aug, 81(7-8), 547 - 53
{The current epidemiologic profile of the drug resistance of Koch's bacillus to antitubercular antibiotic chemicals}; Giove S et al.; The study was conducted on patients with initially diagnosed tuberculosis encountered in 1986-87 . A series of 269 bacterial antibiograms performed on sputum {correction of excreate} and other materials was examined in order to ascertain the resistance of Koch's bacillus to R/AMP, INH, SM and EMB . Primary drug resistance was found in 64 cases (23.79% of the total) and was isolated in 32 (11.89%), multiple in the rest . The response to the individual antibiotics was a follows resistance to INH was isolated in 11 cases (4.08%), combined with resistance to other drugs in 27 (10.03%); resistance to SM was isolated in 13 cases (4.81%) multiple in 24 (8.92%); resistance to R/AMP was isolated in 4 cases (1.48%), multiple in 14 (5.2%); resistance to EMB was isolated in 4 (1.48%) multiple in 19 (7.06%) . Comparison with earlier studies in the Turin area highlights that the incidence of TB, in decline up to 1979, significantly increased in the period considered.

Infect Immun, 1990 Jul, 58(7), 2220 - 7
A novel bicomponent hemolysin from Bacillus cereus; Beecher DJ et al.; A procedure combining isoelectric focusing (Sephadex IEF) and fast protein liquid chromatography (Superose 12; Mono-Q) removed hemolytic activity (presumably a contaminant) from partially purified preparations of the multicomponent diarrheal enterotoxin produced by Bacillus cereus . However, when the separated fractions were recombined, hemolytic activity was restored, suggesting that hemolysis is a property of the enterotoxin components . Combined fractions exhibited a unique ring pattern in gel diffusion assays in blood agar . During diffusion of the hemolysin from an agar well, the erythrocytes closest to the well were not lysed initially . After diffusion, hemolysis was observed as a sharp ring beginning several millimeters away from the edge of the well . With time the cells closer to the well were also lysed . This novel hemolysin consists of a protein (component B) which binds to or alters cells, allowing subsequent lysis by a second protein (component L) . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and Western blot analysis showed that hemolysin BL has properties similar to those described previously for the enterotoxin and that both components are distinct from cereolysin and cereolysin AB.

J Bacteriol, 1990 Jul, 172(7), 4032 - 6
Construction by site-directed mutagenesis of a 39-kilodalton mosquitocidal protein similar to the larva-processed toxin of Bacillus sphaericus 2362; Broadwell AH et al.; After ingestion of the parasporal crystals of Bacillus sphaericus, mosquito larvae process the 42-kilodalton (kDa) toxin to a protein of 39 kDa, which has an increased toxicity (A . H . Broadwell and P . Baumann, Appl . Environ . Microbiol . 53:1333-1337, 1987) . A similar activation is performed by trypsin and chymotrypsin . Using site-directed mutagenesis, we have constructed derivatives of the 42-kDa toxin with a deletion of 10 amino acids at the N terminus and deletions of 7, 17, or 20 amino acids at the C terminus . Toxicity for mosquito larvae was retained upon deletion of 7 or 17 amino acids but was lost upon deletion of 20 amino acids . Evidence is presented indicating that the protein containing deletions of 10 amino acids at the N terminus and 17 amino acids at the C terminus (corresponding to potential chymotrypsin cleavage sites) is similar to the 39-kDa protein produced in mosquito larvae or by digestion with chymotrypsin . Digestion with trypsin appears to generate a protein lacking 16 or 19 amino acids from the N terminus and 7 amino acids from the C terminus . As is the case with the recombinant-made 42-kDa protein, toxicity of its derivatives is dependent on the presence of a 51-kDa protein which is a component of the parasporal crystal of B . sphaericus 2362.

J Bacteriol, 1990 Jul, 172(7), 3940 - 5
Overproduction of tyrosyl-tRNA synthetase is toxic to Escherichia coli: a genetic analysis; Bedouelle H et al.; The tyrS genes from Escherichia coli and Bacillus stearothermophilus were toxic to E . coli when they were carried by plasmids with very high copy numbers (pEMBL8 and pEMBL9) . We quantified this effect by comparing the efficiencies of plating of E . coli derivatives harboring recombinant plasmids in various experimental conditions . The toxicity was apparent at both 30 and 37 degrees C . It increased with the growth temperature, the strength of the tyrS promoter, and the copy number of the plasmidic vector . Two- to threefold enhancement of tyrS expression raised the toxicity 300-fold . Point mutations in tyrS that prevent interaction between its product, tyrosyl-tRNA synthetase, and tRNA(Tyr) but do not alter the rate of formation of tyrosyl-adenylate abolished the toxicity . Thus, the toxic effect was due to high cellular levels of synthetase activity . At 30 degrees C, the cellular concentration of tyrosyl-tRNA synthetase reached 55% of that of soluble proteins and led to decreased beta-galactosidase stability . We discuss possible causes of this toxic effect and describe its applications to the study of the recognition and interaction between the synthetase and tRNA(Tyr).

Biochem Biophys Res Commun, 1990 Jun 29, 169(3), 1068 - 74
Dynamic structure of bacterial ribosomal 5S RNA helices II and III of B . megaterium 5S RNA; Kim JH et al.; A possible switch between two conformations, previously observed in an enzymatically cleaved fragment of E . coli 5S ribosomal RNA (a Gram-negative bacterium) containing helices II and III, has been examined by means of proton nuclear magnetic resonance spectroscopy (10-15 ppm) as a function of {Mg2+} and temperature for an RNase-T1 digested fragment of Bacillus megaterium 5S rRNA (a Gram-positive bacterium) containing the same helices II and III . The conformational changes induced in the fragment are not accompanied by breakage of some base-pairs and formation of others, but rather consist simply of tightening or loosening of helices with retention of existing base-pairs . Helix III is found to be more flexible than helix II . Finally, the loop conformation is conserved over a wide range of Mg2+ concentration, suggesting that the loop may serve an important role in the biological function of 5S rRNA in ribosomes.

Biochim Biophys Acta, 1990 Jun 28, 1045(1), 17 - 20
Composition of polar lipid acyl chains of Bacillus stearothermophilus as affected by temperature and calcium; Martins LO et al.; Bacillus stearothermophilus was grown within the temperature range of 48 to 68 degrees C in a complex medium and in the range of 45 to 72 degrees C in the presence of 2.5 mM Ca2+ . The main fatty acids of lipid extracts contain 15 to 17 carbon atoms, mostly branched-chain species . The most prevalent saturated straight-chain fatty acid is n-C16 . The total amount of branched-chain species decreases with increasing temperature of growth from 48 to 68 degrees C, whereas the straight-chain species increase . Thus, n-C16 almost doubles while i-C16, i-C17 and a-C17 decrease by 41.2, 28.9 and 41.9%, respectively . In the presence of Ca2+, the lipid metabolism favours the biosynthesis of straight-chain fatty acids with depression of branched-chain species, especially at lower temperatures . At high temperatures, Ca2+ has a less pronounced effect in the lipid biosynthesis . However, above 68 degrees C, a significant decrease is observed among the branched-chain fatty acids i-C15, i-C17 and a-C17 with a consequent increase in n-C16 . Furthermore, a remarkable increase is observed in oleic acid (from 2.7% at 68 degrees C to 11.5% at the extreme 72 degrees C.

Carbohydr Res, 1990 Jun 15, 201(1), 125 - 34
Isolation and characterization of three positional isomers of diglucosylcyclomaltoheptaose; Koizumi K et al.; Three positional isomers of diglucosylcyclomaltoheptaose {(G)2-beta-cyclodextrin}, 6(1),6(4)-di-O-(a-D-glucopyranosyl)-cyclomaltoheptaose (1), 6(1),6(3)-di-O-(a-D-glucopyranosyl)-cyclomaltoheptaose (2), and 6(1),6(2)-di-O-(a-D-glucopyranosyl)-cyclomaltoheptaose (3) were isolated by h.p.l.c . on a reversed-phase column from the mother liquors of a large-scale preparation of beta CD with Bacillus ohbensis cyclomaltodextrin glucanotransferase (EC 2.4.1.19) and were characterized by h.p.l.c . analysis of partial hydrolyzates and by 13C-n.m.r . spectroscopy . Their molecular weights were confirmed by f.a.b.-m.s . Their characteristic chromatographic behavior on four h.p.l.c . columns of different separation modes was found to be very useful for their identification . It is particularly noteworthy that the first application of a graphitized carbon column to CDs enabled a fine separation of all three positional isomers.

FEMS Microbiol Lett, 1990 Jun 15, 58(1), 107 - 13
Expression in Escherichia coli of the carboxy terminal domain of the BLAR sensory-transducer protein of Bacillus licheniformis as a water-soluble Mr 26,000 penicillin-binding protein; Joris B et al.; A cloning vector has been constructed which allows production and export by Escherichia coli of the Met346-Arg601 carboxy terminal domain of the 601 amino acid BLAR sensory-transducer involved in beta-lactamase inducibility in Bacillus licheniformis . The polypeptide, referred to as BLAR-CTD, accumulates in the periplasm of E . coli in the form of a water-soluble, Mr 26,000 penicillin-binding protein . These data and homology searches suggest that BLAR has a membrane topology similar to that of other sensory-transducers involved in chemotaxis.

Biochem J, 1990 Jun 15, 268(3), 671 - 7
Cloning of aminoglycoside phosphotransferase (APH) gene from antibiotic-producing strain of Bacillus circulans into a high-expression vector, pKK223-3 . Purification, properties and location of the enzyme; Sarwar M et al.; The aminoglycoside phosphotransferase gene from a butirosin-producing strain of Bacillus circulans was cloned in a high-expression vector (pKK223-3) to give the recombinant plasmid pMS5 . Escherichia coli harbouring the plasmid, E . coli JM103{pMS5}, was characterized, and several features of the expression of the phosphotransferase were studied . The phosphotransferase activity was best expressed in a medium lacking glucose, and the highest levels of the enzyme were found between 12 and 24 h of growth . The induction of the phosphotransferase expression with isopropyl beta-D-thiogalactopyranoside (inducer) was found to be undesirable as the overproduction of the enzyme led to the killing of the bacteria . The subcellular location of the phosphotransferase, and also the site in vivo of the phosphorylation of neomycin, was found to be in the cytoplasm . The phosphotransferase was purified to homogeneity in good yield (17 mg of purified protein/3 litres of culture) and was shown to be a monomer of Mr 30,000-32,000 . The N-terminal amino acid sequence was in agreement with that predicted from the gene sequence and confirmed the absence of any signal sequence . The regiospecificity of the phosphotransferase reaction was studied by m.s . and by 1H-, 13C- and 31P-n.m.r . using ribostamycin as the substrate, and it was found that the antibiotic was phosphorylated at the 3'-hydroxy group.

Cell, 1990 Jun 15, 61(6), 1113 - 20
Phospholipase C-mediated hydrolysis of phosphatidylcholine is an important step in PDGF-stimulated DNA synthesis; Larrodera P et al.; Recent evidence suggests the involvement of phosphatidylcholine (PC) hydrolysis both in the control of normal cell growth and in transformation . We show here that the simple exogenous addition of Bacillus cereus PC-hydrolyzing phospholipase C (PC-PLC) is sufficient to elicit a potent mitogenic response in Swiss 3T3 fibroblasts by a mechanism that is independent of protein kinase C . Our results on the additivity and synergism between B . cereus PC-PLC, PDGF, and insulin in the mitogenic response indicate that this novel phospholipid degradative pathway may be important in the mitogenic signaling cascade activated by PDGF.

Biochem Biophys Res Commun, 1990 Jun 15, 169(2), 765 - 72
Delta-endotoxins form cation-selective channels in planar lipid bilayers; Slatin SL et al.; Delta-endotoxins CryIA(c) and CryIIIA, two members of a large family of toxic proteins from Bacillus thuringiensis, were each allowed to interact with planar lipid bilayers and were analyzed for their ability to form ion-conducting channels . Both of these toxins made clearly resolved channels in the membranes and exhibited several conductance states, which ranged from 200 pS to about 4000 pS (in 300 mM KCl) . The channels formed by both toxins were highly cation-selective, but not ideally so . The permeability ratio of K+ to Cl- was about 25 for both channels . The ability of these proteins to form such channels may account for their toxic action on sensitive cells, and suggests that this family of toxins may act by a common mechanism.

J Immunol, 1990 Jun 15, 144(12), 4735 - 41
Expression of murine Fc receptors for IgG; Schreiber RE et al.; There are two distinct genes that encode murine low affinity Fc gamma RII, murine Fc gamma RII alpha, and murine Fc gamma RII beta, which are transcribed in specific cell lineages . Fc gamma RII alpha transcripts are present in macrophages, NK cells, and mesangial cells; Fc gamma RII beta transcripts are expressed in Fc gamma R-bearing B cells, T cells, and macrophages . We have devised a sandwich ELISA to quantify the expression of Fc gamma RII alpha protein . The ELISA is specific for Fc gamma RII alpha, and does not detect the closely related Fc gamma RII beta protein . Upon stimulation with IFN-gamma the Fc gamma RII beta- macrophage cell line J774a expressed a twelvefold enhanced level of Fc gamma RII alpha protein . Peritoneal macrophages synthesized varying amounts of Fc gamma RII alpha . High levels of Fc gamma RII alpha were observed in resident and thioglycollate-elicited peritoneal macrophages, but no Fc gamma RII alpha was detected in Bacillus Calmette Guerin-elicited macrophages . J774a cells stimulated with rIL-6 bound approximately twice as much anti-Fc gamma RII mAb 2.4G2 IgG as did unstimulated controls . However, the Fc gamma RII alpha-specific ELISA showed no change in the amount of Fc gamma RII alpha expressed . A probe encompassing the extracellular coding sequence of Fc gamma RII beta hybridized to two distinct transcripts that were elevated in rIL-6-stimulated J774a cells . One of these transcripts had the same mobility in electrophoresis as Fc gamma RII alpha mRNA and hybridized to an Fc gamma RII alpha-specific probe, whereas the other transcript was larger and did not hybridize to probes specific for either Fc gamma RII alpha or Fc gamma RII beta . Moreover, we confirmed, with an Fc gamma RII beta-specific probe, that J774a cells do not make Fc gamma RII beta mRNA . Thus, the larger transcript appears to encode a novel Fc gamma RII . We suggest that the increased level of binding of the anti-Fc gamma RII mAb 2.4G2 to rIL-6-induced cells represents translation of a Fc gamma R distinct from Fc gamma RII alpha or Fc gamma RII beta.

Biochemistry, 1990 Jun 12, 29(23), 5596 - 604
Rotational diffusion studies of the lipoyl domain of 2-oxoacid dehydrogenase multienzyme complexes; Harrison JP et al.; Rotational mobility of the lipoyl domain of a number of 2-oxoacid dehydrogenase complexes was investigated by transient dichroism after the domain had been specifically labeled with the triplet probe eosin-5-maleimide . Complexes investigated included pyruvate dehydrogenase complexes from Bacillus stearothermophilus, ox heart, and Escherichia coli (in which the E2 component had been genetically engineered to contain one lipoyl domain) and 2-oxoglutarate dehydrogenase complexes from ox heart and E . coli . Measurements were also performed with ox heart pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes specifically labeled on E1 . Anisotropy decays were recorded in glycerol-buffer solutions of varying viscosity and at different temperatures . For E2-labeled complexes, the decays were found to be multiexponential, and the fastest correlation time was considerably shorter than expected for tumbling of the whole complex . This fast correlation time was absent from E1-labeled complexes and was assigned to independent motion of the lipoyl domain . Plots of the fast correlation time against eta/T showed a surprisingly weak dependence on viscosity and extrapolated to a time of 30-40 microseconds at zero viscosity . To explain this result, a model is proposed in which the lipoyl domain is in equilibrium between "free" and bound states . The time of 30-40 microseconds is shown to correspond to 1/koff, where koff is the rate constant for dissociation of the domain from binding sites on the complex . This dissociation phenomenon only contributes to the anisotropy decay when the viscosity of the solution is sufficiently high to slow the tumbling of the whole complex to times that are long in comparison to 1/koff.

Appl Environ Microbiol, 1990 Jun, 56(6), 1608 - 14
Plasmid transfer between strains of Bacillus thuringiensis infecting Galleria mellonella and Spodoptera littoralis; Jarrett P et al.; To determine the possibility of plasmid transfer occurring between strains of Bacillus thuringiensis in infected lepidopterous larvae, Galleria mellonella and Spodoptera littoralis were infected with two or more strains of B . thuringiensis and the resulting bacteria from the dead insects were examined for plasmid transfer . Transfer rates of plasmids coding for crystal production and tetracycline resistance were high, reaching levels similar to those obtained in laboratory broth cultures . Transfer was higher in G . mellonella than S . littoralis, probably due to the greater ability of B . thuringiensis to colonize the larvae . In broth cultures, B . thuringiensis was also able to transfer plasmids into sporeforming bacteria present in soil samples . The results suggest that plasmid transfer between strains of B . thuringiensis occurs in nature, resulting in the production of new combinations of delta-endotoxins within populations of the bacteria.

