Electrophoresis, 1998 Jun, 19(8-9), 1325 - 31 Natural resistance to intracellular parasites: a study by two-dimensional gel electrophoresis coupled with multivariate analysis; Kovarova H et al.; Natural resistance to Mycobacterium bovis bacillus Calmette-Guerin (BCG) is determined by the Bcg gene (Nramp1), which is exclusively expressed by mature macrophages . The Nramp1 gene is a dominant autosomal gene that has two allelic forms; r confers resistance and s confers susceptibility to infection with intracellular pathogen . Although the wide range of pleiotropic immunological effects of the Nramp1 gene has been described, the exact mechanism of its action remains elusive . In this study we searched for differentially expressed proteins that might provide clues in the studies on Nramp1 gene function . We performed two-dimensional gel electrophoresis of cellular proteins prepared from a B10R macrophage line derived from mice carrying the r allele of the Nramp1 gene, B10S macrophages carrying the s allele, and B10R-Rb macrophages transfected with Nramp1-ribozyme . The classification of protein patterns and selection of distinct proteins characteristic of r or s allele-carrying macrophages was performed using the principal component analysis . We found differential expression of four proteins with the following isoelectric point/molecular weight (pI/Mr) in B10R macrophages compared to B10S and B10R-Rb macrophages: 6.6/25, 7.0/22, 9.1/31.5, and 5.3/8.5 . The protein 7.0/22 has been identified as Mn-superoxide dismutase and the best candidate for protein p6.6/25 seems to be Bcl-2 according to the immunoblot analysis . When the splenic macrophages carrying the r or s allele were analyzed, the changes in relative abundance for proteins 6.6/25 and p7.0/22 were satisfactorily reproduced . Overall, the two identified proteins are important in the regulation of intracellular redox balance and the regulation of apoptosis in macrophages, respectively . Our findings may suggest their possible biological role in the innate immunity against intracellular pathogens. Int J Biochem Cell Biol, 1998 May, 30(5), 579 - 95 Purification and partial characterization of a neutral protease from a virulent strain of Bacillus cereus; Sierecka JK; The factors involved in the pathogenesis of Bacillus cereus (B . cereus) in non-gastrointestinal diseases are poorly investigated . Some researchers suggest that B . cereus proteases may be involved in these illnesses . The aim of this work was to purify and characterize a protease isolated from a virulent strain of B . cereus to explain its assumptive damaging effect . The enzyme was purified in a four-step procedure involving ammonium sulfate fractionation, acetone precipitation, Bio-Gel filtration and column chromatography on DEAE-cellulose (DE-52 cellulose) . The enzyme appeared homogenous using disc electrophoresis . The specific activity of the protease was 72 U/mg of protein . The enzyme was shown to have a relative molecular mass of 29 kDa . The protease was most active at pH 7.0 and 40 degrees C with haemoglobin as the substrate . The enzyme was made completely inactive by ethylenediaminetetraacetic acid (EDTA), beta-mercaptoethanol, dithiothreitol (DTT) and benzamidine (at a concentration of 1 mM) and by diisopropylfluorophosphate (DIPF), L-cysteine, L-histidine, 1,10-phenanthroline (at a concentration of 10 mM) . Divalent cations, especially Ca2+ increased enzyme activity . The enzyme hydrolysed haemoglobin, albumin and casein as the substrates . With haemoglobin and albumin as the substrates Michaelis-Menten kinetics was observed . The obtained Km values were 86 +/- 40 microM (SD, n = 3) and 340 +/- 100 microM (SD, n = 3) for haemoglobin and albumin, respectively . The corresponding Vmax values were 1.26 +/- 0.1 (SD, n = 3) and 0.38 +/- 0.07 (SD, n = 3) mumol of tyrosine liberated per min, per ml, and per mg, while those for casein were not determined . It is concluded that this enzyme is a metal-chelator-sensitive, neutral protease damaging haemoglobin and albumin. Biosci Biotechnol Biochem, 1998 Jun, 62(6), 1103 - 8 Phosphopentomutase of Bacillus stearothermophilus TH6-2: the enzyme and its gene ppm; Hamamoto T et al.; Phosphopentomutase catalyzes the transfer of an intramolecular phosphate on ribose or deoxyribose, and is involved in the salvage pathway of nucleoside synthesis . We identified a sequence 5'-upstream of the genes for the nucleoside phosphorylases of Bacillus stearothermophilus as the phosphopentomutase (ppm) gene . The novel gene corresponded to an open reading frame of 1,179 nucleotides that is translated into a putative 393-amino acid protein with a molecular weight of 43,735 . The gene product, partially purified from ppm-overexpressing Escherichia coli cells, was judged to be a monomer of a 44-kDa polypeptide . The phosphopentomutase was found to catalyze the phosphotransfer on not only ribose or deoxyribose but also arabinose or dideoxyribose. Biosci Biotechnol Biochem, 1998 Jun, 62(6), 1093 - 102 Clustered proline residues around the active-site cleft in thermostable oligo-1,6-glucosidase of Bacillus flavocaldarius KP1228; Kashiwabara S et al.; The gene that coded for a cellular oligo-1,6-glucosidase (dextrin 6-alpha-D-glucanohydrolase, EC 3.2.1.10) in Bacillus flavocaldarius KP1228 (FERM-P9542) cells growing at 51-82 degrees C was expressed in Escherichia coli JM109 . The enzyme had a half-life of 10 min at 89.2 degrees C . Purification of the enzyme and its characterization showed that the enzyme was identical with the native one . Its primary structure of 529 residues with a molecular weight of 61,469 deduced from the gene was 40-42% identical to the sequences of less thermostable oligo-1,6-glucosidases from Bacillus cereus ATCC 7064, Bacillus coagulans ATCC 7050, and Bacillus thermoglucosidasius KP1006 . Sequence analysis showed that the B . flavocaldarius enzyme shared 14 proline residues at the same positions as in the three other enzymes, and that the B . flavocaldarius enzyme had 22 of 33 additional proline residues (cf . 1/5, 5/10, and 9/18 in the respective counterparts) in three long polypeptides constituting the active-site cleft, which connected the third, fourth, and eighth beta-strands to the corresponding third, fourth, and eighth alpha-helices in the (beta/alpha)8-barrel. Appl Environ Microbiol, 1998 Aug, 64(8), 3036 - 41 A novel insecticidal toxin from photorhabdus luminescens, toxin complex a (Tca), and its histopathological effects on the midgut of manduca sexta Blackburn M, Golubeva E, Bowen D, Ffrench-Constant RH. Photorhabdus luminescens is a bacterium which is mutualistic with entomophagous nematodes and which secretes high-molecular-weight toxin complexes following its release into the insect hemocoel upon nematode invasion . Thus, unlike other protein toxins from Bacillus thuringiensis (delta-endotoxins and Vip's), P . luminescens toxin (Pht) normally acts from within the insect hemocoel . Unexpectedly, therefore, the toxin complex has both oral and injectable activities against a wide range of insects . We have recently fractionated the protein toxin and shown it to consist of several native complexes, the most abundant of which we have termed Toxin complex a (Tca) . This complex is highly active against the lepidopteran Manduca sexta . In view of the difference in the normal mode of delivery of P . luminescens toxin and the apparent communality in the histopathological effects of other gut-active toxins from B . thuringiensis, as well as cholesterol oxidase, we were interested in investigating the effects of purified Tca protein on larvae of M . sexta . Here we report that the histopathology of the M . sexta midgut is similar to that for other novel midgut-active toxins . Following oral ingestion of Tca by M . sexta, we observed an acceleration in the blebbing of the midgut epithelium into the gut lumen and eventual lysis of the epithelium . The midgut shows a similar histopathology following injection of Tca into the insect hemocoel . These results not only show that Tca is a highly active oral insecticide but also confirm the similar histopathologies of a range of very different gut-active toxins, despite presumed differences in modes of action and/or delivery . The implications for the mode of action of Tca are discussed. Curr Microbiol, 1998 Sep, 37(3), 195 - 200 Distribution, serological identification, and PCR analysis of Bacillus thuringiensis isolated from soils of Korea; Kim HS et al.; A total, 58 strains of Bacillus thuringiensis were isolated from soils of various regions in Korea . Serological tests showed that B . thuringiensis isolates represented 10 H serotypes, indicating a varied flora of B . thuringiensis . But the H serotypes did not have a significantly uneven distribution, ranging from 1 to 11 isolates . In toxicity tests, 35% of all isolates were toxic to lepidoptera, 20% were toxic to diptera, and 9% were non-toxic isolates . Especially, a large number of lepidopteran/dipteran-active isolates (36%) were found . Forty all lepidopteran-active isolates produced typical rhomboidial inclusions, and the remainder, which belong to dipteran-active and non-toxic isolates, were spherical in shape . In addition, lepidopteran/dipteran-active isolates produced rhomboidal or spherical inclusions . PCR analysis using cryI, II, III, IV, and V gene-specific primers showed that the frequency of the cryIC gene (57%) predominated, followed by the cryIA(b) (45%) and cryIIA genes (34%) . But, the cryIE, cryIF, cryIII, cryIVC and cryV genes were not reactive . Several isolates had unusual PCR products and multiple insecticidal crystal protein genes . PCR results showed varied distribution of the cry-type gene . Seven isolates were selected for evaluation of novel activity according to the following criteria: flagellar serotypes, parasporal inclusion morphology, SDS-PAGE, plasmid DNA patterns, toxicity, and the cry-type gene in PCR analysis . Two isolates, named S333 (H7) and S225 (H7), among them synthesized PCR products of the cryIC gene, but the S333 isolate producing rhomboidal inclusion was toxic to both Plutella xylostella and Culex pipiens, whereas the S225 isolate having toxicity to only C . pipiens produced spherical inclusion. Appl Environ Microbiol, 1998 Aug, 64(8), 2995 - 3003 Phage display of a biologically active Bacillus thuringiensis toxin; Kasman LM et al.; Activated forms of Bacillus thuringiensis insecticidal toxins have consistently been found to form insoluble and inactive precipitates when they are expressed in Escherichia coli . Genetic engineering of these proteins to improve their effectiveness as biological pesticides would be greatly facilitated by the ability to express them in E . coli, since the molecular biology tools available for Bacillus are limited . To this end, we show that activated B . thuringiensis toxin (Cry1Ac) can be expressed in E . coli as a translational fusion with the minor phage coat protein of filamentous phage . Phage particles displaying this fusion protein were viable, infectious, and as lethal as pure toxin on a molar basis when the phage particles were fed to insects susceptible to native Cry1Ac . Enzyme-linked immunosorbent assay and Western blot analysis showed the fusion protein to be antigenically equivalent to native toxin, and micropanning with anti-Cry1Ac antibody was positive for the toxin-expressing phage . Phage display of B . thuringiensis toxins has many advantages over previous expression systems for these proteins and should make it possible to construct large libraries of toxin variants for screening or biopanning. Antimicrob Agents Chemother, 1998 Aug, 42(8), 2055 - 9 vanA gene cluster in a vancomycin-resistant clinical isolate of Bacillus circulans; Ligozzi M et al.; We report on the cloning and sequencing of the vanA gene cluster present in the glycopeptide-resistant clinical isolate Bacillus circulans VR0709 (R . Fontana, M . Ligozzi, C . Pedrotti, E . M . Padovani, and G . Cornaglia, Eur . J . Clin . Microbiol . Infect . Dis . 16:473-474, 1997) . The presence of a vanA-related gene in VR0709 was demonstrated in a PCR assay which permitted the specific amplification of an internal segment of vanA . Southern blotting suggested that the vanA gene was located in the chromosome in a 7 . 6-kb EcoRI fragment . DNA sequence analysis revealed the presence of all seven genes of the vanA cluster (vanR, vanS, vanH, vanA, vanX, vanY, and vanZ) . The degree of identity between homologous proteins encoded by Tn1546 and the chromosome of B . circulans VR0709 ranged from 87 to 95% . Neither PCR nor Southern blotting with specific primers and probes, respectively, showed the presence of open reading frames (ORFs) 1 and 2 which encode the transposase and the resolvase of Tn1546, respectively, the transposon found to carry the vanA gene cluster in enterococci . Determination of the sequences of the flanking regions of the van gene cluster of B . circulans revealed perfect inverted repeats of 10 bp which delineated a 9.2-kb region containing the van gene cluster and an ORF which encoded a putative protein (178 residues) which displayed a low level of identity (28%) to the resolvase of Tn1546 . These results suggest that glycopeptide resistance in B . circulans VR0709 is associated with the acquisition of a vanA gene cluster which shows a high degree of homology with that of enterococci . In B . circulans, however, the cluster is not carried by Tn1546 and is borne by the chromosome. Acta Paediatr, 1998 Jun, 87(6), 702 - 4 Disseminated Bacillus Calmette-Guérin infection in a girl with hyperimmunoglobulin E syndrome; Pasic S et al.; Disseminated Bacillus Calmette-Guerin infection occurs in few well-defined immunodeficiencies, such as severe combined immunodeficiency, chronic granulomatous disease and paediatric acquired immunodeficiency syndrome . This severe complication of immunization against tuberculosis has been lethal in the majority of children who had primary immunodeficiency . Our patient, a 9-y-old girl with hyperimmunoglobulin E syndrome developed disseminated Bacillus Calmette-Guerin infection in infancy . Patients with hyperimmunoglobulin E syndrome (HIES) are susceptible to serious staphylococcal and fungal infections . Disseminated Bacillus Calmette-Guerin infection has not previously been reported in this rare immunodeficiency. J Am Dent Assoc, 1998 Jul, 129(7), 985 - 91 How well does the Chemiclave sterilize handpieces? Kolstad RA. Using the Food and Drug Administration's protocol for testing health care sterilizers, the author investigated the ability of chemical vapor and steam to sterilize handpieces . Five internal sites of six high-speed handpiece models and four internal positions of one