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 o