J Am Mosq Control Assoc, 1990 Jun, 6(2), 300 - 5
Small scale field trials of Bacillus sphaericus (strain 2362) against anopheline and culicine mosquito larvae in southern Mexico; Arredondo-Jimenez JI et al.; Experimental breeding sites simulating natural conditions were used to evaluate the efficacy of 2 formulations of Bacillus sphaericus (strain 2362) against Anopheles albimanus and culicine (mostly Culex coronator and Cx . quinquefasciatus) mosquito larvae of southern Mexico . Three doses of each formulation were used in a first field trial: 2, 3 and 4 g/m2 (granular) or 2, 3 and 4 ml/m2 (liquid); and in a second field trial: 0.125, 0.24 and 0.5 g/m2 (granular) or 0.125, 0.25 and 0.5 ml/m2 (liquid) . The optimum concentrations of each formulation for effective control of larval populations over periods of 3-4 months were 0.125 ml/m2 of liquid product for Culex spp . and 2 g/m2 of granular product for An . albimanus (ca . 70% mean reduction).

Biotechnol Appl Biochem, 1990 Jun, 12(3), 245 - 51
Simple method for the isolation of the antilepidopteran toxin from Bacillus thuringiensis subsp . kurstaki; Venkateswerlu G et al.; A protein with a molecular mass of 66 kDa was isolated by a simple, rapid, and inexpensive method, using 3-N-morpholinopropanesulfonic acid, potassium thiocyanate, and dithiothreitol, from a mixture of spores, parasporal crystals, and cell debris of Bacillus thuringiensis subsp . kurstaki . The protein was active against the third instar larvae of Trichoplusia ni, was soluble in 19 mM Na2CO3, and was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed as the insecticidal component of the 132-kDa protoxin of B . thuringiensis subsp . kurstaki by an enzyme-linked immunosorbent assay using antibodies prepared against the protoxin.

Arch Dermatol, 1990 Jun, 126(6), 787 - 90
Bacillary epithelioid angiomatosis occurring in an immunocompetent individual; Cockerell CJ et al.; Within the last several years, a newly characterized condition known as bacillary epithelioid angiomatosis (BEA) has been described in a number of patients with human immunodeficiency virus (HIV) infection . All cases heretofore described have been seen in patients with the HIV infection . We recently evaluated a 37-year-old healthy man who had a localized form of BEA confirmed by biopsy, special strains, electron microscopy, and culture . We conclude that BEA as previously defined may occur in healthy, non-HIV-infected individuals.

J Bacteriol, 1990 Jun, 172(6), 3290 - 7
Mobilization of closely related plasmids pUB110 and pBC16 by Bacillus plasmid pXO503 requires trans-acting open reading frame beta; Selinger LB et al.; Genetic analysis of the closely related nonconjugative plasmids pUB110 and pBC16 has demonstrated that the open reading frame beta (ORF-beta) region in pUB110 and the corresponding homologous region in pBC16 are essential for mobilization of these plasmids by pLS20 or its derivatives . Deletions in this region or insertions that interrupted ORF-beta severely impaired or eliminated the mobilization of pUB110::pUC18 and pBC16::pUC18 hybrids . In contrast, a hybrid in which pUC18 was inserted into pBC16 at a point outside ORF-beta transferred at a frequency comparable to that of intact pUB110 or pBC16 (10(-4) transcipients per donor cell) . The defect of most transfer-deficient (Mob-) hybrid plasmids could be complemented by an intact sister plasmid (i.e., pBC16 for pUB110::pUC18 Mob- hybrids) . The inability to complement certain constructs suggested that the origin of transfer might be located in an area 5' to ORF-beta . Furthermore, cloning the region 5' to ORF-beta onto a nonmobilizable pC194::pUC18 construct resulted in a hybrid plasmid, pUCCoriTBC16, that could be mobilized with complementation . These results indicate that mobilization of pUB110 and pBC16 by conjugative helper plasmids requires ORF-beta in trans and at least one other region, including the RSA sequence, which presumably functions as an origin of transfer, in cis.

J Bacteriol, 1990 Jun, 172(6), 3172 - 9
Identification of beta-exotoxin production, plasmids encoding beta-exotoxin, and a new exotoxin in Bacillus thuringiensis by using high-performance liquid chromatography; Levinson BL et al.; An improved high-performance liquid chromatography separation was developed to detect and quantify beta-exotoxin production in Bacillus thuringiensis culture supernatants . Exotoxin production was assigned to a plasmid in five strains, from three subspecies (B . thuringiensis subsp . thuringiensis serotype 1, B . thuringiensis subsp . tolworthi serotype 9, and B . thuringiensis subsp . darmstadiensis serotype 10) . A new exotoxin, called type II beta-exotoxin in this report, was discovered in B . thuringiensis subsp . morrisoni serotype 8ab, purified, and partially characterized . This material is more specific than type I beta-exotoxin and is very active against the Colorado potato beetle, Leptinotarsa decemlineata.

J Bacteriol, 1990 Jun, 172(6), 2826 - 32
Location of the dipteran specificity region in a lepidopteran-dipteran crystal protein from Bacillus thuringiensis; Widner WR et al.; Two highly related crystal protein genes from Bacillus thuringiensis subsp . kurstaki HD-1, designated cryIIA and cryIIB (previously named cryB1 and cryB2, respectively), were used to study host range specificity . Their respective gene products are 87% identical but exhibit different toxicity spectra; CryIIA is toxic to both mosquito and tobacco hornworm larvae, whereas CryIIB is toxic only to the latter . Hybrids of the cryIIA and cryIIB genes were generated, and their resultant gene products were assayed for toxicity . A short segment of CryIIA corresponding to residues 307 through 382 was shown to be sufficient for altering host range specificity-i.e., when this region replaced the corresponding segment of CryIIB, the resulting hybrid protein acquired toxicity against mosquitoes . The CryIIA and CryIIB polypeptides differ by only 18 amino acids in this region, indicating that very few amino acid changes can have a substantial effect on the toxicity spectra of these proteins.

J Egypt Soc Parasitol, 1990 Jun, 20(1), 197 - 201
Methacrylate embedding as a new technique for histopathological studies on the effect of Bacillus thuringiensis against Culex pipiens; Hussein MA et al.; Studies on the pathogen Bacillus thuringiensis in Egypt and its possible role as a biological control agent have been reviewed . More emphasis in research was given to some lepidopterous cotton pests and few other species . Histopathological changes in B . thuringiensis treated larvae could be observed in the midgut . The methacrylate embedding technique achieved satisfactory results in concern of better preservation of histological fine details.

Appl Environ Microbiol, 1990 Jun, 56(6), 1710 - 6
Molecular cloning of the 130-kilodalton mosquitocidal delta-endotoxin gene of Bacillus thuringiensis subsp . israelensis in Bacillus sphaericus; Trisrisook M et al.; A 3.7-kilobase (kb) XbaI fragment harboring the cryIVB gene (L . Thorne, F . Garduno, T . Thompson, D . Decker, M . A . Zounes, M . Wild, A . M . Walfield, and T . J . Pollock, J . Bacteriol . 166:801-811, 1986) which encoded a 130-kilodalton (kDa) mosquitocidal toxin from a 110-kb plasmid of Bacillus thuringiensis subsp . israelensis 4Q2-72 was cloned into pUC12 and transformed into Escherichia coli . The clone with a recombinant plasmid (designated pBT8) was toxic to Aedes aegypti larvae . The fragment (3.7 kb) was ligated into pBC16 (tetracycline resistant {Tcr}) and transformed by the method of protoplast transformation into Bacillus sphaericus 1593 and 2362, which were highly toxic to Anopheles and Culex mosquito larvae but less toxic to Aedes larvae . After cell regeneration on regeneration medium, the Tcr plasmids from transformants (pBTC1) of both strains of B . sphaericus were prepared and analyzed . The 3.7-kb XbaI fragment from the B . thuringiensis subsp . israelensis plasmid was shown to be present by agarose gel electrophoresis and Southern blot hybridization . In addition, B . sphaericus transformants produced a 130-kDa mosquitocidal toxin which was detected by Western (immuno-) blot analysis with antibody prepared against B . thuringiensis subsp . israelensis 130-kDa mosquitocidal toxin . The 50% lethal concentrations of the transformants of strains 1593 and 2362 against A . aegypti larvae were 2.7 X 10(2) and 5.7 X 10(2) cells per ml, respectively . This level of toxicity was comparable to the 50% lethal concentration of B . thuringiensis subsp . israelensis but much higher than that of B . sphaericus 1593 and 2362 (4.7 X 10(4) cells per ml) against A . aegypti larvae.(ABSTRACT TRUNCATED AT 250 WORDS)

Anal Biochem, 1990 Jun, 187(2), 240 - 5
Determination of phosphatidylcholine in a flow injection system using immobilized enzyme reactors; Masoom M et al.; Two alternative procedures are described for the quantitative determination of phosphatidylcholine in a flow-injection system utilizing immobilized enzymes . Phospholipase C from Bacillus cereus and phospholipase D from cabbage were covalently bound to the surface of controlled-pore glass beads and the enzyme-derivatized beads were packed in small columns . In the first procedure, the phospholipase C column was connected with a second column containing coimmobilized alkaline phosphatase and choline oxidase . In the alternative procedure, the column packed with immobilized phospholipase D was connected with a column packed with immobilized choline oxidase . The hydrogen peroxide produced through the action of choline oxidase in both flow-injection systems was detected amperometrically . Both procedures are suitable for an accurate and rapid quantitation of phosphatidylcholine . The sensitivity of the method based on phospholipase C and alkaline phosphatase is higher than that using phospholipase D . Quantitation of phosphatidylcholine at the nanomole level can be easily obtained using the first method.

Lepr Rev, 1990 Jun, 61(2), 112 - 31
The pathology of the eye in armadillos experimentally infected with Mycobacterium leprae; Brandt F et al.; One hundred and twenty-seven eyes from 66 Mycobacterium leprae inoculated armadillos were studied histologically and some ultrastructurally . Inflammatory reactions were found in the following extraocular tissues: the eyelid, including the orbicularis muscle and the third eyelid, extraocular muscles, tear gland and Harder's gland . The early and slight changes of the intraocular tissues, small amounts of lymphocytes, plasma cells and macrophage infiltrations were confined to the area around the anterior angle specifically within the trabeculae and the adjacent ciliary body, the root of the iris and the limbus region of the cornea . But in the cases with severe lesions the whole uvea was densely infiltrated with large, foamy macrophages intermingled with small amounts of lymphocytes, plasma cells and frequently, neutrophils . No specific necrosis of the granulomas was seen . No explanation for the neutrophil infiltrations was given . The lesions in the cornea were significantly less severe than those in the uvea . Retinal lesions comprised of macrophage infiltrations were all obvious extensions of the adjacent uvea lesions . Acid-fast bacilla (AFB) were found within all tissues . The infection of the intraocular tissues in the armadillo eyes seemed to be mainly, if not solely, haematogenous.

Int J Lepr Other Mycobact Dis, 1990 Jun, 58(2), 273 - 80
Controlled clinical trial of two multidrug regimens with and without rifampin in highly bacilliferous BL/LL south Indian patients: a five-year report; Thomas A et al.; A controlled clinical trial of two multidrug regimens in multibacillary lepromatous and near-lepromatous patients with a bacterial index (BI) of 2.5 or more was conducted . Patients were randomly allocated to either a two-drug regimen of dapsone plus clofazimine for 60 months or a four-drug regimen of rifampin, isoniazid, dapsone, and clofazimine for the first 3 months and clofazimine plus dapsone for the next 57 months . There was no difference between the rifampin and nonrifampin regimens with respect to the clinical improvement or bacteriological status of the patients at 60 months . Reactive states and neuritis were observed to be equal in the two patient groups.

Clin Exp Immunol, 1990 Jun, 80(3), 332 - 8
Strain differences in mouse cellular responses to Mycobacterium lepraemurium and BCG subcutaneous infections . I . Analysis of cell surface phenotype in local granulomas; Roch F et al.; C57BL/6, BALB/c and CBA mice were subcutaneously infected with either Mycobacterium lepraemurium (MLM) or BCG, and studied for bacillary growth, granuloma size of infected footpads and draining lymph nodes (DLN), and DLN cell surface phenotype . Whereas, BCG-infected mice controlled the infection and developed early and large granulomas, MLM-infected mice exhibited major strain variations in their resistance to the infection, as well as in the granuloma size and kinetics . C57BL/6 mice, highly resistant, displayed early and regressive granulomas; BALB/c mice showed lower resistance and early granulomas that grew continuously; CBA mice, highly susceptible, developed late, soft, phagocyte-rich granulomas . Important strain differences in lymph node lymphocyte subset distribution could be observed prior to any infection: C57BL/6 mice displayed higher B cell percentages than both of the other strains and BALB/c mice showed the highest CD4/CD8 ratios, followed by CBA and C57BL/6 mice . BCG and MLM infections both induced similar changes of these parameters in all three strains: that is a decrease of the B cell percentage and a decrease of the CD4/CD8 ratio, and the strain differences observed in uninfected mice persisted . On the other hand, DLN cells stimulated by the infecting bacillus and interleukin 2 also displayed an increase of the CD8 T cell percentage as compared with normal lymph node cells, but this phenomenon was much less pronounced in BALB/c mice, whether infected by MLM or BCG, and in MLM-infected CBA mice, than in BCG- or MLM-infected C57BL/6 (B6) mice . Thus the ability of C57BL/6 mice to generate an early and persistent CD8 T cell response to mycobacteria may contribute to their resistance to MLM.

Mol Gen Mikrobiol Virusol, 1990 Jun, (6), 24 - 8
{Expression of delta-endotoxin gene from Bacillus thuringiensis var . berliner 1715 in Escherichia coli and Bacillus megaterium cells}; Sever IS et al.; Effects of the structure of plasmids carrying the cloned delta-endotoxin gene (tox) ot Bacillus thuringiensis and of the culture media on the expression of the gene have been studied . The DNA region located upstream from the crystal protein gene promoter inhibited the expression of the tox gene in Escherichia coli cells, but enhanced the expression in Bacillus megaterium cells grown in LB medium . The upstream DNA region did not affect the tox gene expression when Bacillus megaterium cells were grown in SSM medium.

FEMS Microbiol Lett, 1990 Jun 1, 57(3), 245 - 8
Inhibition of peptidoglycan biosynthesis in Bacillus megaterium by daptomycin; Mengin-Lecreulx D et al.; The effects of daptomycin on exponential phase cells of Bacillus megaterium were investigated . Bacteriostasis was observed for concentrations between 1 and 3 micrograms/ml and maximal rate of cell lysis at 10 micrograms/ml . At sublytic concentrations (1.5-3 micrograms/ml), the variations of the pools of UDP-N-acetylglucosamine and UDP-N-acetylmuramyl-pentapeptide, as well as the incorporation of (14C)-N-acetylglucosamine into peptidoglycan were studied . From the results it was concluded that the lethal target of daptomycin could be a metabolic step between glucosamine 6-phosphate and UDP-N-acetylglucosamine.

J Clin Microbiol, 1990 Jun, 28(6), 1265 - 70
Restriction endonuclease analysis of Eikenella corrodens; Chen CK et al.; Eikenella corrodens is a gram-negative facultative bacillus commonly found in the oral cavity . Although the role of E . corrodens in periodonititis is not clear, its isolation from extraoral infections attests to its pathogenic potential . Previous studies suggested that this species is phenotypically diverse . In the present study, we used restriction endonuclease analysis (REA) to assess the genetic diversity of this species and to explore the applicability of REA in studying the transmission of E . corrodens . Two groups of E . corrodens isolates were used in this study . Group 1 included 47 epidemiologically independent isolates recovered from dental plaques in periodontally healthy subjects and periodontitis patients and from extraoral infections in different geographic areas . Group 2 E . corrodens included 40 isolates recovered from two periodontitis patients and two periodontally healthy subjects . The results indicated that E . corrodens is genetically heterogeneous, as determined by REA . The majority of the group 1 E . corrodens isolates exhibited strain-specific restriction patterns . Forty restriction patterns were distinguishable among the 47 isolates . Analyses of group 2 isolates revealed that three of four subjects harbored more than one clonal type of E . corrodens . In one instance, a periodontitis patient was found to be colonized by six different clones . Furthermore, two different clonal types of E . corrodens were recovered from a single periodontal pocket in this patient . The results indicated that REA may be a useful tool in the epidemiologic investigation of E . corrodens infections.

Infect Immun, 1990 Jun, 58(6), 1532 - 7
Modification of the glycolipid-binding specificity of vero cytotoxin by polymyxin B and other cyclic amphipathic peptides; Head S et al.; Polymyxin B, an amphipathic cyclic decapeptide produced by Bacillus polymyxa, is routinely used in the extraction of the components from the periplasmic space of gram-negative bacteria . Vero cytotoxin 1 (VT1) is an Escherichia coli-elaborated subunit toxin which binds to the glycolipid globotriosylceramide (Gal-alpha 1-4-Gal beta 1-4-Glc-ceramide {Gb3}) and has been strongly implicated in the etiology of the hemolytic uremic syndrome and hemorrhagic colitis . We now show by in vitro glycolipid-binding assays that in the presence of low concentrations of polymyxin B, globotetraosylceramide (GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc-ceramide {Gb4}) is also recognized by both the VT1 B (binding) subunit and holotoxin . Melittin, a 26-amino-acid cyclic peptide of similar amphipathic nature, produced the same effect, whereas a hydrophobic blocking agent did not . Triton X-100 did not increase binding of VT1 to Gb4 but prevented glycolipid binding in toto at concentrations above 0.5% . Caution is therefore advised in the analysis of VT1 glycolipid binding in the presence of amphipathic peptides.