low-speed handpiece model were each inoculated with 10(6) Bacillus stearothermophilus spores . Half-cycle challenges were conducted with Chemiclave models EC 5500 and 8000 (Barnstead/Thermolyne) and with two autoclaves, Tuttnauer 2540M (large chamber) (Tuttnauer USA Co., Ltd.) and Statim Cassette (SciCan USA) . Experiments with spores either openly exposed or partially enclosed prove that chemical vapor is an excellent lethal agent, but strongly suggest penetration weakness {corrected}. FEMS Immunol Med Microbiol, 1998 Jun, 21(2), 117 - 22 Effect of ribonuclease from Bacillus intermedius on human blood lymphocytes; Kurinenko BM et al.; By using a rosette formation test the effect of ribonuclease Bacillus intermedius (RNase Bi) on T- and B-lymphocytes in human peripheral blood has been studied in vitro . The RNase effect on T-lymphocytes depends on its concentration: low concentrations (10(-6)-10(-2) microg ml(-1)) stimulate E-rosette formation whereas high concentrations (10 microg ml(-1)) suppress it . The amount of B-lymphocytes decreases under RNase Bi influence in all concentrations tested . RNase Bi like thymus hormones influence immature lymphocytes (0-cells) by inducing the surface expression of E-receptors what leads to rosette formation and, thus, contributes to lymphocyte differentiation . The increase in the amount of active T-cells which represent the mature cell population also confirms the participation of RNase Bi in T-rank lymphocytes differentiation processes . The RNase Bi effect on the human lymphocytes depends on its catalytic activity. Protein Sci, 1998 Jul, 7(7), 1538 - 44 Scan-rate dependence in protein calorimetry: the reversible transitions of Bacillus circulans xylanase and a disulfide-bridge mutant; Davoodi J et al.; The stabilities of Bacillus circulans xylanase and a disulfide-bridge-containing mutant (S100C/N148C) were investigated by differential scanning calorimetry (DSC) and thermal inactivation kinetics . The thermal denaturation of both proteins was found to be irreversible, and the apparent transition temperatures showed a considerable dependence upon scanning rate . In the presence of low (nondenaturing) concentrations of urea, calorimetric transitions were observed for both proteins in the second heating cycle, indicating reversible denaturation occurs under those conditions . However, even for these reversible processes, the DSC curves for the wild-type protein showed a scan-rate dependence that was similar to that in the absence of urea . Calorimetric thermograms for the disulfide mutant were significantly less scan-rate dependent in the presence of urea than in the urea-free buffer . The present data show that, just as for irreversible transitions, the apparent transition temperature for the reversible denaturation of proteins can be scan-rate dependent, confirming the prediction of Lepock et al . (Lepock JR, Rithcie KP, Kolios MC, Rodahl AM, Heinz KA, Kruuf J, 1992, Biochemistry 31:12706-12712) . The kinetic factors responsible for scan-rate dependence may lead to significant distortions and asymmetry of endotherms, especially at higher scanning rates . This points to the need to check for scan-rate dependence, even in the case of reversible denaturation, before any attempt is made to analyze asymmetric DSC curves by standard thermodynamic procedures . Experiments with the disulfide-bridge-containing mutant indicate that the introduction of the disulfide bond provides additional stabilization of xylanase by changing the rate-limiting step on the thermal denaturation pathway. Appl Microbiol Biotechnol, 1998 Jun, 49(6), 737 - 42 Trans activation of the Escherichia coli ato structural genes by a regulatory protein from Bacillus megaterium: potential use in polyhydroxyalkanoate production Pettinari MJ, Vazquez GJ, Kruger N, Vary PS, Steinbuchel A, Mendez BS. A Bacillus megaterium genomic fragment, which encoded an activator homologous to sigma 54 regulators and which was capable of activating Escherichia coli ato genes in trans, was detected in a gene library of B . megaterium screened for beta-ketothiolase activity . The fragment presented only one complete open reading frame (ORF1), which encoded a protein of 398 amino acids . The recombinant plasmid complemented mutations in the Escherichia coli atoC regulatory gene . The constitutive expression of the E . coli ato operon mediated by ORF1 could be useful for the synthesis of polyhydroxyalkanoates with different flexibility properties by recombinant E . coli strains. Vaccine, 1998 Jul, 16(11-12), 1166 - 71 Studies of vaccination of persons in close contact with leprosy patients in Argentina; Bottasso O et al.; A total of 670 adults living or working with leprosy patients, were examined for a BCG vaccination scar, and skin-tested with four new tuberculins . Based on the results 513 were vaccinated, 65 with Bacille de Calmette et Guerin (BCG) alone, 66 with BCG plus killed Mycobacterium vaccae and 382 with killed M . vaccae alone . Skin-testing was repeated 2-3 years later on 344 subjects, when all three vaccines were found to have been highly successful in increasing responses to Tuberculin and Leprosin A (p < 0.0005) with increased immune recognition of common and species-specific antigens . Mean diameters of induration to each skin-test were greatest in recipients of BCG alone (p < 0.05), which suggests that better immuno-regulation occurs after receiving vaccines that incorporate M . vaccae . The results suggest 10(8) M . vaccae alone might prove a valuable future vaccine, which would not require selective pre-vaccination procedures. Eur Cytokine Netw, 1998 Jun, 9(2), 181 - 6 BCG-induced interleukin-6 upregulation and BCG internalization in well and poorly differentiated human bladder cancer cell lines; Bevers RF et al.; Intravesical bacillus Calmette-Guerin (BCG) is a successful therapy for superficial bladder cancer . However, the working mechanism of BCG after intravesical instillation is not completely understood . A functional role of urothelial (tumor) cells in the initiation of the BCG-induced immune reaction should be considered . Here, the possibility of a causal relationship between BCG-induced interleukin-6 (IL-6) synthesis and BCG internalization by urothelial tumor cells was examined in a series of human transitional bladder cancer (TCC) cell lines with different degrees of differentiation . The results showed that the well differentiated TCC cell lines, RT4, SBC-2, and SBC-7, did not possess the capacity to internalize BCG, which was associated with an inability to upregulate IL-6 synthesis when stimulated with BCG . Moreover, these cell lines expressed a low level of constitutive IL-6 synthesis . In contrast, the poorly differentiated TCC cells, T-24, TCC-SUP and J-82, were able to internalize BCG . In T24 and J82, but not in TCC-SUP cells, BCG internalization appeared to result in an upregulation of IL-6 synthesis . Constitutive IL-6 synthesis of the high grade cell lines was found to be cell line-dependent: both TCC-SUP and J82 cells exhibited a high level of constitutive IL-6 synthesis, whereas T24 cells exhibited a low level . The possible relationship between BCG internalization and IL-6 upregulation was studied in detail with the T24 cell line, which exhibited a low constitutive and high BCG-inducible IL-6 synthesis, using anti-BCG antibodies (alphaBCG) and