Rev Esp Enferm Dig, 1990 Jun, 77(6), 409 - 13
{Isolation of M . tuberculosis in abdominal specimens}; Xercavins Valls M et al.; The isolation of mycobacteria in abdominal specimens during a 10 years period is presented . Twenty-three clinical cases have been reviewed; patients were divided in three groups: 1) Peritoneal and intestinal tuberculosis . 2) Pulmonary tuberculosis with isolation of M . tuberculosis in feces, and 3) Miliary tuberculosis . We emphasize the low yielding of bacilloscopy, the low number of colonies in cultures and the importance of the microbiological study of abdominal specimens in the confirmatory diagnosis . The predominant symptoms of peritoneal tuberculosis were abdominal pain and distention and fever . The study of the ascitic fluid showed in most of the cases lymphocytic exudate and the pathological study of biopsies showed granulomas with caseous necrosis . Three patients had another associated abdominal disease . Isolation of M . tuberculosis in feces does not invariably mean the presence of intestinal tuberculosis . We confirm the frequent association of disseminated tuberculosis and HIV1 infection.

An Med Interna, 1990 Jun, 7(6), 312 - 4
{Epithelioid angiomatosis and HIV infection . A neoplastic or infectious proliferative disease?}; Aguilar Ligorit E et al.; A case of "epithelioid angiomatosis" detected in a homosexual patient with positive seric HIV is presented . The skin lesions cytoplasm rich, closed y clumped together, with scanty stromal compound . Warthin-Starry staining {correction of tintions} did not show Bacillus causing "cat scratch disease" . We review and analyse the features of this type of proliferation in patients infected with HIV.

Biomed Environ Sci, 1990 Jun, 3(2), 217 - 39
Chemical management of forest pest epidemics: a case study; Ecobichon DJ; The management of insect epidemics in large tracts of forest is difficult given the climatic conditions encountered, the topography of the forested land, the nature of the forest, the types of chemical and/or biological insecticides registered for use, and the technologies available for insecticide application . Since 1952, the province of New Brunswick, Canada, has been heavily involved in attempting to control an epidemic of the eastern spruce budworm (Choristoneura fumiferana . Clemens) that has ravaged the coniferous softwoods of eastern Canada and the United States . Of the available options, the provincial government chose to develop an aerial spraying program, eventually selecting two chemical insecticides (fenitrothion and aminocarb) and one biological control agent (Bacillus thuringiensis) . Concerns about possible impacts on human health led to extensive studies of the toxicology of these insecticides, the technology of aerial spraying, the development of less hazardous formulations, and the quantitation of off-target drift of aerosolized insecticides . These studies culminated in improvements in pesticide application and the establishment of regulations on safety or buffer zones around human habitation for certain types of aircraft applying different formulations of the insecticides.

J Am Mosq Control Assoc Suppl, 1990 Jun, 2, 1 - 93
The medical importance of riceland mosquitoes and their control using alternatives to chemical insecticides; Lacey LA et al.; The medical importance, ecology and control of riceland mosquitoes using alternative strategies is reviewed . Over 135 pest and vector anopheline and culicine mosquito species found in association with riceland habitats and their medical importance are presented . Malaria and Japanese encephalitis are the two most serious human diseases transmitted by riceland mosquitoes, but they have been incriminated as vectors of dozens of arboviruses and other parasites and pathogens including the causal agents of West Nile and Rift Valley Fevers and lymphatic filariasis . Control of vector and pest mosquitoes using chemical pesticides has generated several problems including: insecticide resistance, safety risks for humans and domestic animals, and other environmental concerns . These problems and the high cost and sustainability of programs based predominantly on conventional insecticides have stimulated increased interest in integrated control measures in ricelands . The integrated pest management (IPM) strategy for mosquito control, also known as integrated vector control (IVC), is an ecologically based approach that may involve several complementary interventions used in combination or singly . Environmental management, and chemical, biological and mechanical control, comprise the elements of IVC proposed for use in or near riceland habitats . Some of the elements of environmental management include the use of intermittent irrigation; flushing of fields; use of rice cultivars that require less water; shifting of planting schedules to avoid optimal mosquito breeding conditions; relocation of communities or use of dry belt farming around them; and zooprophylaxis and other personal protection methods, especially use of insecticide-impregnated bed nets . Biological control agents that have been used successfully in rice fields include several species of larvivorous fish, a mermithid nematode (Romanomermis culicivorax), a fungus (Lagenidium giganteum) and bacteria (Bacillus thuringiensis var . israelensis and Bacillus sphaericus) . The mermithid and the entomopathogens have demonstrated little or no adverse effects on populations of vertebrate and invertebrate nontarget organisms . The successful use of any particular method or combination of interventions for the control of riceland mosquitoes will depend on in-depth ecological studies on the target species and nontarget organisms, sound geographic reconnaissance and effective routine sampling and evaluation . When biological control agents are considered, additional background on the environmental factors limiting their efficacy will also be needed . In addition to the technical components of the various interventions employed in integrated control, sustained suppression of riceland mosquitoes and the diseases they transmit will require a greater sociocultural supportive background, particularly in developing countries.(ABSTRACT TRUNCATED AT 400 WORDS)

J Am Mosq Control Assoc, 1990 Jun, 6(2), 325 - 7
Relative potency of Bacillus thuringiensis var . israelensis and Bacillus sphaericus 2362 for Mansonia titillans and Mansonia dyari; Lord JC et al.; Bacillus thuringiensis var . israelensis (B.t.i.), HD-968-S-1983, and a primary powder of Bacillus sphaericus 2362 were assayed against Mansonia titillans and Ma . dyari . The susceptibility of the 2 species to the individual bacterial toxins was similar . The B.t.i . LC50 values were 50.5 micrograms/liter for Ma . dyari and 54.7 micrograms/liter for Ma . titillans; the B . sphaericus LC50 values were 339.0 micrograms/liter and 347.2 micrograms/liter, respectively . The log dose-probit mortality lines were parallel for both target species . The potency of B . sphaericus relative to B.t.i . was 0.15 for Ma . dyari and 0.16 for Ma . titillans.

Biol Trace Elem Res, 1990 Jun, 25(3), 193 - 9
Mobilization of selenium by a selenium-dependent bacterium; Razak AA et al.; A selenium-dependent bacterium, Bacillus sp., failed to grow on selenium-free media . However, it is able to grow at high concentrations of sodium selenite containing media up to 3% (w/v) . It accumulated extraordinary high quantities of selenium, 432 ppm/mL . The bacterium generated the transformation of inorganic selenium into volatile selenium form(s) into the atmosphere . The biological release of volatile selenium basically depended on several factors: incubation temperature, pH, incubation periods, and substrate concentration . Maximal quantity of the volatile selenium form was obtained at 30 degrees C, pH 7, and 1% (w/v) sodium selenite.

Biol Trace Elem Res, 1990 Jun, 25(3), 187 - 92
Metabolism of selenium in a selenium-dependent bacterium; Razak AA et al.; A selenium-dependent Bacillus sp . is able to grow well up to 3% sodium selenite-containing media . The bacterium completely failed to grow on media devoid of selenium . The presence of selenium in the growth media increased the bacterial contents of proteins, carbohydrates, and lipids . The highest quantities of amino acids were detected at 2% sodium selenite-containing media . The bacterium metabolized selenite into several protein selenoamino acids such as selenomethionine and selenocysteine/selenocystine, as well as nonprotein selenoamino acids, such as selenocystathionine . Several phosphoamino acids were detected in the presence of elevated levels of selenium . The synthesized protein seems not to be affected by the presence of selenium.

FEBS Lett, 1990 May 21, 264(2), 206 - 10
Expression in Escherichia coli of a sub-gene encoding the lipoyl domain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus; Dardel F et al.; A sub-gene encoding the lipoyl domain (residues 1-85) of the lipoate acetyltransferase chain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus was over-expressed in Escherichia coli . Approx . 80% of the domain was unlipoylated but most of the remainder was correctly lipoylated on Lys-42 and could be reductively acetylated by the B stearothermophilus enzyme complex . A small proportion (approx . 4%) of the domain carried an aberrant substituent, possibly an octanoyl group, on Lys-42 . The 400 MHz 1H NMR spectra of the lipoylated and unlipoylated domains were essentially identical and closely resembled that of the native lipoyl domain.

Eur J Biochem, 1990 May 20, 189(3), 523 - 7
Unusual proteolysis of the protoxin and toxin from Bacillus thuringiensis . Structural implications; Choma CT et al.; Trypsin is shown to generate an insecticidal toxin from the 130-kDa protoxin of Bacillus thuringiensis subsp . kurstaki HD-73 by an unusual proteolytic process . Seven specific cleavages are shown to occur in an ordered sequence starting at the C-terminus of the protoxin and proceeding toward the N-terminal region . At each step, C-terminal fragments of approximately 10 kDa are produced and rapidly proteolyzed to small peptides . The sequential proteolysis ends with a 67-kDa toxin which is resistant to further proteolysis . However, the toxin could be specifically split into two fragments by proteinases as it unfolded under denaturing conditions . Papain cleaved the toxin at glycine 327 to give a 34.5-kDa N-terminal fragment and a 32.3-kDa C-terminal fragment . Similar fragments could be generated by elastase and trypsin . The N-terminal fragment corresponds to the conserved N-terminal domain predicted from the gene-deduced sequence analysis of toxins from various subspecies of B . thuringiensis, and the C-terminal fragment is the predicted hypervariable sequence domain . A double-peaked transition was observed for the toxin by differential scanning calorimetry, consistent with two or more independent folding domains . It is concluded that the N- and C-terminal regions of the protoxin are two multidomain regions which give unique structural and biological properties to the molecule.

Arch Biochem Biophys, 1990 May 15, 279(1), 195 - 201
Evidence that chemical modification of a positively charged residue at position 189 causes the loss of catalytic activity of iron-containing and manganese-containing superoxide dismutases; Chan VW et al.; The Escherichia coli, Bacillus stearothermophilus, and human manganese-containing superoxide dismutases (MnSODs) and the E . coli iron-containing superoxide dismutase (FeSOD) are extensively inactivated by treatment with phenylglyoxal, an arginine-specific reagent . Arg-189, the only conserved arginine in the primary sequences of these four enzymes, is also conserved in the three additional FeSODs and five of the six additional MnSODs sequenced to date . The only exception is Saccharomyces cerevisiae MnSOD, in which it is conservatively replaced by lysine . Treatment of S . cerevisiae MnSOD with phenylglyoxal under the same conditions used for the other SODs gives very little inactivation . However, treatment with low levels of 2,4,6-trinitrobenzenesulfonate (TNBS) or acetic anhydride, two lysine-selective reagents that cause a maximum of 60-80% inactivation of the other four SODs, gives complete inactivation of the yeast enzyme . Total inactivation of yeast MnSOD with TNBS correlates with the modification of approximately five lysines per subunit, whereas six to seven acetyl groups per subunit are incorporated on complete inactivation with {14C}-acetic anhydride . It appears that the positive charge contributed by residue 189, lysine in yeast MnSOD and arginine in all other SODs, is critical for the catalytic function of MnSODs and FeSODs.

Experientia, 1990 May 15, 46(5), 475 - 7
The toxicity of two Bacillus thuringiensis delta-endotoxins to gypsy moth larvae is inversely related to the affinity of binding sites on midgut brush border membranes for the toxins; Wolfersberger MG; The delta-endotoxin from Bacillus thuringiensis subspecies kurstaki strain HD1-9 is almost 400 times more potent than the delta-endotoxin from strain HD-73 as a gypsy moth larvicide . The two delta-endotoxins compete for a high-affinity binding site on the brush border membrane of larval gypsy moth midguts . The affinity for the delta-endotoxin from strain HD-73 is much greater than the affinity for the delta-endotoxin from strain HD1-9.

Biochem J, 1990 May 15, 268(1), 15 - 25
Evidence that the inositol phospholipids are necessary for exocytosis . Loss of inositol phospholipids and inhibition of secretion in permeabilized cells caused by a bacterial phospholipase C and removal of ATP; Eberhard DA et al.; We directly manipulated the levels of PtdIns, PtdInsP and PtdInsP2 in digitonin-treated adrenal chromaffin cells with a bacterial phospholipase C (PLC) from Bacillus thuringiensis and by removal of ATP . The PtdIns-PLC acted intracellularly to cause a large decrease in {3H}inositol- or {32P}phosphate-labelled PtdIns, but did not directly hydrolyse PtdInsP or PtdInsP2 . {3H}PtdInsP and {3H}PtdInsP2 levels declined markedly, probably because of the action of phosphatases in the absence of synthesis . Removal of ATP also caused marked decreases in {3H}PtdInsP and {3H}PtdInsP2 . The decrease in polyphosphoinositide levels by PtdIns-PLC treatment or ATP removal was reflected by the inhibition of the production of inositol phosphates upon subsequent activation of the endogenous PLC by Ca2(+)-dependent catecholamine secretion from permeabilized cells was strongly inhibited by PtdIns-PLC treatment and by ATP removal . Ca2(+)-dependent secretion was similarly correlated with the sum of PtdInsP and PtdInsP2 when the level of these lipids was changed by either manipulation . PtdIns-PLC inhibited only the ATP-dependent component of secretion and did not affect ATP-dependent secretion . Both PtdIns-PLC and ATP removal inhibited the late slow phase of secretion, but had little effect on the initial rapid phase . Although we found a tight correlation between polyphosphoinositide levels and secretion, endogenous phospholipase C activity (stimulated by Ca2+, guanine nucleotides and related agents) was not correlated with secretion . Additional experiments indicated that neither the products of the PtdIns-PLC reaction (diacylglycerol and InsP1) nor the inability to generate products by subsequent activation of the endogenous PLC is likely to account for the inhibition of secretion . Incubation of permeabilized cells with neomycin in the absence of ATP maintained the level of polyphosphoinositides and more than doubled subsequent Ca2(+)-dependent secretion . The data suggest that: (1) Ca2(+)-dependent secretion has a requirement for the presence of inositol phospholipids; (2) the enhancement of secretion by ATP results in part from increased polyphosphoinositide levels; and (3) the role for inositol phospholipids in secretion revealed in these experiments is independent of their being substrates for the generation of diacylglycerol and InsP3.

Carbohydr Res, 1990 May 15, 199(1), 83 - 9
Effects of reduced malto-oligosaccharides on the thermal stability of pullulanase from Bacillus acidopullulyticus; Kusano S et al.; We investigated the effects of the reduced malto-oligosaccharides, D-glucitol (G1-OH), maltitol (G2-OH), maltotriitol (G3-OH), maltotetraitol (G4-OH), and maltopentaitol (G5-OH) on the thermal stability of Bacillus acidopullulyticus pullulanase (EC 3.2.1.41) . The thermal stability depended on the concentration of D-glucitol; after heat treatment for 90 min at 60 degrees in the presence of 0.56, 0.28, 0.14, or 0M G1-OH, the residual activity was 100, 80, 32, and 10% of the control, respectively . Stability increased with the number of glucosyl residues in the alditols added; the effects of G3-OH, G4-OH, and G5-OH on stability were remarkable . Addition of 30% G2-OH, G3-OH, and G4-OH also contributed to the thermal stability of the pullulanase immobilized onto chitosan beads treated with glutaraldehyde . A high concentration of G2-OH stabilized other debranching amylases, Klebsiella pneumoniae pullulanse, Bacillus sectorramus pullulanase, and Pseudomonas amyloderamosa isoamylase (EC 3.2.1.68) under heat treatment for 48 h at 60 degrees, as well as the pullullanase of B . acidopullulyticus.

J Appl Bacteriol, 1990 May, 68(5), 461 - 9
The effects of controlled exposure to formaldehyde vapour on spores of Bacillus globigii NCTC 10073; Cross GL et al.; The effects of exposure of spores of Bacillus globigii NCTC 10073 to controlled levels of formaldehyde vapour under varying environmental conditions are reported . The death rate of the micro-organism varies with the humidity and formaldehyde concentration and can be predicted for conditions within the limits of the information available . A mathematical model for the effects of formaldehyde is proposed which can be used to calculate the effects of known conditions and as the basis of a controlled formaldehyde fumigation process . Results of practical tests of such a process are described.

J Protozool, 1990 May-Jun, 37(3), 168 - 73
The interaction of Naegleria fowleri amoebae with murine macrophage cell lines; Fischer-Stenger K et al.; The present study was undertaken to determine whether murine macrophage cell lines exhibited in vitro amoebicidal activity comparable to that elicited by activated murine peritoneal macrophages . Peritoneal macrophages activated in vivo by bacillus Calmette-Guerin or Propionibacterium acnes demonstrated significant cytolysis of Naegleria fowleri amoebae . The macrophage cell line RAW264.7 also effected cytolysis of amoebae, but to a lesser extent than that elicited by activated peritoneal macrophages . However, the macrophage cell lines, J774A.1 and P388D1, did not exhibit amoebicidal activity . Macrophage conditioned medium prepared from RAW264.7 macrophages mediated cytolysis of L929 tumor cells but had no effect on N . fowleri amoebae . In addition, neither recombinant tumor necrosis factor nor recombinant interleukin-1 exhibited amoebicidal activity . Scanning electron microscopy of co-cultures revealed that N . fowleri bound to activated peritoneal macrophages and RAW264.7 macrophages . These results suggest that RAW264.7 macrophages treated in vitro with lipopolysaccharide are similar to macrophages activated in vivo in that they effect contact-dependent cytolysis of Naegleria fowleri amoebae . The RAW264.7 macrophages are unlike primary macrophage cultures in that they either do not release soluble amoebicidal factors into the conditioned medium or they release insufficient quantities.

J Invertebr Pathol, 1990 May, 55(3), 387 - 93
Bacillus thuringiensis potency bioassays against Heliothis armigera, Earias insulana, and Spodoptera littoralis larvae based on standardized diets; Navon A et al.; Bacillus thuringiensis screening programs based on the official potency bioassay using third-instar larvae and on a neonate bioassay were developed for Heliothis armigera, Earias insulana, and Spondoptera littoralis . In these bioassays, the diets were standardized to be suitable, with minor modifications, for feeding of the three lepidopterans . The bioassay protocol was based on determination of the LC50 of the microbial standard HD-1-S-80 in the insects susceptible to B . thuringiensis var . kurstaki strains . This was followed by preliminary screening of B . thuringiensis strains at the LC50 of the B . thuringiensis standard . The B . thuringiensis strains causing 100% mortality at this LC50 in the larvae were selected for potency determinations . The neonate bioassay was suitable for accurate determinations of potencies also in S . littoralis--a representative of insects weakly susceptible to the HD-1 standard . The role of the official and the neonate bioassays in developing microbial control programs is discussed.

Appl Environ Microbiol, 1990 May, 56(5), 1378 - 85
Receptors on the brush border membrane of the insect midgut as determinants of the specificity of Bacillus thuringiensis delta-endotoxins; Van Rie J et al.; To investigate the biochemical basis of the differences in the insecticidal spectrum of Bacillus thuringiensis insecticidal crystal proteins (ICPs), we performed membrane binding and toxicity assays with three different ICPs and three lepidopteran species . The three ICPs have different toxicity patterns in the three selected target species . Binding studies with these 125I-labeled ICPs revealed high-affinity saturable binding to brush border membrane vesicles of the sensitive species . ICPs with no toxicity against a given species did not bind saturably to vesicles of that species . Together with previous data that showed a correlation between toxicity and ICP binding, our data support the statement that differences in midgut ICP receptors are a major determinant of differences in the insecticidal spectrum of the entire lepidopteran-specific ICP family . Receptor site heterogeneity in the insect midgut occurs frequently and results in sensitivity to more than one type of ICP.

J Infect Dis, 1990 May, 161(5), 988 - 91
Serum tumor necrosis factor levels and disease dissemination in leprosy and leishmaniasis; Pisa P et al.; It has been suggested that tumor necrosis factor alpha (TNF alpha) may serve as an important antigen-independent host defense mechanism against parasitic organisms . Sera from 66 patients with leishmaniasis and 68 patients with leprosy, all from Ethiopia, were tested for TNF alpha using an enzyme-linked immunoassay . Sera from patients with the multi-parasitic/bacillary type of disease (visceral or diffuse cutaneous leishmaniasis and lepromatous leprosy), known to be associated with absent or low specific T cell response, contained significantly higher TNF alpha titers than those of patients with pauci-parasitic/bacillary disease (localized cutaneous leishmaniasis and nonlepromatous leprosy) . High titers of TNF alpha in the absence of a functioning T cell response do not appear to confer resistance against Leishmania aethiopica and Mycobacterium leprae.

Mikrobiologiia, 1990 May-Jun, 59(3), 453 - 9
{Growth and spore germination factors of various strains of Bacillus sphaericus}; Shevtsov VV et al.; The work was aimed at studying the requirements of sixteen Bacillus sphaericus strains with a different larvicidal activity in amino acids and some other compounds necessary for their growth and spore germination . Most of the strains were found to require arginine, glutamate, methionine, threonine, serine, glycine, alanine and lysine, but they did not assimilate phenylalanine and proline . Arginine, methionine and glutamate were shown to be the most effective inductors of spore germination . Specific differences were detected in the requirements of virulent and avirulent strains . Glucose repressed both spore germination and spore formation.

Mikrobiol Zh, 1990 May-Jun, 52(3), 41 - 4
{The characteristics of the biosynthesis of surface-active lipids by a Bacillus sp . culture}; Eliseev SA et al.; It is established that the Bacillus sp . C-14 strain is able to produce the biosurfactant of lipopeptide nature which decreases the surface and interfacial tension to 29.5 and 2.0 mH/m, respectively . The biosurfactant synthesis is shown to depend on the substrate nature, N:C ratio in the medium as well as on the presence of sodium citrate as a stimulator of lipogenesis.

Mikrobiologiia, 1990 May-Jun, 59(3), 448 - 52
{Correlation between insecticidal and antibiotic activities of Bacillus thuringiensis parasporal crystals}; Egorov NS et al.; The work was concerned with studying the insecticide and antibiotic activities of solutions containing parasporal crystals of several Bacillus thuringiensis subspecies . The correlation coefficients of these values were determined . A direct correlation was established between the insecticide and antibiotic activities of solutions of parasporal crystals from several B . thuringiensis subspecies . Therefore, the quality of bacterial insecticides can be controlled by assaying the antibiotic activity of polypeptide crystals in biotechnology.

J Invertebr Pathol, 1990 May, 55(3), 350 - 6
Studies on the cellular defense reactions of the madeira cockroach, Leucophaea maderae: in vitro phagocytosis of different strains of Bacillus cereus and their effect on hemocyte viability; Rahmet-Alla M et al.; Monolayers of Leucophaea maderae hemocytes, consisting of mainly plasmatocytes and coagulocytes, were incubated with three strains of Bacillus cereus of differing pathogenicities, and the levels of phagocytosis and hemocyte viability were determined . Incubation with viable B . cereus strains NCTC 2599, NCIB 3329, and B1 resulted in a significant drop in hemocyte viability after 60 min of incubation compared with the saline-only controls . The greatest effect, however, resulted from incubation with B . cereus B1 which is the most pathogenic of the three strains studied . The killing effect of the three B . cereus strains was abolished following their UV irradiation . Incubation of monolayers with viable B . cereus B1 resulted in a level of phagocytic activity at all time periods lower than that with the other two strains . The highest levels of phagocytosis were achieved with UV-killed B . cereus, although no significant differences were found in these values between the three strains at any of the incubation times . Phospholipase C, a lytic enzyme shown to be released by all three strains of B . cereus, although in varying amounts, also caused hemocyte death following its incubation with the monolayers . These data suggest that the hemocyte killing and resistance to phagocytosis by B . cereus are caused by the release of phospholipase C, or some other related toxin(s) by the bacteria.

J Bacteriol, 1990 May, 172(5), 2584 - 93
Sequencing the gene for an imipenem-cefoxitin-hydrolyzing enzyme (CfiA) from Bacteroides fragilis TAL2480 reveals strong similarity between CfiA and Bacillus cereus beta-lactamase II; Thompson JS et al.; Using a newly constructed Bacteroides fragilis-Escherichia coli cloning shuttle vector, pJST61, we have cloned the cefoxitin (FOX)-imipenem (IMP) resistance determinant from B . fragilis TAL2480 . FOX-IMP resistance in this strain results from the production of a periplasmic, Zn2(+)-containing beta-lactamase which hydrolyzes carbapenems and cephamycins and whose activity is resistant to clavulanic acid but sensitive to Zn2(+)-binding reagents, including EDTA . The pJST61 vector permits efficient library construction in E . coli and allows for the transfer of the library to B . fragilis recipients for the screening or selection of specific phenotypes . The library clone containing the FOX-IMP resistance gene was detected after transfer to B . fragilis TM4000 (Fox-Imps) selecting for Foxr . One of the isolates carrying plasmid pJST241 is resistant to FOX and IMP and synthesizes a periplasmic protein with substrate and inhibitor properties identical to those of strain TAL2480 . On the basis of deletion analysis, Tn1000 insertion mutations, and DNA sequencing, we have defined the 747-base cfiA (FOX-IMP resistance) gene within the 3.6-kilobase cloned insert in pJST241 . The cfiA gene contains an open reading frame that could code for a precursor protein of 249 amino acids and with a molecular mass of 27,260 daltons . A potential signal sequence has been identified at the N terminus of this protein; cleavage within this sequence would result in a protein of 231 amino acids with a molecular mass of 25,249 daltons . The CfiA protein shows remarkable similarities to the exported, Zn2(+)-requiring, type II beta-lactamase Blm proteins from Bacillus cereus 569/H and 5/B/6 . Although overall amino acid identity is only 32%, the Zn ligand-binding His and Cys residues are precisely conserved and the amino acids in the vicinity of these sites show strong similarities (greater than 80%) when the CfiA and Blm proteins are compared.

Acta Gastroenterol Belg, 1990 May-Jun, 53(3), 307 - 14
{Tuberculous peritonitis . Description of 2 cases associated with another infection and literature review}; Arend P et al.; The authors report two cases of peritoneal tuberculosis observed in Belgium: a 28-year-old black man, immigrant from Ghana and a 24-year-old caucasian woman . In both patients, tuberculosis was associated with another infection . The first patient had a retroperitoneal amoebic cyst and in the second patient Chlamydia trachomatis was found with Koch bacillus in the ascitic fluid . Both patients were successfully treated with appropriate antibiotics . Following the review of recent literature, it appears that the incidence of extrapulmonary tuberculosis is increasing in the industrialized part of the world, peritoneal tuberculosis now representing 0.5% of all cases of tuberculosis . Mortality approaches 7% . The diagnosis is difficult and should be evoked more often in order to be confirmed by appropriate means, such as ascitic fluid adenosine desaminase, bacteriology and peritoneoscopy.

Indian J Med Res, 1990 May, 91, 223 - 7
Characterization & larvicidal activity of indigenous isolates of Bacillus sphaericus from natural breeding habitats; Manonmani AM et al.; Soil, water and moribund/or dead mosquito larval samples collected from various mosquito breeding habitats of Pondicherry and Tamil Nadu were screened for the presence of mosquito pathogenic B . sphaericus isolates . From 1892 samples 21 isolates were obtained . All these isolates fell under 3 serogroups viz., H5a5b, H6 and H45, the latter two serotypes not reported hitherto as toxic to mosquito larvae and three phage-groups namely, phage group 2, phage group 3 and an unknown one . Twelve of the isolates were highly toxic and superior to the standard strains 1593, 2297 and 2362 supplied by Pasteur Institute, Paris when tested against Culex quinquefasciatus larvae . Out of these 12 strains 11 belonged to the serotype H5a5b and the phage-group 3 and 1 belonged to the serotype H45 and an unknown phage group.

Acta Trop, 1990 May, 47(4), 189 - 95
Evaluation of Bactimos wettable powder, granules and briquets against mosquito larvae in Malaysia; Sulaiman S et al.; There was high mortality in late larval instars of Aedes albopictus (Skuse) from laboratory and field populations in the 24 h after application of three Bactimos formulations of Bacillus thuringiensis H-14 . Mortalities were higher and residual effects longer in field populations than in laboratory ones . Briquets were the most effective formulation (mortality 96-100% after five weeks; 76-92% after eight weeks) . Culex quinquefasciatus Say larvae were tested only against the briquet formulation . In the laboratory, 100% mortality of late instars persisted for six weeks and dropped to 48-88% after eight weeks . In the field, late instars were reduced by 62-87% after 24 h and 69-72% after one week compared to increases in an untreated population of 160% and 176% respectively.

Am J Pathol, 1990 May, 136(5), 1125 - 35
Bartonella bacilliformis stimulates endothelial cells in vitro and is angiogenic in vivo; Garcia FU et al.; Bartonellosis, a biphasic disease caused by motile intracellular bacteria, produces in its tissue phase a characteristic dermal eruption (Verruga peruana) resulting from a pronounced endothelial cell proliferation . Bacteria are found in the interstitium and within the cytoplasm of endothelial cells (Rocha-Lima inclusion) . The aim of this study was to determine if Bartonella bacilliformis produce a substance(s) that might be responsible for the vascular proliferation seen in the Verruga . This was assessed in an in vitro system using human endothelial cells and measuring proliferation as well as production of tissue type plasminogen activator after exposure to the endothelial cultures to B . bacilliformis extracts . Our results indicate that B . bacilliformis possess an activity that stimulates endothelial cell proliferation up to three times that of control . The factor(s) is specific for endothelial cells, heat sensitive, larger than 12 to 14 kd, not enhanced by heparin, has no affinity for heparin, and is precipitated by 45% ammonium sulfate . In addition, the B . bacilliformis extracts stimulate production of t-PA antigen in a concentration-dependent fashion . This activity is also heat sensitive and not lost after dialysis (12 to 14 kd) . B . bacilliformis extracts, however, do not increase the production of plasminogen activator inhibitor . It was also determined that B . bacilliformis extracts stimulate the formation of new blood vessels in an in vivo model for angiogenesis . These results describe a bacterial factor(s) that stimulates two important steps in the development of new blood vessels in vitro, as well as the formation of new blood vessels in vivo . Determining the mechanism of action, combined with a complete characterization of this factor(s), may help in understanding the pathogenesis not only of the Verruga and angiogenesis in general but also the recently described Cat-Scratch-associated epithelioid hemangiomas in patients with AIDS and Kaposi sarcoma.

Hum Pathol, 1990 May, 21(5), 567 - 9
Bacillary angiomatosis presenting as a soft-tissue tumor without skin involvement; Schinella RA et al.; A patient with human immunodeficiency virus infection presented with a soft-tissue mass which histologically and clinically mimicked an angiosarcoma . Ultrastructural study, however, revealed bacteria identical to those seen in cutaneous bacillary angiomatosis, but the patient had no skin lesions . To our knowledge, this represents the first report of soft tissue involvement by bacillary angiomatosis without the presence of skin lesions.

Ugeskr Laeger, 1990 Apr 23, 152(17), 1226 - 8
{Tuberculosis in children in Copenhagen during 1975-1985}; Mortensen J et al.; During the years 1975-1985, 25 Danish children (less than 15 years) and 40 children of immigrants in the Municipalities of Copenhagen and Frederiksberg had tuberculosis for the first time . The children were assessed at the time of diagnosis and were retraced in 1987 . The incidence of tuberculosis among foreign children was 10-40 times that among Danish children . Pronounced differences in the localizations of tuberculosis in Danes and foreigners were observed . Whereas the Danish children had exclusively pulmonary tuberculosis, 13 of the foreign children had either extrapulmonary tuberculosis or pulmonary and extrapulmonary tuberculosis simultaneously . Two of the foreign children had tuberculosis meningitis and one miliary tuberculosis . Significantly more patients with bacillary tuberculosis were present among foreigners than among Danes . No significant differences were found between the occurrence of symptoms, Mantoux-positive reactions, previous Calmette vaccination or known contact with a person with tuberculosis in Danes and foreigners . Treatment was followed as prescribed and all were cured from tuberculosis . One child with tuberculosis meningitis developed slight sequelae while the remainder of the children developed somatically normally.

Biochem J, 1990 Apr 15, 267(2), 309 - 15
Characterization of the cysteine residues and disulphide linkages in the protein crystal of Bacillus thuringiensis; Bietlot HP et al.; Bacillus thuringiensis produces a 130-140 kDa insecticidal protein in the form of a bipyramidal crystal . The protein in the crystals from the subspecies kurstaki HD-1 and entomocidus was found to contain 16-18 cysteine residues per molecule, present primarily in the disulphide form as cystine . Evidence that all the cysteine residues form symmetrical interchain disulphide linkages in the protein crystal was obtained from the following results: (i) the disulphide diagonal procedure {Brown & Hartley (1966) Biochem . J . 101, 214-228} gave only unpaired cysteic acid peptides in diagonal maps; (ii) the disulphide bridges were shown to be labile in dilute alkali and the crystal protein could be released quantitatively with 1 mM-2-mercaptoethanol; (iii) the thiol groups of the released crystal protein were shown by competitive labelling {Kaplan, Stevenson & Hartley (1971) Biochem . J . 124, 289-299} to have the same chemical properties as exposed groups on the surface of the protein; (iv) the thiol groups in the released crystal protein reacted quantitatively with iodoacetate or iodoacetamide . The finding that all the disulphide linkages in the protein crystal are interchain and symmetrical accounts for its alkali-lability and for the high degree of conservation in the primary structure of the cystine-containing regions of the protein from various subspecies.

J Ind Microbiol, 1990 Apr-May, 5(2-3), 95 - 101
Properties of the biosurfactant produced by Bacillus licheniformis strain JF-2; McInerney MJ et al.; The activity of the biosurfactant produced by Bacillus licheniformis strain JF-2 was quantified using a unit defined as the amount of the acid-precipitated biosurfactant that lowered the surface tension by 10 mN/m . One unit was equivalent to 37 micrograms/ml of the acid-precipitated biosurfactant . Acid precipitation was very effective in the removal of the biosurfactant from the spent medium . Among the solvents tested methanol was the most efficient in extracting the surfactant activity from acid-precipitated material . Thin-layer chromatography of the acid-precipitated biosurfactant revealed four components, two of which contained a lipid moiety and one of which contained an amino group . The methanol-soluble fraction also contained these four components . Studies suggested that all four components were needed for full activity . The lowest interfacial tensions against octane were observed when NaC1 concentrations were 50 g/l or greater . Calcium concentrations greater than 25 g/l significantly increased the interfacial tension.

Biochim Biophys Acta, 1990 Apr 6, 1048(2-3), 223 - 30
Cloning and expression of raw-starch-digesting alpha-amylase gene from Bacillus circulans F-2 in Escherichia coli; Kim CH et al.; The raw potato-starch-digesting alpha-amylase gene of Bacillus circulans F-2 was cloned for the first time in Escherichia coli C600, using plasmid pYEJ001 . The recombinant plasmid, named pYKA3, has a 5.4 kb insert from a chromosome of the donor bacterium . Subcloning of this amylase gene gave plasmid pHA300 which carried 3.15 kb of the inserted DNA . The transformed bacterium, E . coli C600 (pYKA3), produced the amylase in the periplasmic space, whereas it is secreted outside the cell in the donor bacterium . The cloned raw-starch-digesting alpha-amylase has a molecular weight of 93,000 on SDS-PAGE, and its action pattern was absolutely the same as that of the potent raw-starch-digestible amylase produced by B . circulans F-2 . The periplasmic amylase produced by the transformed E . coli (pHA300) could digest raw starch granules such as potato, corn and barley raw starch granules, indicating that the raw-starch-digesting amylase is active in E . coli . Furthermore, this amylase crossreacted with the rabbit antiserum raised against the raw potato-digesting alpha-amylase of B . circulans F-2 . From these results it was concluded that the cloned amylase is the same amylase protein as B . circulans F-2 amylase, which has a potent raw-starch digestibility . Thus, this paper is to our knowledge the first describing the molecular cloning of raw-starch-digesting alpha-amylase from Bacillus species and its successful expression in E . coli.