Cytochalasin B as internalization inhibitors . Upregulation of IL-6 synthesis was significantly inhibited by alphaBCG or Cytochalasin B, indicating that internalization is a prerequisite for BCG-induced upregulation of IL-6 synthesis . In conclusion, upregulation of IL-6 production due to BCG internalization by poorly differentiated bladder carcinoma cells may be part of the mode of action of intravesical BCG therapy. Extremophiles, 1997 Aug, 1(3), 151 - 6 Thermostable alkaline cellulase from an alkaliphilic isolate, Bacillus sp . KSM-S237; Hakamada Y et al.; Thermostable alkaline cellulase (endo-1,4-beta-glucanase, EC 3.2.1.4) activity was detected in the culture medium of a strictly alkaliphilic strain of Bacillus, designated KSM-S237 . This novel enzyme was purified to homogeneity by a two-step column-chromatographic procedure with high yield . The N-terminal amino acid sequence of the purified enzyme was Glu-Gly-Asn-Thr-Arg-Glu-Asp-Asn-Phe-Lys-His-Leu-Leu-Gly-Asn-Asp-Asn-Val- Lys-Arg . The enzyme had a molecular mass of approximately 86 kDa and an isoelectric point of pH 3.8 . The enzyme had a pH optimum of 8.6-9.0 and displayed maximum activity at 45 degrees C . The alkaline enzyme was stable up to 50 degrees C and more than 30% of the original activity was detectable after heating at 100 degrees C and at pH 9.0 for 10 min . The enzyme hydrolyzed carboxymethylcellulose, lichenan (beta-1,3;1,4-linkage), and p-nitrophenyl derivatives of cellotriose and cellotetraose . Crystalline forms of cellulose (Avicel and filter paper), H3PO4-swollen cellulose, NaOH-swollen cellulose, curdlan (beta-1,3-linkage), laminarin (beta-1,3;1,6-linkage), and xylan were barely hydrolyzed at all. Extremophiles, 1997 Aug, 1(3), 135 - 41 K+/H+ antiporter in alkaliphilic Bacillus sp . no . 66 (JCM 9763); Kitada M et al.; A K+/H+ antiport system was detected for the first time in right-side-out membrane vesicles prepared from alkaliphilic Bacillus sp . no . 66 (JCM 9763) . An outwardly directed K+ gradient (intravesicular K+ concentration, K(in), 100 mM; extravesicular K+ concentration, K(out), 0.25 mM) stimulated uphill H+ influx into right-side-out vesicles and created the inside-acidic pH gradient (delta pH) . This H+ influx was pH-dependent and increased as the pH increased from 6.8 to 8.4 . Addition of 100 microM quinine inhibited the H+ influx by 75% . This exchange process was electroneutral, and the H+ influx was not stimulated by the imposition of the membrane potential (interior negative) . Addition of K+ at the point of maximum delta pH caused a rapid K+-dependent H+ efflux consistent with the inward exchange of external K+ for internal H+ by a K+/H+ antiporter . Rb+ and Cs+ could replace K+ but Na+ and Li+ could not . The H+ efflux rate was a hyperbolic function of K+ and increased with increasing extravesicular pH (pH(out)) from 7.5 to 8.5 . These findings were consistent with the presence of K+/H+ antiport activity in these membrane vesicles. Extremophiles, 1997 May, 1(2), 61 - 6 Alkaline cellulases from alkaliphilic Bacillus: enzymatic properties, genetics, and application to detergents; Ito S; We have isolated a number of alkaliphilic Bacillus that produce alkaline exoenzymes and found a possible use for alkaline cellulase (carboxymethylcellulase) as an additive for improving the cleaning effect of detergents . Enzymatic properties of some candidate cellulases fulfilled the essential requirements for enzymes to be used practically in laundry detergents . Here I describe the properties and possible catalytic mechanism of the hydrolytic reaction and the gene for the industrial alkaline cellulase produced by one of the isolates, Bacillus sp . KSM-635. Extremophiles, 1997 Nov, 1(4), 199 - 206 Isolation and characterization of bacteriophage BCJA1, a novel temperate bacteriophage active against the alkaliphilic bacterium, Bacillus clarkii; Jarrell KF et al.; The isolation and characterization of a novel bacteriophage active against the obligately alkaliphilic bacterium Bacillus clarkii is described . The bacteriophage, designated BCJA1 . is a member of the Siphoviridae family with a B1 morphology . It possesses an isometric head, which measures 65 nm between opposite apices, and a noncontractile tail of 195 nm length . It had a buoyant density of 1.518 g/ml and an estimated particle mass of 37 x 10(7) daltons . BCJA1 was stable over the pH range of 6-11 . A one-step growth experiment conducted at pH 10 demonstrated a latent period of about 40 min and a burst size of approximately 40 . The purified bacteriophage appeared to consist of 10 proteins with the major head and tail proteins likely to be of molecular weight 36500 and 28000, respectively . The genome size was estimated to be between 32.1 and 34.8 kb . The percent G + C content of purified bacteriophage DNA was 45.6 . The wildtype bacteriophage is temperate but a clear plaque mutant was isolated. Extremophiles, 1997 Nov, 1(4), 163 - 9 Mechanisms of cytoplasmic pH regulation in alkaliphilic strains of Bacillus; Krulwich TA et al.; The central challenge for extremely alkaliphilic Bacillus species is the need to establish and sustain a cytoplasmic pH that is over two units lower than the highly alkaline medium . Its centrality is suggested by the strong correlation between the growth rate in the upper range of pH for growth, i.e., at values above pH 10.5, and the cytoplasmic pH . The diminishing growth rate at extremely high pH values correlates better with the rise in cytoplasmic pH than with other energetic parameters . There are also general adaptations of alkaliphiles that are crucial prerequisites for pH homeostasis as well as other cell functions, i.e., the reduced basic amino acid content of proteins or segments thereof that are exposed to the medium, and there are other challenges of alkaliphily that emerge from solution of the cytoplasmic pH problem, i.e., reduction of the chemiosmotic driving force . For cells growing on glucose, strong evidence exists for the importance of acidic cell wall components, teichuronic acid and teichuronopeptides, in alkaliphily . These wall macromolecules may provide a passive barrier to ion flux . For cells growing on fermentable carbon sources, this and other passive mechanisms may have a particularly substantial role, but for cells growing on both fermentable and nonfermentable substrates, an active Na+-dependent cycle is apparently required for alkaliphily and the alkaliphile's remarkable capacity for pH homeostasis . The active cycle involves primary establishment of an electrochemical gradient via proton extrusion, a secondary electrogenic Na+/H+ antiport to achieve net acidification of the cytoplasm relative to the outside pH, and mechanisms for Na+ re-entry . Recent work in several laboratories on the critical antiporters involved in this cycle has begun to clarify the number and characteristics of the porters that support active mechanisms of pH homeostasis. J Urol, 1998 Aug, 160(2), 394 - 6 Treatment of recurrent penile condylomata acuminata with external application and intraurethral instillation of bacillus Calmette-Guerin; Bohle A et al.; PURPOSE: Condylomata acuminata are caused by human papillomavirus infection . Despite numerous treatment modalities these patients often demonstrate recurrent disease . We report initial experience with bacillus Calmette-Guerin (BCG) therapy in