Wei Sheng Wu Xue Bao, 1990 Apr, 30(2), 158 - 60
{Presence of flagellar antigenic subfactors in serotype 1 of Bacillus thuringiensis}; Wang Y et al.; The strain G was isolated from a dead army worm (Leucania separata Walker) in corn field, at Hebei . It has the typical morphological characteristics of Bacillus thuringiensis . The serological analysis showed that the flagellar suspensions of the strain G was agglutinated only by the antisera against the H1 antigen and not by the antisera against the other 22 known H-antigens . The agglutination titre was 1:6,400 . The flagellar suspension of the serotype 1, subsp . thuringiensis, was agglutinated by the antiserum against the strain G, with a titre of 1:12,800 . However, the cross-saturation of antisera showed that there were different fractions in the H1 antigen . The H antiserum against subsp . thuringiensis still agglutinated with subsp . thuringiensis, having a titre of 1:6,400, even if it was fully saturated by strain G . When it was saturated by subsp . thuringiensis the H antiserum of strain G did not agglutinate by himself . Therefore, the H1 antigen (subsp . thuringiensis) must be divided into subfactors H1alb and strain G antigen is H1a . On the basis of this study, we propose the strain G represents the serotype H1a under the name of Bacillus thuringiensis subsp . hebeiensis.

J Biochem (Tokyo), 1990 Apr, 107(4), 645 - 9
Cloning and nucleotide sequences of the BanI restriction-modification genes in Bacillus aneurinolyticus; Maekawa Y et al.; The genes of the BanI restriction-modification system specific for GGPyPuCC were cloned from the chromosomal DNA of Bacillus aneurinolyticus IAM1077, and the coding regions were assigned on the nucleotide sequence on the basis of the N-terminal amino acid sequences and molecular weights of the enzymes . The restriction and modification genes coded for polypeptides with calculated molecular weights of 39,841 and 42,637, respectively . Both the enzymes were coded by the same DNA strand . The restriction gene was located upstream of the methylase gene, separated by 21 bp . The cloned genes were significantly expressed in E . coli cells, so that the respective enzymes could be purified to homogeneity . Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration indicated that the catalytically active form of the endonuclease was dimeric and that of the methylase was monomeric . Comparison of the amino acid sequences revealed no significant homology between the endonuclease and methylase, though both enzymes recognize the same target sequence . Sequence comparison with other related enzymes indicated that BanI methylase contains sequences common to cytosine-specific methylases.

Vet Microbiol, 1990 Apr, 22(2-3), 291 - 7
Cultivation of Bacillus piliformis (Tyzzer) in mouse fibroblasts (3T3 cells); Spencer TH et al.; Bacillus piliformis was successfully propagated in a continuous mouse-embryo fibroblast cell line (3T3) . 3T3 monolayers were successfully inoculated with a variety of infected materials including yolk sac and liver suspensions, minced yolk sac and liver, and primary chicken-embryo liver-cell cultures . An initial decrease in B . piliformis number was noted after inoculation of the cell layer with 2.6 X 10(5) organisms followed by a peak bacterial count at 48 h . Bacillus piliformis remained infectious for mice after 22 passages in 3T3 cell monolayers . Limited growth of B . piliformis was obtained in cell lines of mouse connective-tissue origin (L-929) and mouse liver origin (NCTC 1469).

Vet Microbiol, 1990 Apr, 22(2-3), 237 - 40
Inhibition of Dermatophilus congolensis by substances produced by bacteria found on the skin; Kingali JM et al.; Bacteria, isolated from the skins of clinically normal sheep, were tested for inhibitory activity against Dermatophilus congolensis grown in vitro . Out of 85 bacterial isolates, 19, mainly Bacillus spp., showed zones of inhibition when grown together with D . congolensis . The inhibitory activity was shown to be due to the metabolites released by the bacteria.

Planta Med, 1990 Apr, 56(2), 152 - 7
Effects of Phaeodactylum tricornutum and Dunaliella tertiolecta extracts on the central nervous system; Laguna MR et al.; We report the effects of aqueous extracts of the microalgae Dunaliella tertiolecta Butcher (Chlorophyceae) and Phaeodactylum tricornutum Bohlin (Bacillariophyceae) on oxotremorin-induced cholinergic symptoms, amphetamine-induced hypermotility, pentylenetetrazole-induced convulsions, and Rota-rod test performance . Both extracts significantly reduced rectal temperature, spontaneous motor activity, and time on the Rota-rod and increased the number of mice falling off the rod before 180 s, but neither protected against pentylenetetrazole-induced convulsions or against oxotremorin-induced tremor, salivation, and diarrhoea.

J Econ Entomol, 1990 Apr, 83(2), 347 - 55
Clearance of Bacillus sphaericus and Bacillus thuringiensis ssp . israelensis from mammals; Siegel JP et al.; The maximum recovery period following topical ocular instillation and intraperitoneal injection of two preparations of Bacillus thuringiensis ssp . israelensis de Barjac and two preparations of Bacillus sphaericus 2362 was evaluated in rabbits and mice . B . sphaericus 2362 persisted for 8 wk after administration to the conjunctival cul-de-sac of rabbits; B . thuringiensis ssp . israelensis persisted for 1 wk . Infection was not evident, but both entomopathogens were recovered from flushed and unflushed eyes . High doses of B . sphaericus 2362 (greater than or equal to 10(8) colony-forming units) were toxic to CD-1 mice, and the toxic factor was heat stable . Injection of 10(7) colony-forming units of B . sphaericus 2362 resulted in clearance from the spleens of euthymic and athymic mice . Recovery occurred up to 67 d after injection . Mice failed to remove one preparation of B . thuringiensis ssp . israelensis from their spleen, and a constant number of colony-forming units were recovered for 80 d . B . sphaericus 2362 and B . thuringiensis ssp . israelensis were recovered from heart blood; their disappearance from heart blood coincided with their clearance from the spleen . There was no evidence that either organism was infectious . We conclude that these organisms can be used safely in environments where human exposure might occur.

Am Rev Respir Dis, 1990 Apr, 141(4 Pt 1), 815 - 20
Choosing an appropriate criterion for true or false conversion in serial tuberculin testing; De March-Ayuela P; This study evaluates five different criteria of true conversion in relation to the annual tuberculosis infection rate or risk of infection . It also examines prospective results of a general population, 16 to 69 yr of age, who had less than 6 mm of induration at first PPD test . This population received a second PPD test between 1976 and 1986 . In about 95% of cases, the interval was 1 to 5 yr . In 1,355 bacillus Calmette-Guerin (BCG)-vaccinated subjects, none of the criteria is acceptable for true conversion . Among 1,834 nonvaccinated BCG, only the criterion of a change from less than 10 to 18 mm or more with an increase of 12 mm or more, or the criterion of change from less than 10 to 18 mm or more with an increase of 18 mm or more, is compatible with true conversion and with the correspondent annual risk of infection . But we must accept that with smaller increases, recently infected persons may escape diagnosis of infection . A third test was performed in 81 subjects with two negative test results . In all cases except one, the annual conversion rate observed was much higher than the annual risk of infection . These data reveal shortcomings of the current criterion for conversion . They show that the tuberculin conversion at retesting is not epidemiologically related to new infection and that none of the criteria assures the discovery of true converters . Other factors must be taken into account.

Am J Pathol, 1990 Apr, 136(4), 949 - 56
Effects of in vivo 'priming' on endotoxin-induced hypotension and tissue injury . The role of PAF and tumor necrosis factor; Sun XM et al.; Exogenously administered tumor necrosis factor-alpha (TNF) and bacterial endotoxin (LPS) induce shock and tissue injury . Here, the authors studied the effect of endogenous TNF on LPS-induced hypotension and tissue injury and investigated the role of PAF in these responses . Rats were primed with intraperitoneal injection of zymosan 24 hours before, or Bacillus Calmette-Guerin (BCG) 12 to 15 days before intravenous injection of low dose (0.5 mg/kg) LPS . It was found that nonprimed animals showed mild hypotension and moderate leukopenia in response to LPS . In contrast, zymosanprimed rats developed shock and marked leukopenia, and more severe bowel injury than nonprimed rats . The authors then showed that, following LPS injection, zymosan-primed animals had higher TNF and platelet-activating factor (PAF) levels than nonprimed rats . Pretreatment of the animal with PAF antagonist, SRI 63-441, markedly ameliorated the hypotension and tissue injury . Interestingly, BCG-primed rats did not show aggravation of LPS-induced hypotension . Only TNF (but not PAF) level in these animals was increased . Thus, it appears that TNF release alone, without a sufficient increase in PAF, is incapable of causing severe hypotension . However, most of the BCG-primed animals showed tissue injury, which could be prevented by pretreatment with PAF antagonist . The authors discuss the possible mechanisms of this discrepancy between systemic and local responses in BCG-primed animals.

J Bacteriol, 1990 Apr, 172(4), 2168 - 71
Parasporal bodies of Bacillus laterosporus sporangia; Montaldi FA et al.; Intact colonies of Bacillus laterosporus examined by thin-section transmission electron microscopy revealed sporangia in various stages of development and degeneration as the endospores matured . The sporangia formed a surface layer of hexagonally arranged subunits . The variety of parasporal bodies raised questions of developmental and ecologic utility.

Infect Immun, 1990 Apr, 58(4), 1010 - 6
Protein and antigenic heterogeneity among isolates of Bacillus piliformis; Riley LK et al.; Protein and antigenic heterogeneity among isolates of Bacillus piliformis, the etiologic agent of Tyzzer's disease, were investigated . The seven isolates utilized in this study were originally isolated from naturally infected animals of different animal species and diverse geographical locations . Isolates were propagated in mammalian cell lines, and bacterial extracts were prepared . Protein and antigenic profiles were compared among isolates, using Coomassie blue-stained polyacrylamide gels and Western blot (immunoblot) analyses, respectively . Results showed differences in protein and antigen banding patterns, indicating diversity among isolates . Western blots probed with serum preabsorbed with a heterologous bacterial extract revealed that numerous antigens have different electrophoretic mobilities among isolates but apparently share common epitopes . Immunodominant cross-reactive antigens may be candidate proteins useful for development of improved serologic diagnostic tests, allowing identification of animals infected with a wide range of B . piliformis isolates.

J Urol, 1990 Apr, 143(4), 710 - 2; discussion 712-3
The response of patients with superficial bladder cancer to a second course of intravesical bacillus Calmette-Guerin; Bretton PR et al.; Of 347 patients who received an initial 6-week course of intravesical bacillus Calmette-Guerin for superficial transitional cell carcinoma of the bladder 28 (8%) were treated with another course . Subsequent progression of disease (muscle infiltration, metastasis or local progression) occurred in 13 patients (46%) . Of the 15 patients (54%) without progression 10 (36%) had a complete response and 5 (33%) had new tumors, and they were rendered free of disease after transurethral resection . The median duration of response to course 1 of bacillus Calmette-Guerin was shorter for patients with disease progression after course 2 than for those with no progression (15 and 27 months, respectively, p equals 0.05) . The median followup after course 2 was 31.2 months (range 15.6 to 56.4 months) . The median interval between courses 1 and 2 of intravesical bacillus Calmette-Guerin was 21.6 months (range 3.6 to 78.8 months) . The interval from course 2 of bacillus Calmette-Guerin to progression correlated with the duration of response to course 1 of treatment (p equals 0.01) . It appears that a subsequent treatment with bacillus Calmette-Guerin is most likely to be useful in patients who have a sustained response to the initial treatment.

Pathology, 1990 Apr, 22(2), 106 - 7
Mycobacterium terrae tenosynovitis; Karthigasu KT et al.; Atypical mycobacteria causing extra-pulmonary disease in man are well documented . These infections are manifested by the presence of ulcers, abscesses and lymphadenitis . Mycobacterium marinum is particularly noted for infections involving the synovium, tendon sheaths, bursae and bone . Of lesser note is Mycobacterium terrae (radish bacillus), a nonchromogen also associated with tenosynovitis . We are not aware of any previous report of the association of M . terrae with synovitis in Australia . This case report describes a culture-proven case of tenosynovitis caused by M . terrae.

Int J Syst Bacteriol, 1990 Apr, 40(2), 123 - 5
Description of Bacillus naganoensis sp . nov; Tomimura E et al.; A new species, Bacillus naganoensis, is proposed for an obligately aerobic, moderately acidophilic, endospore-forming bacterium that produces a thermostable, aciduric pullulanase (EC 3.2.1.41) . The organism was isolated from soil by selection on solid, pullulan-containing medium at pH 4.0 and 30 degrees C . The isolate required a medium pH of less than 6.5 for growth initiation . Fatty acid composition studies revealed that the major fatty acid of cells grown in nutrient broth supplemented with 1% starch was 14-methylpentadecanoic acid (iso-C16) at 45 mol% . The guanine-plus-cytosine content of the DNA of this organism was 45 +/- 2 mol% . A type culture has been deposited with the American Type Culture Collection, Rockville, Md., as strain ATCC 53909.

Indian J Lepr, 1990 Apr-Jun, 62(2), 169 - 79
Cultivation in vitro of acid-fast nocardioform chemoautotrophic bacteria from mouse foot-pads infected with human strains of leprosy bacillus; Chakrabarty AN et al.; Four acid-fast nocardioform bacteria could be isolated and cultivated as pure cultures in vitro from mouse foot-pads (MFP), which were infected with serially passaged strains of human leprosy bacillus; the liquid mineral medium, such as paraffin urea minimal (PUM), paraffin gelatin minimal (PGM), gelatin minimal (GM), and GM agar (GMA) slants containing only simple sources of C and N were used, just like the human and the armadillo isolates of these organisms reported earlier . Morphologically, metabolically and enzymologically, these were closely related to the previous ones and were also chemoautotrophic in nature . Serologically there appears to be a heterogenicity in these isolates, i.e., some of them showing higher affinity to nocardioforms while others showing significant binding to several mycobacteria . Normal (uninfected) mouse foot-pad harvests were not found to harbour such organisms.

Indian Pediatr, 1990 Apr, 27(4), 327 - 32
A feasible strategy for a health care package; Ghosh S; PIP: The National Health Policy adopted in India in 1982 proposed primary health care involving the community and relying on maternal-child health (MCH) services . The Bhore Committee stressed similar priorities in 1946, but implementation was deficient . In 1988 there were 100.000 sub-centers (SC), 15.000 primary health centers (PHC), and 800 community health centers (CHC) . The 7th 5-Year Plan envisioned a village health guide (VHG) and trained worker for 1000 people . A SC was to serve 5000 people with 2 health workers, and a PHC to care for 30.000 people . The budget increased for these efforts and for the universal immunization program (UIP) . Antinatal care coverage for women was 60-75%, while tetanus toxoid coverage was complete (100%) . Still only a low percentage of women utilized antenatal care: 1.4-32.8% . Nonutilization was due to traditional beliefs, emphasis of tubectomy and immunization targets, and low rate of registration by PHCs of pregnant women . Immunization coverage exceeded 40% in some states, thus it could be optimal for the administration of 2 doses of tetanus toxoid, for antenatal examination (to identify high-risk mothers, anemia, and toxemia), and for giving iron and folic acid tablets (60 mg elemental iron and 500 mg folic acid per tablet to reduce the 60-70% rate of anemia) . A national nutrition study showed that hemoglobin below 8 g/dl and maternal weight below 40 kg at 29 weeks of pregnancy was strongly associated with low birth weight . Trained traditional birth attendants (TBAs) could identify causes for hospital referral . At 4-6 months of the baby's life semisolids enter the diet, and family planning advice can be dispensed . Agenda include during UIP contact at 6-8 weeks: diphtheria (DPT), oral poliovirus vaccine (OPV), and tuberculosis (bacillus Calmette-Guerin or BCG) shots; health assessment; and teaching of breast feeding to primipara . One vitamin A dose at 1 year and another at 1 and 1/2 year of life during DPT and OPV booster shots can be administered . Flash cards and illustrations facilitate communication for health workers . Hand washing after cleaning the baby should be stressed . Additional inputs along with UIP can lead to improved maternal and child care .

Antibiot Khimioter, 1990 Apr, 35(4), 14 - 6
{Effect of the producer type on the nature of antibiotic fermentation}; Rudakova AV et al.; Fermentation processes in production of bacitracin, a polypeptide antibiotic by Bacillus licheniformis, and oleandomycin, a macrolide antibiotic by Streptomyces antibioticus, were studied comparatively . It was shown that the antibiotic-producing actinomycete was characterized by a prolonged phase of growth retardation . The highest efficiency of the control actions was observed at the beginning of the fermentation . They were aimed at intensifying the substrate usage during the growth phase and activation of cell metabolism . Controlled cultivation of the Bacillus representative was based on its capacity of achieving the maximum growth rate possible under the certain conditions . Therefore, an increase in the quantity of the synthesized antibiotic was due, under such conditions, to inhibition of the culture growth by various means including lower mass exchange intensity.

Proc Natl Acad Sci U S A, 1990 Apr, 87(8), 2960 - 4
Use of a tuberculin purified protein derivative--Asn-Ala-Asn-Pro conjugate in bacillus Calmette-Guérin primed mice overcomes H-2 restriction of the antibody response and avoids the need for adjuvants; Lussow AR et al.; Because of its immunodominancy, and because it is conserved in different geographical isolates of Plasmodium falciparum, the repetitive sequence of the circumsporozoite protein, (Asn-Ala-Asn-Pro)n {(NANP)n}, has been envisaged for the development of an anti-falciparum malaria subunit vaccine . However, the murine immune response to (NANP)n peptides, either carrier-free or coupled to carrier proteins, was shown to be inducible only by using strong (e.g., Freund's) adjuvants . Furthermore, response to the carrier-free peptide, administered in adjuvant, is genetically restricted to I-Ab mice . In the present paper, we report that high titers of antibodies against the NANP repetitive epitope were obtained in responder C57BL/6 (H-2b) mice when they were primed with live BCG (bacillus Calmette-Guerin Mycobacterium tuberculosis var . bovis) and immunized once with the synthetic peptide (NANP)40 coupled to tuberculin purified protein derivative (PPD) without the use of any adjuvant . This approach also led to the production of high titers of anti-NANP antibodies in ASW (H-2s), B10.RIII (H-2r), BALB/c (H-2d), C3H/He (H-2k), and DBA/1 (H-2q) nonresponder mice after two injections of the conjugate . In both cases, BCG priming was obligatory for the induction of antibodies reacting with the synthetic peptide . The levels of anti-NANP antibodies in nonresponder BALB/c mice were demonstrated to be comparable to the levels induced after PPD-(NANP)40 immunization in Freund's complete or incomplete adjuvant . The antibodies induced were also capable of recognizing P . falciparum sporozoites in immunofluorescence assays and, furthermore, these antibodies inhibited the penetration of live sporozoites into human hepatocytes in vitro . This system functioned independently of the subjects' resistance or susceptibility to BCG infection . Given the widespread natural exposure to mycobacterial antigens and the extensive use of BCG and PPD in the human population, this approach might be envisaged for vaccination with malaria peptides.

J Bacteriol, 1990 Apr, 172(4), 2160 - 7
Synthesis and secretion of a Bacillus circulans WL-12 1,3-1,4-beta-D-glucanase in Escherichia coli; Bueno A et al.; The synthesis and secretion of a 1,3-1,4-beta-D-glucanase were studied in different strains of Escherichia coli transformed with plasmids carrying the Bacillus circulans WL-12 1,3-1,4-beta-D-glucanase structural gene . This gene (named BGC) is contained within a 1.9-kilobase BamHI-HindIII fragment and directs the synthesis in E . coli of an enzyme that specifically degrades lichenan . Only one active form of the enzyme was found when the gene was expressed in different E . coli strains . The electrophoretic pattern of this protein showed a molecular weight that was approximately the same as that of the mature beta-glucanase secreted from B . circulans WL-12, suggesting that the processing of this protein may be similar in both species . As deduced from maxicell experiments, the Bacillus parental promoter directs the synthesis in E . coli . Pulse-chase experiments showed that the protein may be cotranslationally processed.

J Clin Oncol, 1990 Apr, 8(4), 608 - 14
Adjuvant bacillus Calmette-Guérin therapy in non-Hodgkin's malignant lymphomas: long-term results of a randomized trial in a single institution; Ravaud A et al.; Between 1973 and 1977, 48 patients less than 65 years old with non-Hodgkin's malignant lymphoma (NHML) of poor prognosis (+/- high grade malignancy, +/- clinical stages III or IV, +/- first or repeated relapse) were included in a prospective clinical trial . After complete remission (CR), obtained with chemotherapy and radiotherapy, patients were randomized to receive bacillus Calmette-Guerin (BCG) or no further therapy . BCG was administered in weekly scarifications up to 3 years . Forty-three patients are assessable . Twenty-four patients have relapsed: nine out of 21 in the BCG group, and 15 out of 22 in the control group . There is a significant difference in favor of the BCG group in disease-free survival (P = .03) . Twenty-one patients have died, 18 from NHML: seven in the BCG group, and 11 in the control group . There is a significant difference in favor of the BCG group for overall survival at 10 years (P = .05) . A multivariate analysis points out BCG as a significant prognostic factor . Adjuvant BCG may improve particularly disease-free survival and overall survival for patients with clinical stages I and II or intermediate- and high-grade malignancy . These results suggest that in patients less than 65 years old with NHML of poor prognosis, BCG may significantly increase disease-free survival and overall survival.

Immunol Cell Biol, 1990 Apr, 68 ( Pt 2), 87 - 93
Antibody against Bacillus thuringiensis phosphatidylinositol-phospholipase C: some examples of its potential uses; Theveniau MA et al.; We have purified the phosphatidylinositol-phospholipase C enzyme from the bacterium Bacillus thuringiensis . This enzyme is able to release in soluble form molecules which are anchored to membranes via a glycan-phosphatidylinositol group . It exhibits a molecular weight of 33-35 kDa . We raised polyclonal antisera against the molecule and used them in immunoblot as well as radioimmunoassays for enzyme detection . This last technique should facilitate monitoring of chromatographic steps during enzyme purification . We coupled antibodies to Sepharose beads in order to remove the enzyme from incubation media . This reagent also proved to be particularly useful in control experiments designed to ascertain that the observed release of molecules is due to the action of the phosphatidylinositol-phospholipase C enzyme and not to spontaneous release or to cleavage by nonspecific hydrolases . A search for cross-reactive molecules in other bacterial strains or mammalian tissues gave negative results . This leads to the conclusion that a great diversity exists between phosphatidylinositol-phospholipases C, even among different bacterial strains.

Appl Environ Microbiol, 1990 Apr, 56(4), 1128 - 34
Expression of Bacillus thuringiensis delta-endotoxin genes during vegetative growth; Mettus AM et al.; Bacillus thuringiensis delta-endotoxin (crystal protein) genes are normally expressed only during sporulation . It is possible to produce crystal protein during vegetative growth by placing B . thuringiensis crystal protein genes downstream of a strong vegetative promoter . By removing a possible transcriptional terminator of the tetracycline resistance gene of pBC16 and inserting a multiple cloning site, delta-endotoxin genes can be cloned downstream from the tetracycline resistance gene promoter . This construct allows for readthrough transcription from the strong vegetative promoter . Crystal protein is then produced during vegetative growth as well as during sporulation in both B . thuringiensis and Bacillus megaterium . This construct also allows for production of delta-endotoxin in B . thuringiensis strains that do not normally produce delta-endotoxin because of a defect in sporulation.

Proc Natl Sci Counc Repub China B, 1990 Apr, 14(2), 69 - 74
The rabbit muscle phosphofructokinase gene: cDNA cloning and sequencing; Kao MC et al.; A partial cDNA for rabbit muscle phosphofructokinase (Rm-PFK) has been cloned and sequenced . The nucleotide sequence of the cDNA agrees with the previously determined Rm-PFK genomic sequence . In addition, the amino acid sequence deduced from the cDNA is nearly identical to the Rm-PFK sequence previously determined by peptide analysis . A significant degree of homology exists when the amino acid sequence of Rm-PFK is compared with the sequences of Bacillus stearothermophilus PFK or Escherichia coli PFK-1 . The cloning and sequencing of the PFK cDNA fragment represents significant progress toward the long term goal of using site-specific mutagenesis to investigate the structure-function relationships in this allosteric enzyme.

Rev Ig Med Muncii Med Soc Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Pneumoftiziol, 1990 Apr-Jun, 39(2), 157 - 72
{Pleuropulmonary decortication in the treatment of chronic pleural sacs}; Finichiu DG et al.; The paper reports on the indications and results obtained, in the study of 224 autonomous or associated pleural pulmonary decortications . The decortication is aimed at removing the chronic pleural sack and the possible parenchymatous lesions and at the recovery of the maximum functional pulmonary parenchyma . For the recently chronic cases, the authors suggest the early surgical indication; the pleural pulmonary decortication is better achieved technically and gives better lesional and functional results if it is made earlier; the optimum operatory moment of the decortication superimposes on the moment of chronicization of the pleural sack (2-6 months) . The decortications for chronic pleural tuberculous sacks were followed by good results in 85.72% of cases, and those for nontuberculous affections in 95.23% . The mortality for the whole sample was of 2.23% . Nowadays, when empyemas generated by old methods of medico-surgical collapse and the chronic bacillary pleurisies are more and more seldom, the pulmonary decortication seems to improve its lesional results, and mainly to give better functional results.

Rev Ig Med Muncii Med Soc Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Pneumoftiziol, 1990 Apr-Jun, 39(2), 127 - 33
{The risk of tuberculous infection on the territory of Iaşi County in a 5-year period}; Savin L et al.; The authors study the infection risk (IR) in the Iasi County, between 1983 and 1987, at the age of 6-7 years old, in relation to the evolution of tuberculosis incidence in children and to the bacillary incidence and prevalence . TR was maintained, in the period of study, at about 0.90%, thus corresponding to the bacillary incidence on the territory, on average of 39% inhabitants . Calculation of the contagiousness parameter of the contaminating sources showed that a source is infectious for about 13 children at 100,000 inhabitants . The findings above plead for the necessity of improving the county fighting program against tuberculosis.

Appl Environ Microbiol, 1990 Apr, 56(4), 963 - 70
L-lysine production at 50 degrees C by mutants of a newly isolated and characterized methylotrophic Bacillus sp; Schendel FJ et al.; The amino acid L-lysine was produced from homoserine auxotrophic and S-(2-aminoethyl)-L-cysteine-resistant mutants of a newly isolated gram-positive methylotrophic bacterium, capable of growth on methanol at 60 degrees C . The temperature optimum for growth was between 50 and 53 degrees C . These aerobic, gram-positive, endospore-forming, rod-shaped bacteria required biotin and vitamin B12 for growth . Extracts of the bacteria grown on methanol lacked hydroxypyruvate reductase and contained hexulose 6-phosphate synthase activity . Therefore, these bacteria were considered to be type I methylotrophic bacteria of the genus Bacillus . Fed-batch fermentations resulted in cell densities of 50 g of cell dry weight per liter . Biomass yields on carbon, nitrogen, phosphate, and sulfate were determined . Generation of homoserine auxotrophic and amino acid analog-resistant mutants resulted in L-lysine concentrations of nearly 20 g/liter in fed-batch fermentations.

Rev Med Chir Soc Med Nat Iasi, 1990 Apr-Jun, 94(2), 329 - 36
{The present-day aspects of urogenital tuberculosis . Comments on 113 cases}; Filimon C et al.; One hundred and thirteen new cases of urogenital tuberculosis, admitted and treated in the interval 1980-1986 (an average number of 16 cases yearly) are reported . The analysis of these patients includes age, sex, bacillary history, symptoms, clinical forms, diagnostic methods, therapeutical management and late results . Seven cases are detailed . It is concluded that the treatment with the actual tuberculostatic drugs is efficient provided the urogenital tuberculosis is early detected . In case this disease is late detected the treatment is associated, drug and surgical.

Agric Biol Chem, 1990 Apr, 54(4), 999 - 1005
Purification and some properties of dipeptidyl carboxypeptidase from Bacillus pumilus; Nagamori Y et al.; An intracellular protease from a bacterium, Bacillus pumilus HL721, was purified about 5000-fold by chromatography with a Q-Sepharose Fast Flow column, TSK-gel HA-1000 glass column, and TSK-gel G3000SWXL column using Bz-Gly-Ala-Pro as a substrate . The enzyme was the most active at pH around 7.5 and stable from 4.5 and 8.0 . The enzyme activity was inhibited by Cu2+, EDTA, N-ethylmaleimide, o-phenanthroline, and p-chloromercuribenzoic acid . The molecular weight of the enzyme was 155,000 by gel filtration . The enzyme removed dipeptide from the carboxyl end of some peptides used as substrates . From these results the enzyme seems to be a dipeptidyl carboxypeptidase.

Enzyme Microb Technol, 1990 Apr, 12(4), 242 - 54
Bioconversions in aqueous two-phase systems; Andersson E et al.; Bioconversions involving enzymes and/or microbial cells in aqueous two-phase systems are reviewed . The partitioning of biocatalysts, substrates, and products is discussed in relation to their size . The efficiency of retaining biocatalysts in aqueous two-phase systems is summarized in relation to other methods of recirculating . The influence of phase components on the activity and the stability of enzymatic biocatalysts is exemplified with penicillin acylase and the cellulolytic enzyme system, and the effect of phase components on biocatalytic living cells is exemplified with the production of alpha-amylase with Bacillus sp . Process design costs in bioconversions in aqueous two-phase systems are briefly summarized.

Biochim Biophys Acta, 1990 Mar 29, 1038(1), 29 - 38
Electron transferase activity of diaphorase (NADH: acceptor oxidoreductase) from Bacillus stearothermophilus; Matsue T et al.; Electrochemical kinetic measurements were carried out for electron-transfer between NADH and the oxidized forms of mediators (ferrocenylmethanol (FMA), ferrocenyl-1-ethanol (FEA), N,N,N',N'-tetramethylphenylenediamine (TMPD), Co(Phen)2+(3) and Fe(CN)4-(6)) catalyzed by diaphorase (NADH: acceptor oxidoreductase, EC 1.6.99.-) purified from Bacillus stearothermophilus . Cyclic voltammograms for the mediators with excess NADH in the presence of diaphorase gave steady-state currents . The quantitative analysis of the dependence of the current on the mediator concentration yielded a Michaelis constant (Km) and molecular activity (ko), which are difficult to determine by the conventional spectrophotometric method . Small Km and large ko values were observed for the oxidized forms of FMA, FEA and TMPD compared to those for Co(Phen)3+(3) and Fe(CN)3-(6) . It is suggested that the reaction pocket of the present diaphorase is hydrophobic . The present electrochemical procedure for the determination of the kinetic parameters is applicable widely to similar enzyme reactions.

Carbohydr Res, 1990 Mar 25, 197, 227 - 35
Purification and properties of an oligo-1,6-D-glucosidase from an alkalophilic Bacillus species; Yamamoto M et al.; An isomaltose-hydrolyzing alpha-D-glucosidase from the alkalophilic Bacillus designated strain F5 was purified to an electrophoretically homogeneous state . The molecular weight of the purified glucosidase was 60,000 by SDS-poly(acrylamide) gel electrophoresis, and 63,000 by Sephacryl S-200 gel-filtration chromatography . The enzyme was most active for isomaltose at pH 6.0-6.5 and 45 degrees, and stable up to 50 degrees at pH 7.0 and in the range of pH 6.0-9.0 at 50 degrees by 10-min incubation . The apparent Vmax and Km values for isomaltose were 34.5 mumol.min-1.mg-1 of protein, and 3.33 mM . Panose and isomaltotriose are the best substrates for this enzyme . The restricted substrate specificity indicated the assignment of the enzyme to be an oligo-1,6-glucosidase (dextrin 6-alpha-glucanohydrolase; EC 3.2.1.10), but it was suggested that it could be a new type of oligo-1,6-glucosidase on the basis of its action on a series of (1----4)-alpha-malto-oligosaccharides.

Lancet, 1990 Mar 24, 335(8691), 707 - 10
Immunisation practice in developed countries; Hinman AR et al.; PIP: Immunization practice in 32 countries in Europe, North America, Japan, and Australia is reviewed . in most countries, immunization practices are set by the federal government which sometimes works with the private sector . Almost all countries routinely immunize against diphtheria, tetanus, whooping cough, polio, and measles . About half try to prevent rubella, several try to prevent mumps, usually in combination with measles and rubella (MMR) . More than half use bacillus Calmette-Guerin (BGG) vaccine to prevent tuberculosis, and a few give Hemophilus Influenza type B polysaccharide . Poliomyelitis vaccine comes in 2 forms: 1) oral live attenuated (OPV) or injectable inactivated (IPV) . OPV is more used, but there is a new "enhanced potency IPV." All countries except Japan give DPT in 3 doses during the 1st year of life . OPV is usually given at the same time that DPT is . Measles vaccine or MMR is usually given between 12 and 18 months of age . Primary vaccine failure occurs in 2-5% of people who get measles vaccine, but this may be enough to "sustain transmission." In most countries, the government provides for immunizing children . An exception in the US . In the UK, low coverage has taken place because of concern for adverse reactions (whooping cough) or lack of appreciation of the disease's impact (measles) . Coverage against both measles and pertussis has improved in the UK lately . In each developed country, vaccines have had "spectacular" effects . However, there are too many contraindications and there is "undue fear of adverse events." Also, there are surveillance deficiencies, a lack of coordination, and countries vary in their commitment to "reduction/elimination targets." Varicella vaccine, respiratory syncytial virus vaccine, and rotavirus vaccine are being considered for universal use . Attempts are being made to improve the safety of some vaccine .

Am J Med, 1990 Mar 23, 88(3C), 34S - 37S; discussion 38S-42S
Gram-negative bacillary pneumonia in the nosocomial setting . Role of aztreonam therapy; Cook JL; Gram-negative bacterial pneumonia is the leading cause of fatal nosocomial infection in this country . Predisposing factors include altered upper respiratory tract flora and altered barriers that normally protect the sterile lower respiratory tract from invasion by pharyngeal bacteria . Aztreonam, which is highly active against most gram-negative pathogens and which does not cause nephrotoxicity, has been evaluated in the treatment of nosocomial pneumonia . In vitro and pharmacokinetic data on aztreonam indicate that this agent provides an alternative agent for use when resistance to cephalosporin and aminoglycoside antibiotics has developed . Data further suggest that aztreonam may interact synergistically with aminoglycosides against gram-negative pathogens . Clinical study supports the usefulness of aztreonam against gram-negative nosocomial pneumonia . Since aztreonam is inactive against gram-positive and anaerobic bacteria, it must be used in combination with other antibiotics when these pathogens are suspected.