patients not responding to standard treatment . MATERIALS AND METHODS: Between October 1994 and March 1997, 6 men with rapidly recurrent external and intraurethral condylomata acuminata underwent BCG therapy after initial laser treatment . External application and intraurethral instillation of BCG were performed 6 times in weekly intervals . Followup studies included examination and endoscopic inspection of the urethra and bladder . RESULTS: Of the patients 3 completed 1 course of BCG and had no relapse of condylomata acuminata, 2 underwent a second course of BCG and 1 had relapse, and 1 had relapse after discontinuing therapy due to penile edema . The annual recurrence rate decreased from 3.2 before to 0.75 after BCG therapy (p < 0.05, test of equality of 2 percentages) . CONCLUSIONS: Immunotherapy with BCG is accepted treatment for superficial transitional cell carcinoma . The BCG induced immune response appears to reduce the recurrence rate in patients with condylomata acuminata. Biochem Mol Biol Int, 1998 Jul, 45(3), 443 - 52 Production and purification of an extracellular cellulase from Bacillus brevis VS-1; Singh VK et al.; A bacteria identified as Bacillus brevis has been isolated from the soil . It has been found to secrete cellulase extracellularly whose production increased almost five times on addition of galactose in the culture medium . Production of cellulase has been found optimal at pH 5.5, 37 degrees C and 175 rpm speed using environmental orbital shaker . The cellulase has been purified using ultrafiltration and Sephadex G-200 column chromatography . The native molecular weight of the enzyme is found to be 33,000 +/- 2000 using Sephadex G-200 gel filtration chromatography . The subunit molecular weight (33,000 +/- 2,000) indicate monomeric nature of the enzyme . The enzyme showed Michaelis Menten kinetics exhibiting Km approximately 1.7 +/- 0.1 mg/ml for CMC . The enzyme activity got inhibited by heavy metals viz . Hg+2 and Ag+2. J Biomol NMR, 1998 Feb, 11(2), 185 - 90 High-resolution detection of five frequencies in a single 3D spectrum: HNHCACO--a bidirectional coherence transfer experiment; Pang Y et al.; A new triple-resonance pulse sequence, 3D HNHCACO, is introduced and discussed, which identifies sequential correlations of the backbone nuclei (H alpha (i-1), C alpha (i-1), C(i-1), NH(i), N(i)) of doubly labeled proteins in H2O . The three-dimensional (3D) method utilizes a recording of 15N and 13C resonances in a single indirect time domain, the 13C' resonance in another indirect time domain, and detects both NH and H alpha protons . A bidirectional coherence transfer (NH(i) <--> N(i) <--> C(i-1) <--> C alpha (i-1) <--> H alpha (i-1)) is effectuated, resulting in a single high-resolution 3D spectrum that contains the frequencies of all five backbone nuclei . The experiment was applied to the 12.3 kDa ribonuclease from Bacillus intermedius (Binase). J Food Prot, 1998 Jul, 61(7), 921 - 3 Identification of Bacillus cereus by Fourier transform infrared spectroscopy (FTIR); Lin SF et al.; The objective of this study was to evaluate the potential of Fourier transform infrared spectroscopy (FTIR) for rapid identification of Bacillus cereus isolates . Ten B . cereus group isolates (comprising B . cereus, Bacillus mycoides, and Bacillus thuringiensis strains), five other Bacillus spp., and five non-Bacillus spp . were used . Two types of media, brain heart infusion (BHI) and Trypticase soy agar (TSA), were tested . The results indicated that all B . cereus group isolates produced characteristic absorbance peaks at wave numbers between 1738 and 1740 cm-1 . These peaks were not affected by the growth medium . None of the other bacteria tested showed a similar peak after growth on BHI or TSA . Absorbance peaks between 1800 and 1500 cm-1 of members of the B . cereus group had different shapes and sizes, suggesting that FTIR may be useful for rapid identification of species within the B . cereus group. Biochem J, 1998 Aug 1, 333 ( Pt 3), 677 - 83 Bacillus thuringiensis Cry1Ac toxin interaction with Manduca sexta aminopeptidase N in a model membrane environment; Cooper MA et al.; The Bacillus thuringiensis Cry1Ac delta-endotoxin was shown to bind in a biphasic manner to Manduca sexta aminopeptidase N (APN) present in a novel model membrane . Surface plasmon resonance analysis allowed the quantification of toxin binding to M . sexta APN in a supported lipid monolayer . The initial binding was rapid and reversible, with an affinity constant of 110 nM . The second phase was slower and resulted in an overall affinity constant of 3.0 nM . Reagents used to disrupt protein-protein interactions did not dissociate the toxin after high-affinity binding was attained . The initial association between Cry1Ac and APN was inhibited by the sugar GalNAc, but the higher-affinity state was resistant to GalNAc-induced dissociation . The results suggest that after binding to M . sexta APN, the Cry1Ac toxin undergoes a rate-limiting step leading to a high-affinity state . A site-directed Cry1Ac mutant, N135Q, exhibited a similar initial binding affinity for APN but did not show the second slower phase . This inability to form an irreversible association with the APN-lipid monolayer helps explain the lack of toxicity of this protein towards M . sexta larvae and its deficient membrane-permeabilizing activity on M . sexta midgut brush border membrane vesicles. J Nat Prod, 1998 Jul, 61(7), 939 - 41 Gibbilimbols A-D, cytotoxic and antibacterial alkenylphenols from Piper gibbilimbum; Orjala J et al.; Fractionation of the petroleum ether extract from the leaves of Piper gibbilimbum collected in Papua New Guinea afforded four new alkenylphenols, gibbilimbols A-D (1-4) . The structures of the isolates were elucidated by spectroscopic methods, mainly 1D- and 2D-NMR spectroscopy . Gibbilimbols A-D were found to be toxic to brine shrimp with an LC50 of approximately 5 microg/mL . Gibbilimbols A-D were further found to be cytotoxic toward KB nasopharyngal carcinoma cells (ED50 7.8-2.1 microg/mL) . All isolates also showed antibacterial activity toward Staphylococcus epidermidis and Bacillus cereus. Orv Hetil, 1998 Jun 28, 139(26), 1563 - 70 {Results of the BCG vaccination in Hungary since 1929: evaluation of preventive and immunotherapeutic effectiveness}; Lugosi L; BACKGROUND: The BCG (Bacille Calmette-Guerin), a living attenuated bacterial vaccine with a characteristic residual virulence, has been used to prevent tuberculosis since 1921 (in Hungary non-systematically since 1929) and applied for immunostimulation in neoplasia since the 1960s . MEASURES: Considering the grave tuberculosis epidemiological situation in Hungary, the BCG revaccination became compulsory up to 20 years old tuberculin negatives since 1959 . The Pasteur P1173P2 BCG strain has been used for vaccine manufacturing with improved quality control methods according to the requirements of the WHO . With in systematic BCG primo and revaccination policy 8.1 million BCG vaccination from 1959 to 1983 then further 3.1 million between 1984 and 1996 have been performed . RESULTS: Linear regression analysis demonstrates that the decrease of the TB incidence in children was 3-5 times more rapid (annual average decrease was 25.5%) than in adult since 1959 . Multiple regression analysis indicates that the BCG is the strongest explanatory