Biochemistry, 1990 Mar 20, 29(11), 2747 - 57
Phospholipids chiral at phosphorus . Stereochemical mechanism of reactions catalyzed by phosphatidylinositide-specific phospholipase C from Bacillus cereus and guinea pig uterus; Lin GL et al.; (Rp)- and (Sp)-1,2-dipalmitoyl-sn-glycero-3-thiophosphoinositol (DPPsI) were synthesized as a mixture and their configurations assigned on the basis of the stereospecific hydrolysis catalyzed by phospholipase A2 (PLA2) from bee venom . PLA2 is known to be stereospecific to the Rp isomer of 1,2-dipalmitoyl-sn-glycero-3-thiophosphocholine (DPPsC) and 1,2-dipalmitoyl-sn-glycero-3-thiophosphoethanolamine (DPPsE) . Since the configurations of (Rp)- and (Sp)-DPPsI correspond to those of (Sp)- and (Rp)-DPPsC, respectively, due to a change in priority, the isomer specifically hydrolyzed by PLA2 was assigned to (Sp)-DPPsI . The DPPsI analogues were then used to probe the mechanism and to elucidate the steric course of the reaction catalyzed by phosphatidylinositide-specific phospholipase C (PI-PLC) from Bacillus cereus and for both isozyme I and isozyme II of PI-PLC from guinea pig uterus . It was found that the Rp isomer of DPPsI is the preferred substrate for all three PI-PLCs . Thus PI-PLC shows the same stereospecificity as phosphatidylcholine-specific PLC (PC-PLC), which prefers the Sp isomer of DPPsC . The ratio of the two products inositol 1,2-cyclic phosphorothioate (cIPs) and inositol phosphorothioate (IPs) was not significantly perturbed by the use of phosphorothioate analogue for all three PI-PLCs, which implies that IPs is not produced by enzyme-mediated ring opening of cIPs and supports a parallel pathway for the formation of both products . In order to elucidate the steric course of the cyclization reaction, exo and endo isomers of cIPs were synthesized and their absolute configurations at phosphorus were determined by nuclear magnetic resonance and other techniques . It was found that exo-cIPs is the product produced by all three PI-PLCs . Thus the steric course of the conversion DPPsI to cIPs catalyzed by all three PI-PLCs was inversion of configuration at phosphorus . These results taken together suggest that the reaction catalyzed by PI-PLC most likely proceeds via direct attack by the 2-OH group to generate the cyclic product, and parallelly by water to generate the noncyclic inositol phosphates, without involving a covalent enzyme-phosphoinositol intermediate.

Biochem Biophys Res Commun, 1990 Mar 16, 167(2), 402 - 6
Identification of germination gene of Bacillus megaterium; Tani K et al.; Glucose, KNO3, proline and leucine initiate the spore germination of B . megaterium ATCC 12872, but not of B . megaterium ATCC 19213 . In order to isolate the gene concerning germination of B . megaterium ATCC 12872, we constructed its gene library in plasmid vector, and introduced into B . megaterium ATCC 19213 . We obtained a transformant whose spores differed from those of the wild type strain with respect to germinability . Spores of this transformant could be germinated by glucose, proline or leucine . The recombinant plasmid prepared from this transformant was found to carry 2 kilobase pairs fragment of B . megaterium ATCC 12872 DNA . This fragment may contain the gene encoding the protein which plays an important role in germination.

Biochem J, 1990 Mar 15, 266(3), 933 - 6
Loss of alkalophily in cell-wall-component-defective mutants derived from alkalophilic Bacillus C-125 . Isolation and partial characterization of the mutants; Aono R et al.; The cells of alkalophilic Bacillus sp . C-125 are shaped by peptidoglycan and enclosed by two acidic polymers (teichuronic acid and teichuronopeptide), which bind to the peptidoglycan . Three kinds of mutant strains defective in these acidic polymers were isolated from the strain C-125 . These mutants grow poorly at alkaline pH to extents related to the degree of defect in the polymers, suggesting that these acidic polymers are essential for growth in an alkaline environment . These polymers may diminish penetration of hydroxide ions.

Eur J Biochem, 1990 Mar 10, 188(2), 355 - 9
Stereochemistry and lifetime of the GTP hydrolysis intermediate at the active site of elongation factor Tu from Bacillus stearothermophilus as inferred from the 17O-55Mn superhyperfine interaction; Kalbitzer HR et al.; Electron paramagnetic resonance spectroscopy has been used to obtain information on the structure and stability of the products of GTP cleavage at the active site of elongation factor Tu (EF-Tu) from Bacillus stearothermophilus . Using stereospecifically labelled (Sp)-(Rp)-{beta-17O}GTP (prepared by modification of a previously published procedure which is now also suitable for guanine nucleotides), it was found that only one of the two possible diastereomers (Sp) led to detectable line-broadening of the EPR spectrum of Mn2+ at the active site of EF-Tu (linewidth 1.5 mT), whereas the Rp isomer caused the same linewidth as unlabelled nucleotide (1.3 mT) . From our earlier work and from a demonstration that the lifetime of the state giving the broadened spectrum is too long to be assigned to the EF-Tu.GDP.Mn complex {the rate constant for decay as measured by displacement of GDP by the fluorescent 2'(3')-O-(N-methylanthraniloyl)-GDP is 6.2 x 10(-3) s-1 at 25 degrees C and pH 6.8}, we conclude that the broadened signal arises from the EF-Tu.Mn.GDP.Pi complex, the predominant steady-state species . During the hydrolysis of GTP the Mn2+ remains bound to the beta-phosphate oxygen of GDP which arises from the beta pro-S oxygen of GTP, possibly until GDP dissociates and certainly until Pi dissociates . Addition of elongation factor Ts (EF-Ts) to this intermediate leads to rapid reduction of the linewidth to that expected for random distribution of interactions of one 17O and two 16O atoms of GDP with Mn2+, and is not distinguishable from that exhibited by (Rp)-{beta-17O}GTP in the corresponding complex in the presence of EF-Ts.

Eur J Biochem, 1990 Mar 10, 188(2), 213 - 8
Asymmetric distribution of the phosphatidylinositol-linked phospho-oligosaccharide that mimics insulin action in the plasma membrane; Varela I et al.; We have investigated the topography of a glycosyl-phosphatidylinositol implicated in insulin action by a combination of two complementary methods: (a) chemical labelling with a non-permeable (isethionyl acetimidate) and a permeable (ethyl acetimidate) probe; and (b) enzymatic modifications with beta-galactosidase (EC 3.2.1.23) or phosphatidylinositol-specific phospholipase C (EC 3.1.4.3) . Using the first approach the majority of the glycosyl-phosphatidylinositol is found in the outer surface of intact hepatocytes, adipocytes, fibroblasts and lymphocytes, but not in erythrocytes which presented only a 20% of the total labelled glycosyl-phosphatidylinositol to the exterior . Upon insulin addition (10 nM), about 60% of the total glycosyl-phosphatidylinositol was hydrolysed in both hepatocytes and adipocytes but not in erythrocytes . In agreement with the extracellular localization in hepatocytes and with the proposed role of this glycolipid in insulin action, treatment of rat hepatocytes with beta-galactosidase from Escherichia coli, an enzyme that hydrolyses the oligosaccharide moiety of the glycosyl-phosphatidylinositol, cleaved 65% of the total glycophospholipid and blocked the effect of insulin (but not of glucagon) on pyruvate kinase (EC 2.7.1.40) . Similar treatment with phosphatidylinositol-specific phospholipase C from Bacillus cereus hydrolysed 62% of the total glycosyl-phosphatidylinositol . From the various approaches used it is concluded that the majority of this glycophospholipid is at the outer surface in a variety of insulin-sensitive cells.

Eur J Biochem, 1990 Mar 10, 188(2), 275 - 81
Conformational and thermodynamic effects of naturally occurring base methylations in a ribosomal RNA hairpin of Bacillus stearothermophilus; Heus HA et al.; The 3'-terminal colicin fragments of 16S ribosomal RNA were isolated from Bacillus stearothermophilus and from its kasugamycin-resistant (ksgA) derivative lacking N6-dimethylation of the two adjacent adenosines in a hairpin loop . The fragment from the ksgA strain still contains a naturally occurring N2-methylguanosine in the loop . An RNA molecule resembling the B . stearothermophilus colicin fragment but without modified nucleosides was synthesized in vitro using a DNA template and bacteriophage T7 RNA polymerase . Proton-NMR spectra of the RNAs were recorded at 500 MHz . The imino-proton resonances of base-paired G and U residues could be assigned on the basis of previous NMR studies of the colicin fragment of Escherichia coli and by a combination of methylation-induced shifts and thermal melting of base pairs . The assignments were partly confirmed by NOE measurements . Adenosine dimethylation in the loop has a distinct conformational effect on the base pairs adjoining the loop . The thermal denaturation melting curve of the enzymatically synthesized RNA fragment was also determined and the transition midpoint (tm) was found to be 73 degrees C at 15 mM Na+ . A comparison with previously determined thermodynamic parameters for various colicin fragments demonstrates that base methylations in the loop lead to a relatively strong destabilization of the hairpin helix . In terms of free energy the positive contribution of the methylations are in the order of the deletion of one base pair from the stem . Other data show that recently published free-energy parameters do not apply for certain RNA hairpins.

J Bacteriol, 1990 Mar, 172(3), 1539 - 46
Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae; Bowler C et al.; Recombinant clones containing the manganese superoxide dismutase (MnSOD) gene of Bacillus stearothermophilus were isolated with an oligonucleotide probe designed to match a part of the previously determined amino acid sequence . Complementation analyses, performed by introducing each plasmid into a superoxide dismutase-deficient mutant of Escherichia coli, allowed us to define the region of DNA which encodes the MnSOD structural gene and to identify a promoter region immediately upstream from the gene . These data were subsequently confirmed by DNA sequencing . Since MnSOD is normally restricted to the mitochondria in eucaryotes, we were interested (i) in determining whether B . stearothermophilus MnSOD could function in eucaryotic cytosol and (ii) in determining whether MnSOD could replace the structurally unrelated copper/zinc superoxide dismutase (Cu/ZnSOD) which is normally found there . To test this, the sequence encoding bacterial MnSOD was cloned into a yeast expression vector and subsequently introduced into a Cu/ZnSOD-deficient mutant of the yeast Saccharomyces cerevisiae . Functional expression of the protein was demonstrated, and complementation tests revealed that the protein was able to provide tolerance at wild-type levels to conditions which are normally restrictive for this mutant . Thus, in spite of the evolutionary unrelatedness of these two enzymes, Cu/ZnSOD can be functionally replaced by MnSOD in yeast cytosol.

Zhonghua Yu Fang Yi Xue Za Zhi, 1990 Mar, 24(2), 93 - 5
{The effects of six kinds of insecticides on gamasid mites}; Zhou HF; The present paper reports on the results of six kinds including 14 chemical or biological insecticides on the gamasid mites . (1) Using organophosphorus compounds such as quinothion, supracide, bromochlophus, sumithion at a dose of 0.2g per m2, at a dose of 1 g per m2 abate, and malathion at a dose of 49 per m2, they were all sufficient to produce 100% mortality on the mites Eulaelaps stabularis; trichlorphon at a dose of 4g per m2 produced 96.4% kill of the mites . (2) Organochloride insecticide BHC containing 93% gamma HCH was not effective on the same mites . (3) at a dose of 0.1 g per m2 of pyrethroid; decis, all the Eulaelaps stabularis were knocked down immediately, but 87% of the mites recovered in 24 hours . (4) The acaricides and cycloprate, the molt-inhibiting hormone Su-urea No . 1 and No . 2, and Bacillus thuringiensis var, Isralensis had not effect on the mites Eulaelaps stabularis and/or Haemolae laps glasgowi, It is suggested that organophosphorus compounds should the insecticide of choice against the gamasid mites.

Nippon Hinyokika Gakkai Zasshi, 1990 Mar, 81(3), 425 - 32
{Intravesical bacillus Calmette-Guerin instillation therapy in superficial bladder cancers--clinical study on prognostic factors}; Shinka T et al.; Intravesical instillation of Tokyo 172 strain bacillus Calmette-Guerin (BCG) was performed on 137 patients with superficial bladder cancer (Ta and T1) after transurethral tumor resection as a prophylaxis against tumor recurrence . The recurrence rate of tumors was compared with that of historical controls and was estimated by the person-years method . There were statistically significant decreases in recurrent tumors following BCG therapy . To clarify the efficacy of intravesical BCG therapy, prognostic significance of several factors were evaluated in 90 patients with initial bladder cancer treated with TUR and BCG instillation, and compared to those of controls . Prophylactic effects were statistically better for those with multiple tumors, grade 3 lesions or Ta lesions than for control patients . No correlation between purified protein derivative responsiveness and favorable results could detected . There were no marked side effects or fatal complications of BCG therapy during the observation periods . Our results suggest that BCG is able to change the biological behavior of superficial bladder cancers with multiple, high grade or low stage lesions . The intravesical BCG instillation seems to be effective and safe as a prophylaxis against the recurrence of superficial bladder tumors.

FEMS Microbiol Lett, 1990 Mar 1, 56(1-2), 97 - 104
Directed mutagenesis of selected regions of a Bacillus thuringiensis entomocidal protein; Ahmad W et al.; Comparison of the sequences of Bacillus thuringiensis entomocidal toxins of widely differing specificity reveals six conserved domains . The role of one of the most highly conserved domains (D1) located near the N-terminus has been investigated by site directed mutagenesis at two positions . Although preliminary results indicate that the capacity of the mutants to bind to putative receptors on the plasma membrane of susceptible cells was unaffected, toxicity in vivo was reduced by 70-80% . The role of the highly hydrophobic segment exposed at the N-terminus of the toxin after proteolytic activation was investigated by substituting two aspartate residues for phenylalanine and valine located adjacent to each other in the centre of this segment . The toxicity of the resulting mutant protein was only 40% of the unmutated toxin but again preliminary results suggest that binding to putative receptors was unaffected . These results suggest that regions close to the N-terminus of this and similar toxins may play an important role in the membrane insertion event which is believed to follow receptor recognition.

J Am Mosq Control Assoc, 1990 Mar, 6(1), 43 - 6
Field efficacy of Vectobac-12AS and Vectobac-24AS against black fly larvae in New Brunswick streams (Diptera: Simuliidae); Riley CM et al.; The control of black fly larvae using 2 aqueous suspension formulations of Bacillus thuringiensis, serotype H-14 was evaluated in 6 trials on small and large sized streams in New Brunswick . Applications equivalent to 25 ppm (Vectobac-12AS) or 12.5 ppm (Vectobac-24AS) over a 1-min period resulted in 100% control of black fly larvae up to 3 km downstream of the application point . Over the 6 trials, effective carry was seen to increase with increased stream discharge which varied from as low as 0.8 m3/min in the small stream to 108.6 m3/min in the large stream.

J Am Mosq Control Assoc, 1990 Mar, 6(1), 144 - 7
Salt marsh mosquito control in Portsmouth, Rhode Island; Christie GD; The Portsmouth, Rhode Island Mosquito Control Program used granular Bacillus thuringiensis var . israelensis to successfully control salt marsh mosquitoes in a 9 acre salt marsh while monitoring for, planning and implementing small scale open marsh water management in the marsh . Single season larviciding costs were roughly 4% that of contracted open marsh water management in this marsh.

Am J Trop Med Hyg, 1990 Mar, 42(3), 215 - 21
An epidemic of Oroya fever in the Peruvian Andes; Gray GC et al.; Between February and October 1987, a febrile illness killed 14 persons and seriously affected at least 14 others in Shumpillan, a remote Peruvian mountain village of 353 people . The illness was characterized by fever, headache, chills, and pallor . The fatality rate of untreated cases was 88% . The patients, 71% of whom were male, were 1-75 years of age . Fatal illnesses progressed from lethargy to coma to death in 3-60 days . Patients treated empirically with chloramphenicol survived . Bartonella bacilliformis was isolated from the whole blood of 3 patients . A serologic study revealed a high prevalence of antibodies to B . bacilliformis in the villagers . It is concluded that the villagers suffered from an epidemic of Oroya fever.

J Bacteriol, 1990 Mar, 172(3), 1485 - 90
Chloride transport pathways and their bioenergetic implications in the obligate acidophile Bacillus coagulans; McLaggan D et al.; The protonophore-mediated collapse of the large delta pH that acidophiles maintain across their cytoplasmic membranes was augmented by the presence of Cl-, and Cl- influx into the cells occurred evidently in response to the protonophore-induced increase in the inside-positive membrane potential (+ delta psi) . In respiring cells, the addition of Cl- but not SO4(2-) salts caused a rapid and precipitous decrease in the + delta psi . A Nernstian relationship between the imposed transmembrane K+ gradient and the valinomycin-induced K+ diffusion potentials was observed when everted membrane vesicles were loaded with K2SO4 or KH2PO4 but not when loaded with KCl or KNO3 . Thus, electrogenic Cl- transport occurred in Bacillus coagulans . In addition, a nonelectrogenic temperature-sensitive Cl- transport mechanism, with the net Cl- efflux coefficient (PCl-) ranging from 1.5 x 10(-4) to 6.1 x 10(-6) cm/s, accounted for the massive Cl- efflux from Cl(-)-loaded cells . Thus, B . coagulans, despite its dependence on the + delta psi and therefore the need to exclude anions, apparently possesses specific mechanisms for Cl- permeation . Active cells of B . coagulans prevented Cl- accumulation from attaining an electrochemical equilibrium, maintaining a delta micro Cl- of ca . -63 mV . B . coagulans therefore also possesses an energy-dependent mechanism for Cl- exclusion from the cells.