variable decreasing children TB incidence among other antituberculosis measures . The BCG vaccination efficacy ins demonstrated by 2 x 2 table analysis . The systematic BCG vaccination, the living and persisting BCG in the macrophages, confers acquired resistance against virulent TB infections . The immunostimulation in neoplasia has been applied with concentrated BCG developed in Hungary since 1979 . The adverse reactions are at accepted frequency . The number of BCG vaccinated subjects was estimated at 1.5 billion from 1948 to 1974 in the world . The yearly number of BCG vaccination in the WHOI-EPI System is estimated 50-100 million . CONCLUSION: The efficacy of the BCG vaccination can only be ensured if the vaccine is manufactured and controlled with standardized methods, and applied in a systematic vaccination programme . The effectiveness has to be evaluated in statistically valid biostatistical models. FEMS Microbiol Lett, 1998 Jul 1, 164(1), 201 - 6 Discrimination among Bacillus cereus, B . mycoides and B . thuringiensis and some other species of the genus Bacillus by Fourier transform infrared spectroscopy; Beattie SH et al.; Fourier transform infrared spectroscopy (FTIR) in conjunction with canonical variate analysis was found to be effective in discriminating among spectra of 9 representative strains of Bacillus spp., including B . cereus, B . mycoides and B . thuringiensis . The method was also able to discriminate according to species among spectra of 14 other non-type strains of B . cereus, 12 of B . mycoides and 12 of B . thuringiensis with a success rate of > 95%, even without using a prior classification of the groups by species . FTIR spectroscopy can be used for the rapid and accurate differentiation of species in the genus Bacillus that are of importance to the food and dairy industry. Curr Opin Pulm Med, 1998 May, 4(3), 154 - 61 Examining the biology of tuberculosis; Gonzalez-Rothi RJ; The task of critically reviewing current advances in a scientifically burgeoning topic such as tuberculosis is daunting . From 2965 published articles on tuberculosis in 1997 cited in MEDLINE, 180 were selected, and approximately one third of those are included in this review . This paper highlights how molecular biology has greatly advanced our understanding of the epidemiology and transmission of tuberculosis worldwide . In addition, recent data on the dynamics of transmission between close contacts are provided that may challenge conventional views . A global view of drug-resistant tuberculosis is presented, and new data on the prevalence of tuberculosis in diabetics, bone marrow transplant recipients, patients with renal disease, and HIV-infected persons are highlighted . Recent studies on the genetic susceptibility of tuberculosis are outlined, and we review immunotherapy and the impact of tuberculosis on the natural course of HIV infection . Chemoprevention, duration of therapy, and bacille Calmette-Guerin vaccination in the setting of HIV are discussed . The value and limitation of nucleic acid amplification tests in rapid diagnosis is addressed, and old diagnostic tests are revisited with new data . Novel mycobacterial culture systems are discussed and screening-for-infection approaches are revisited, including tuberculin skin testing in bacille Calmette-Guerin vaccinees . Therapeutic issues addressing antituberculous drug absorption in HIV-infected patients, the role of pharmacokinetics in predicting clinical outcomes, and important drug-drug interactions are discussed. Biochim Biophys Acta, 1998 Jul 28, 1386(1), 199 - 210 Catalytic activity of thermolysin under extremes of pressure and temperature: modulation by metal ions; Kudryashova EV et al.; The catalytic activity of thermolysin (TL), a Zn-dependent neutral protease from Bacillus thermoproteolyticus, has been studied over a wide interval of pressures (1 bar to 4 kbar) and temperatures (20 degreesC to 80 degreesC) by monitoring hydrolysis of a low-molecular-mass substrate, 3-(2-furylacryloyl)-glycyl-L-leucine amide . This reaction shows a very large negative value for the activation volume and, because of that, simultaneous increase in temperature and pressure leads to a significant (up to 40-fold) acceleration of the reaction . At pressures higher than 2-2.5 kbar, the reaction rate starts to decrease due to disactivation of TL . This disactivation is explained in part by pressure-promoted dissociation of zinc ion from the active site and can be inhibited by adding exogenous zinc . Thus, this thermostable protease does not specifically show a higher stability at high pressure in comparison with small mesophilic proteases. Biophys J, 1998 Aug, 75(2), 1010 - 5 Obfuscation of allosteric structure-function relationships by enthalpy-entropy compensation; Tlapak-Simmons VL et al.; The pH and temperature dependence of the allosteric properties of phosphofructokinase (PFK) from Bacillus stearothermophilus have been studied from 5 to 9 and 6 to 40 degrees C, respectively . Throughout this pH and temperature range the allosteric ligands MgADP and phospho(enol)pyruvate (PEP) have no effect on kcat . The dissociation constants of the substrate, fructose 6-phosphate, and the allosteric ligands, as well as the absolute value of the coupling free energies between these ligands, all increase when the pH is raised, indicating that the inhibition by PEP and the activation by MgADP increase despite each ligand's somewhat lower affinity . However, the constituent coupling enthalpies and entropies substantially diminish in absolute value as pH is increased, suggesting that the magnitudes of molecular perturbations engendered by the binding of allosteric ligands do not correlate with the magnitudes of the functional consequences of those perturbations . Temperature and pH exert their influence on the observed allosteric behavior by changing the relative contributions made by the largely compensating DeltaH and TDeltaS terms to the coupling free energy. Biochem Biophys Res Commun, 1998 Jul 20, 248(2), 372 - 7 Improved thermostability of a Bacillus alpha-amylase by deletion of an arginine-glycine residue is caused by enhanced calcium binding; Igarashi K et al.; alpha-Amylase from alkaliphilic Bacillus KSM-1378 (LAMY) is a novel semi-alkaline enzyme which has a high specific activity, a value 5-fold higher than that of a Bacillus licheniformis enzyme at alkaline pH . Thermostability of this enzyme could be improved by deletion of the Arg181-Gly182 residue by means of site-directed mutagenesis . The wild-type and engineered LAMYs were very similar with respect to specific activity, pH-activity curve, temperature-activity curve, susceptibility to inhibitors, and pattern of hydrolysis products from soluble starch and maltooligosaccharides . However, the engineered enzyme also acquired increased pH stability and resistance to sodium dodecyl sulfate and especially chelating reagents, such as ethylenediaminetetraacetate and ethyleneglycol-bis (beta-aminoethylether)tetraacetate . This is the first report that thermostability of alpha-amylase is improved by enhanced calcium binding to the enzyme molecule . Medicina (B Aires), 1997, 57(5), 581 - 6 Cross-reactivity of anti-10kD heat shock protein antibodies in leprosy and tuberculosis patients; Rojas RE et al.; The response to recombinant 10-kD heat shock protein (HSP) of Mycobacterium leprae (rML10) was evaluated by indirect ELISA in sera from leprosy patients, household contacts, tuberculosis patients and healthy controls in a leprosy-endemic area in the North East of Argentina . Some technical parameters were analyzed: within-assay and between-assay variability, dose-response curves and detectability indexes (specificity and sensitivity) of ELISA applied to measure anti-10 kDa antibodies . High levels of these antibodies have already been reported in positive bacilloscopy patients; herein we have also demonstrated that tuberculosis patients sera cross-react with this M . leprae antigen . This test seems to have a low sensitivity and specificity for leprosy detection; it confirms that antibodies against highly conserved HSP antigens are important in the polyclonal response against mycobacterial epitopes in leprosy as well as in tuberculosis. Lett Appl Microbiol, 1998 May, 26(5), 387 - 90 Isolation of a strain of Bacillus thuringiensis ssp . kurstaki HD-1 encoding delta-endotoxin Cry1E; Chang JH et al.; A strain of Bacillus thuringiensis, STB-1, toxic against Spodoptera exigua, was isolated . Bacillus thuringiensis STB-1 produced bipyramidal inclusions and reacted with the H antiserum of B . thuringiensis ssp . kurstaki . The plasmid and protein profiles of B . thuringiensis STB-1 were compared with those of its reference strains, ssp . kurstaki and ssp . kenyae . To verify the gene type of B . thuringiensis STB-1, PCR analysis was performed with Spodoptera-specific cry gene primers . The result showed that B . thuringiensis STB-1, unlike its reference strains, had crylAa, crylAb, crylAc and crylE, suggesting that B . thuringiensis STB-1 was a unique strain with respect to gene type . In addition, B . thuringiensis STB-1 showed a high level of toxicity against both S . exigua and Bombyx mori, whereas B . thuringiensis ssp . kurstaki HD-1 or ssp . kenyae showed a high level of toxicity against only Bombyx mori or S . exigua, respectively. J Appl Microbiol, 1998 May, 84(5), 883 - 8 Characterization of mosquito larvicidal parasporal inclusions of a Bacillus thuringiensis serovar higo strain; Saitoh H et al.; The parasporal inclusion proteins of the type strain of Bacillus thuringiensis serovar higo (H44), that have moderate mosquitocidal activity, were characterized . The purified parasporal inclusions, spherical in shape, were examined for activity against the two mosquito species, Culex pipiens molestus and Anopheles stephensi and the moth-fly, Telmatoscopus albipunctatus . The LC50 values of the inclusion for the two mosquitoes were 3.41 and 0.15 microgram.ml-1, respectively . No mortality was shown for T . albipunctatus larvae by the inclusions at concentrations up to 1 mg ml-1 . Solubilized parasporal inclusions exhibited no haemolytic activity against sheep erythrocytes . Parasporal inclusions consisted of eight proteins with molecular masses of 98, 91, 71, 63, 59, 50, 44 and 27 kDa . Of these, the 50 and 44 kDa proteins were the major components . Analysis with immunoblotting revealed that, among several inclusion proteins of B . thuringiensis serovar israelensis, only two proteins of 130 kDa and 110 kDa reacted weakly with antibodies against higo proteins . N-terminal amino acid sequences of the 98, 91, and 71 kDa proteins showed 85-100% identity to those of the two established Cry protein classes, Cry4A and Cry10A. J Appl Microbiol, 1998 May, 84(5), 791 - 801 Bacillus isolates from the spermosphere of peas and dwarf French beans with antifungal activity against Botrytis cinerea and Pythium species; Walker R et al.; A range of isolation procedures including washing, sonication and incubation in nutrient broth were used separately and in combination to obtain potential bacterial antagonists to Botrytis cinerea and Pythium mamillatum from the testae and cotyledons of peas and dwarf French beans . Heat treatment was also used to bias this selection towards spore-forming bacteria . Ninety-two bacterial isolates were obtained, 72 of which were provisionally characterized as species of Bacillus . Four of these Bacillus isolates (B3, C1, D4 and J7) displayed distinct antagonism in vitro against Botrytis cinerea and P . mamillatum when screened using dual culture analysis . Further characterization of these antagonists using API 50CHB biochemical profiling identified isolate D4 as Bacillus polymyxa and isolates B3, C1 and J7 as strains of B . subtilis . In vitro screening techniques, using cell-free and heat-killed extracts of liquid cultures against Botrytis cinerea, demonstrated the production of antifungal compounds by these four Bacillus antagonists . With each isolate the antifungal activity was found not to be either exclusively spore-bound nor released entirely into the medium but present in both fractions . The antifungal compounds produced by these isolates were shown to be heat-stable . Their identification, production and release require further study for exploitation as biocontrol systems. J Am Mosq Control Assoc, 1998 Jun, 14(2), 183 - 5 Laboratory and field evaluation of efficacy of VectoBac 12AS against Culex sitiens (Diptera: Culicidae) larvae; Brown MD et al.; Laboratory bioassay studies of the efficacy of VectoBac 12AS (active ingredient: 1,200 International Toxic Units {ITU}/mg Bacillus thuringiensis var . israelensis) against field-collected late 3rd/early 4th-instar larvae of Culex sitiens indicated excellent control potential . A 95% lethal concentration (LC95) value of 1.381 x 10(7) ITU was calculated, which equated to a dosage of 0.011 liters/ha . This dosage represented 1.8% of the recommended lowest dosage rate for the product . A field trial of VectoBac 12AS against late 3rd/early 4th-instar field specimens of Cx . sitiens in floating mesh cylinders was then conducted in salt-marsh pools near Coomera Marina, southeast Queensland, Australia . At a rate of 0.5 liters/ha, 100% mortality of Cx . sitiens larvae was recorded at 24 h posttreatment. Infect Immun, 1998 Aug, 66(8), 3643 - 8 An epitope delivery system for use with recombinant mycobacteria; Hetzel C et al.; We have developed a novel epitope delivery system based on the insertion of peptides within a permissive loop of a bacterial superoxide dismutase molecule . This system allowed high-level expression of heterologous peptides in two mycobacterial vaccine strains, Mycobacterium bovis bacille Calmette-Guerin (BCG) and Mycobacterium vaccae . The broader application of the system was analyzed by preparation of constructs containing peptide epitopes from a range of infectious agents and allergens . We report detailed characterization of the immunogenicity of one such construct, in which an epitope from the Der p1 house dust mite allergen was expressed in M . vaccae . The construct was able to stimulate T-cell hybridomas specific for Der p1, and it induced peptide-specific gamma interferon responses when used to immunize naive mice . This novel expression system demonstrates new possibilities for the use of mycobacteria as vaccine delivery vehicles. Rays, 1998 Jan-Mar, 23(1), 225 - 30 BCG and prospects for new vaccines against tuberculosis; Ortona L et al.; Seventy-five years have elapsed since its introduction and a renewed interest has arisen in the vaccination with bacillus Calmette-Guerin (BCG) for the prevention of tuberculosis . This interest has been motivated by the increase in