J Bacteriol, 1990 Mar, 172(3), 1312 - 20
Conserved structures of cell wall protein genes among protein-producing Bacillus brevis strains; Ebisu S et al.; Bacillus brevis HPD31 contains a surface (S)-layer protein, termed the HWP, which forms a hexagonal array in the cell wall . The 5' region of the HWP gene was isolated from a DNA library constructed in bacteriophage vector EMBL3 from a partial BamHI digest of the chromosomal DNA . The 3' region contained in a 2.7-kilobase BglII fragment of the DNA was cloned into Escherichia coli, using pUC118 as a vector . On the basis of the chemically determined N-terminal amino acid sequence, the HWP gene was found to encode a polypeptide consisting of 1,087 amino acid residues with a signal peptide of 53 or 23 amino acid residues . The deduced amino acid composition was similar to the chemical amino acid compositions of other S-layer proteins in the predominance of acidic relative to basic amino acids and in the very low content of sulfur-containing amino acids . The deduced amino acid sequence showed high homology (78%) with that of the middle wall protein of B . brevis 47 . Furthermore, the multiple 5' ends of the HWP gene transcripts detected on S1 nuclease analysis closely resembled those of the middle wall protein gene transcripts . This complex structure was also conserved (greater than 85%) in the regulatory regions of two other cell wall protein genes isolated from B . brevis HPD52 and HP033, suggesting that the synthesis of the cell wall proteins is intricately regulated through a similar mechanism in protein-producing B . brevis.

J Am Mosq Control Assoc, 1990 Mar, 6(1), 139 - 40
The effect of Bacillus sphaericus upon the susceptibility of Anopheles quadrimaculatus to Plasmodium berghei; Young MD et al.; Larvae of Anopheles quadrimaculatus were exposed to Bacillus sphaericus . The surviving adults took blood meals on hamsters infected with Plasmodium berghei . Fewer mosquitoes were infected than were the paired controls . The inhibitory action appeared to occur during the early stages of the infection in the mosquito gut.

Microbiol Rev, 1990 Mar, 54(1), 52 - 65
Uncoupler-resistant mutants of bacteria; Krulwich TA et al.; The chemiosmotic model of energy transduction offers a satisfying and widely confirmed understanding of the action of uncouplers on such processes as oxidative phosphorylation; the uncoupler, by facilitating the transmembrane movement of protons or other compensatory ions, reduces the electrochemical proton gradient that is posited as the energy intermediate for many kinds of bioenergetic work . In connection with this formulation, uncoupler-resistant mutants of bacteria that neither exclude nor inactivate these agents represent a bioenergetic puzzle . Uncoupler-resistant mutants of aerobic Bacillus species are, in fact, membrane lipid mutants with bioenergetic properties that are indeed challenging in connection with the chemiosmotic model . By contrast, uncoupler-resistant mutants of Escherichia coli probably exclude uncouplers, sometimes only under rather specific conditions . Related phenomena in eucaryotic and procaryotic systems, as well as various observations on uncouplers, decouplers, and certain other membrane-active agents, are also briefly considered.

J Invertebr Pathol, 1990 Mar, 55(2), 235 - 44
Resistance to Bacillus thuringiensis by the Indian meal moth, Plodia interpunctella: comparison of midgut proteinases from susceptible and resistant larvae; Johnson DE et al.; Midgut homogenates from susceptible and resistant strains of the Indian meal moth, Plodia interpunctella, were compared for their ability to activate the entomocidal parasporal crystal protein from Bacillus thuringiensis . The properties of midgut proteinases from both types of larvae were also examined . Electrophoretic patterns of crystal protein from B . thuringiensis subspecies kurstaki (HD-1) and aizawai (HD-133 and HD-144) were virtually unchanged following digestion by either type of midgut homogenate . Changes in pH (9.5 to 11.5) or midgut homogenate concentration during digestion failed to substantially alter protein electrophoretic patterns of B . thuringiensis HD-1 crystal toxin . In vitro toxicity of crystal protein activated by either type of midgut preparation was equal toward cultured insect cells from either Manduca sexta or Choristoneura fumiferana . Electrophoresis of midgut extracts in polyacrylamide gels containing gelatin as substrate also yielded matching mobility patterns of proteinases from both types of midguts . Quantitation of midgut proteolytic activity using tritiated casein as a substrate revealed variation between midgut preparations, but no statistically significant differences between proteolytic activities from susceptible and resistant Indian meal moth larvae . Inhibition studies indicated that a trypsin-like proteinase with maximal activity at pH 10 is a major constituent of Indian meal moth midguts . The results demonstrated that midguts from susceptible and resistant strains of P . interpunctella are similar both in their ability to activate B . thuringiensis protoxin and in their proteolytic activity.

APMIS, 1990 Mar, 98(3), 244 - 8
Extraction and localization by electron microscopy of an immunosuppressor fraction from Mycobacterium bovis Bacillus Calmette-Guérin (BCG); Hiu IJ; BCG has been used all over the world to immunize against tuberculosis . Nevertheless in certain areas (South India) BCG vaccines failed to show any protective efficacy . Furthermore immunosuppressive cell populations have been reported in experimental mycobacterial infection in mice . The present work reports the localization and isolation of an immunosuppressor fraction from BCG . This lipid fraction called WDB inhibited the skin reactivity of delayed-type hypersensitivity (DTH) to the test antigen CEWA (crystalline egg white albumin) in guinea pigs and depressed the production of immune antibody to SRBC (sheep red blood cells) in mice . WDB is a glycolipid with an approximate mol.wt . of 62,000 . By electron microscopy, WDB was located among the BCG extracellular metabolic products (ECMP) surrounding the BCG cell wall.

J Am Acad Dermatol, 1990 Mar, 22(3), 501 - 12
Bacillary angiomatosis: a newly characterized, pseudoneoplastic, infectious, cutaneous vascular disorder; Cockerell CJ et al.; Bacillary angiomatosis (also called epithelioid angiomatosis) is a newly recognized disease most often characterized by a cutaneous infection with reddish papules of vascular origin . It is caused by a weakly reactive gram-negative bacillus, which can be easily demonstrated in tissue sections with the Warthin-Starry stain . Bacillary angiomatosis usually responds readily to treatment with oral erythromycin, 250 to 500 mg, taken four times a day for 2 weeks to 1 month . Because the skin is the most common site of involvement, it is important that the dermatologist recognize this unusual condition . It is essential that treatment be started as soon as possible because deaths may result from visceral and mucosal involvement . The clinical, histologic, and microbiologic aspects of bacillary angiomatosis are discussed and depicted in detail and speculations regarding the pathogenesis are rendered.

J Antibiot (Tokyo), 1990 Mar, 43(3), 267 - 80
Pumilacidin, a complex of new antiviral antibiotics . Production, isolation, chemical properties, structure and biological activity; Naruse N et al.; New antibiotic pumilacidins A, B, C, D, E, F and G were isolated from the culture broth of a strain of Bacillus pumilus . They are cyclic acylheptapeptide composed of a beta-hydroxy fatty acid, two L-leucine, two D-leucine, L-glutamic acid, L-aspartic acid and L-isoleucine (or L-valine) . Pumilacidin components were inhibitory to herpes simplex virus type 1 and H+, K(+)-ATPase and demonstrated antiulcer activity in rat.

J Invertebr Pathol, 1990 Mar, 55(2), 147 - 51
Influence of media composition on the production of delta-endotoxin by Bacillus thuringiensis var . thuringiensis; Mummigatti SG et al.; Powders of edible leguminous seeds, greengram (Vigna radiata) or soybean (Glycine max), were used as the major protein source with different combinations of soluble starch and/or cane sugar molasses as the major carbohydrate source for the production of delta-endotoxin by Bacillus thuringiensis var . thuringiensis serotype 1 in submerged fermentation . The primary product (lyophilized with 6 g of lactose) yield was 8.7 to 9.1 g/liter from media with dehusked greengram powder and 9.7 to 10.3 g/liter from media with defatted soybean powder in basal medium . The toxicity of primary products was assayed against fifth-instar Bombyx mori larvae by force-feeding . The primary product from the medium containing defatted soybean powder and soluble starch gave a maximum viable spore count of 91.3 x 10(6)/mg, with a corresponding potency of 35,800 IU/mg, whereas the medium containing dehusked greengram powder and cane sugar molasses gave a spore count of 49.5 x 10(6)/mg, with a highest potency of 38,300 IU/mg . Either legume protein in combination with cane sugar molasses yielded primary product 2.1 to 2.4 times more potent than the U.S . standard . The combined carbohydrate source consisting of soluble starch and cane sugar molasses, irrespective of the source of protein in the media, drastically reduced delta-endotoxin production, thereby reducing the potency of the primary products compared to the U.S . standard.

Am J Respir Cell Mol Biol, 1990 Mar, 2(3), 289 - 96
Altered release of eicosanoids by rat alveolar macrophages during granulomatous pulmonary inflammation; Shellito J et al.; Release of arachidonic acid metabolites (eicosanoids) by alveolar macrophages may be important in regulating pulmonary inflammatory reactions . The purpose of this study was to characterize eicosanoids released by rat alveolar macrophages during the evolution of experimentally induced pulmonary inflammation . Immunization with subcutaneous bacillus Calmette-Guerin (BCG) followed 2 wk later by intravenous BCG challenge resulted in mild granulomatous pulmonary inflammation for up to 30 days . At serial intervals, alveolar macrophages were lavaged from the BCG-treated rats as well as from control normal rats . Lavaged macrophages were cultured in vitro, and culture supernatants were assayed by radioimmunoassay for release of prostaglandin E2 (PGE2), Leukotriene B4 (LTB4), and thromboxane B2 (TXB2) . Cells were cultured alone, or with added LPS or calcium ionophore A23187 to stimulate eicosanoid release . During BCG-induced inflammation, spontaneous release of PGE2 and LTB4 was unchanged, while spontaneous release of TXB2 was depressed acutely and then returned to control levels . The capacity of alveolar macrophages to release specific eicosanoids in response to an in vitro stimulus was dramatically altered during the course of BCG-induced inflammation . Stimulated release of PGE2 was transiently increased during acute lung injury, but stimulated release of LTB4 was significantly decreased at all stages of inflammation . Stimulated release of TXB2 was unchanged . These results indicate that during the course of granulomatous pulmonary inflammation there are dynamic changes in the profile of eicosanoids released by alveolar macrophages, both spontaneously and in response to in vitro stimulation . This alteration in the release of eicosanoids by alveolar macrophages may be an important factor in the resolution of pulmonary inflammation.

Bol Oficina Sanit Panam, 1990 Mar, 108(3), 213 - 9
{Control of Simuliidae with an emulsifiable concentrate of Bacillus thuringiensis}; Araujo-Coutinho CJ et al.; The effects of the three flowable concentrate formulations of Bacillus thuringiensis var . israelensis (H-14): Teknar (Zoecon, Sandoz Inc.), Vectobac (Abbott Labs.), and Bactimos (Biochem, Solvay Labs.) against Simulium pertinax were evaluated in 24 streams in the North Littoral Zone of Sao Paulo State, Brazil . There was no significant difference in effective carry (80% or more mortality) among the formulations when applied at the rate of 10 mg/liter for one minute . There was a strong correlation between stream discharge and effective carry of the formulations, except in the case of Teknar, possibly due to the influence of the physical characteristics of the streams where evaluation was conducted.

J Biochem Biophys Methods, 1990 Mar, 20(3), 237 - 46
Analysis of cyclomaltodextrin glucanotransferase isoenzymes by isoelectric focusing in immobilized pH gradients; Mattsson P et al.; The catalytically active subforms of cyclomaltodextrin glucanotransferase (CGTase; EC 2.4.1.19) from Bacillus circulans var . alkalophilus and from a strain in which the CGTase expressing gene had been cloned were studied by using isoelectric focusing (IEF) in immobilized and in conventional pH gradients . Even with high protein loads the best resolution was achieved in immobilized pH gradients (IPG) . Native CGTase, focused on IPG 4.5-5.4, was resolved into more than 6 subforms, a major one with pI 4.97 and the others between pH 4.75 and 4.99 . Cloned CGTase focused on the same IPG gel was resolved into more than 7 subforms over the pH range 4.78-5.22, with one major component of pI 5.20 . A sensitive assay for the detection of CGTase on analytical gel slices was developed . The different behaviour between native and cloned CGTases was most probably a consequence of the presence--in the latter--of four extra amino acids at the N-terminus . A study of the reaction products by the enzymatic subforms confirmed that they are catalytically identical, hence small differences outside the active site are responsible for the presence microheterogeneity of the protein.

An Acad Bras Cienc, 1990 Mar, 62(1), 85 - 94
Production of a bacteriophage, a phage tail-like bacteriocin and an antibiotic by Bacillus azotofixans; Seldin L et al.; Twenty-two nitrogen-fixing Bacillus azotofixans strains were shown to produce an inhibition zone against themselves in plate assays . The B . azotofixans type strain P3L-5, chosen for further studies, produced inhibition zones against various Bacillus strains and other bacterial genera . This antibacterial substance was also produced in liquid medium and its production was enhanced in semisolid medium (0.4% agar) after 3 to 5 days of incubation . The substance was suggested to be an antibiotic and its preliminary characterization showed resistance to heat (100 degrees C, 15 minutes), to trypsin, pronase, deoxyribonuclease I, ribonuclease A, phospholipase C, ethanol, acetone, and ether, and sensitivity to strong alkali treatment . Its molecular weight was estimated to be between 3500 to 6000 . After induction of B . azotofixans P3L-5 with mitomycin C or ultraviolet light, two types of particles were detected in the lysate: one similar to a phage tail and the other, less frequent, similar to a complete bacteriophage . Lysates containing these particles showed a killing effect in some but not all B . azotofixans strains, but neither the other Bacillus species nor Micrococcus were inhibited by these lysates.

Biull Eksp Biol Med, 1990 Mar, 109(3), 299 - 301
{The genetic transfer, heredity and phenotypic expression of plasmid RP4::Mu cts genes in Bacillus cereus}; Timakova NV et al.; The transcipients were obtained in intrageneric matings of E.coli donor harbouring the plasmid PR4::Mu cts 62 with Bac . cereus GP7 recipient cells with the frequency 10(-9) . The transcipient clone Bac . cereus 682 was selected having stably inherited and expressed the hybrid plasmid PR4::Mu cts 62 genes for antibiotic resistance and temperature sensitivity . Production of the bacteriophage Mu cts 62 particles was not registered in the bacillary transcipient cells . The plasmid RP4::Mu cts 62 genes were localized in the chromosome of Bac . cereus 682 transcipient by the blot-hybridization technique with 32P-labelled DNA of the bacteriophage Mu cts 62 and the plasmid PR4 . The transcipient of Bac . cereus 682 has the donor properties and transfers the RP4::Mu cts 62 genes to recipient cells of Bac . cereus DSM 318 with the frequency of 10(-6)-10(-7) . The expression and transfer of the gram-negative plasmid genes in gram-positive bacterial cells are discussed.

Z Naturforsch {C}, 1990 Mar-Apr, 45(3-4), 240 - 4
Excretion into the culture medium of a Bacillus beta-glucanase after overproduction in Escherichia coli; Riethdorf S et al.; The beta-glucanase gene (bgl) from Bacillus amyloliquefaciens was expressed in E . coli CSH 55 under the control of the PR promoter of phage lambda that is repressed by the thermosensitive repressor C1857 . Production of beta-glucanase was drastically stimulated by a temperature shift to 42 degrees C . This overexpression of the bgl gene (about 20% of the total cellular protein) led to an almost complete excretion of the otherwise periplasmic protein into the extracellular medium, beta-glucanase accounted for more than 50% of the extracellular proteins . Col E 1 related plasmid (pEG 1) are amplified in E . coli relA strains in response to an amino acid limitation leading to a 10-fold increase in the activity of plasmid encoded genes . In this work we intended to maximize the expression of the bgl gene by a concerted action of a plasmid amplification and temperature induction . Surprisingly we could not increase the beta-glucanase production above the level reached by plasmid amplification or temperature induction alone . The reasons for this unexpected result will be discussed . Under all conditions tested the expression of the bgl gene was much lower in the E . coli relA strain NF 162 than in E . coli CSH 55; the low beta-glucanase production was accompanied by a reduced excretion rate of the enzyme.

Ital J Biochem, 1990 Mar-Apr, 39(2), 115 - 20
Enzymatic synthesis of purine 2'-deoxyriboside and its properties as an inhibitor of adenosine deaminases from calf intestinal mucosa and Bacillus cereus; Caffaz S et al.; Enzymatic synthesis of purine 2'-deoxyriboside was obtained by reacting purine with excess 2-deoxy-alpha-D-ribose-1-phosphate in the presence of commercial bovine nucleoside phosphorylase; the product was isolated by semipreparative reverse phase HPLC with an overall 62% yield . Purine 2'-deoxyriboside was shown to behave as a competitive inhibitor of adenosine deaminase from calf intestinal mucosa and Bacillus cereus, with apparent Ki values of 4.5 and 8.5 microM, respectively.

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