tuberculosis, especially in multidrug-resistant tuberculosis . The efficacy of BCG has been questioned for decades, however, new epidemiological studies have shown a protective effect in some populations and categories at risk . Protection is more evident in the populations with a high incidence of the disease, especially against disseminated and invasive disease . The use of this vaccination is advised for specific populations based on the risk of infection and disease . However, BCG has a limited benefit . New agents produced with methods of molecular biology are supplying encouraging results in the animal model. Rays, 1998 Jan-Mar, 23(1), 9 - 14 General epidemiology of tuberculosis; Ortona L et al.; An outline of the history of tuberculosis is offered for consideration including the present estimated incidence world-wide . The epidemiology of the disease with respect to its etiology is described focusing on the risk of infection, the development of clinical manifestations subsequent to the exposure to the bacillus, and the risk of reactivation . The situation of the epidemiology of tuberculosis in Italy is analyzed based on available information after the closure of TB dispensaries following the introduction of National Health Service in 1978 . In last years over 5000 cases of tuberculosis per year have been notified; however what percentage of the real incidence of TB this data represents, is unknown. FEMS Microbiol Lett, 1998 Jun 15, 163(2), 229 - 36 16S-23S rRNA internal transcribed spacers as molecular markers for the species of the 16S rRNA group I of the genus Bacillus; Daffonchio D et al.; The internal transcribed spacers between the 16S and the 23S ribosomal RNA genes were used to discriminate species of the 16S rRNA group I of the genus Bacillus by PCR . The spacer-PCR fingerprints clearly discriminated the different species, except those closely related like the members of the 'B . cereus group' (B . cereus, B . thuringiensis and B . mycoides) and the species of the 'B . subtilis group' (B . amyloliquefaciens and B . licheniformis) . Examining in more detail the shortest internal transcribed spacers, B . subtilis group species were distinguished by single-strand conformation polymorphism analysis, whereas B . mycoides was differentiated from B . cereus/B . thuringiensis by restriction analysis. Extremophiles, 1998 May, 2(2), 83 - 92 Studies on the respiratory system in alkaliphilic Bacillus; a proposed new respiratory mechanism; Higashibata A et al.; Respiratory electron transfer systems in two alkaliphilic Bacillus species, YN-1 and YN-2000, were investigated In the cyanide-sensitive pathway of the obligate alkaliphilic Bacillus YN-1, the terminal enzyme was a caa3-type cytochrome c oxidase constituting up to just 10% of the total oxygen-reducing activity, while 90% of the respiratory activity was due to cyanide-insensitive, nonproteinaceous material with a molecular weight of 662 . These results were consistent with the cyanide-tolerant growth of the bacterium . The molecular and catalytic properties of the nonproteinaceous material were not identical with those of menaquinones extracted from the bacterium . Furthermore, the nonproteinaceous material was also found in the facultative alkaliphilic Bacillus YN-2000, when that bacterium was cultivated in alkaline conditions . A new respiratory oxygen-reducing mechanism comprising a nonproteinaceous component and a catalase is proposed for these alkaliphilic Bacillus species. Arch Virol, 1997, 142(9), 1933 - 6 Oleavirus, a new genus in the family Bromoviridae; Martelli GP et al.; Oleavirus is a monotypic genus having olive latent virus 2 (OLV-2) as the type species . OLV-2 is transmitted by inoculation of sap but not by aphids . Virus particles have different shape and size, ranging from quasi spherical to bacilliform with length of 37, 43, 48, and 55 nm, respectively, and a diameter of ca . 18 nm . Virions do not contain lipids or carbohydrates and possess a single coat protein species with molecular mass of ca . 24 kDa, which is not required for infectivity . Individual particles contain a single molecule of linear, positive sense ssRNA, constituting ca . 19% of their weight . The genome consists of three functional non polyadenylated, capped, positive sense, single-stranded RNA molecules occurring as three functional species of 3126 nt (RNA1, monocistronic), 2734 nt (RNA2, monocistronic), and 2438 nt (RNA3, bicistronic) . Virions encapsidate a fourth RNA species 2078 nt in size (RNA4) with no apparent messenger activity . Virus replication is thought to occur in the cytoplasm possibly in connection with vesicular structures . The strategy of replication encompasses proteolytic processing and subgenomic RNA production . Oleavirus does not have a complete straightforward relationship with any of the current genera in the Bromoviridae, but shows homologies in diverging directions with one genus of the family or another. Am J Gastroenterol, 1998 Jul, 93(7), 1055 - 9 The immunohistological diagnosis of E . coli O157:H7 colitis: possible association with colonic ischemia; Su C et al.; OBJECTIVE: E . coli O157:H7 may cause hemorrhagic colitis resembling ischemic colitis . Diagnosis is usually made by finding sorbitol-negative colonies on MacConkey agar that react with O157 and H7 antisera . Most ischemic colitis is idiopathic, but some may be caused by E . coli O157:H7, inasmuch as this organism can produce fibrin thrombi in colon vasculature . The objectives of this study were to determine whether E . coli O157:H7 infection can be diagnosed retrospectively from paraffin blocks of colon sections and whether an association exists between E . coli O157:H7 infection and colonic ischemia . METHODS: Paraffin-embedded sections of normal colon (n = 2) and various colitides {ischemic (n = 11), E . coli O157:H7 (n = 2), IBD (n = 8) and pseudomembranous (n = 3)} were used . Sections were deparaffinized, rehydrated, incubated with 3% peroxide in methanol, rinsed, and incubated with peroxidase-labeled antibody isolated from goats immunized with whole E . coli O157:H7 . Sections were stained with peroxidase chromagen reagent and counterstained with hematoxylin . Coarse, granular, orange-brown staining was considered positive . To determine the localization of the chromagen deposits, three cases that stained positive, including one of the culture-proved E . coli O157:H7 colitis and two of colonic ischemia, were processed for electron microscopy . RESULTS: Both cases (100%) of E . coli O157:H7 colitis and three of 11 (27.3%) cases of ischemic colitis stained positive by light microscopy . In one culture-proved case, electron microscopy demonstrated staining of bacillary structures; in two cases of colonic ischemia, extensive deposits of chromagen material were present that were associated neither with inflammatory cells nor with bacterial forms . CONCLUSIONS: Immunoperoxidase staining of archival sections may be used to diagnose E . coli O157:H7 infection . An etiological role for this organism is possible in some cases of colonic ischemia. J Immunol, 1998 Jul 15, 161(2), 1045 - 54 Bacille Calmette-Guérin vaccination enhances human gamma delta T cell responsiveness to mycobacteria suggestive of a memory-like phenotype; Hoft DF et al.; Bacille Calmette-Guerin (BCG) immunity can be studied as one experimental model for mycobacterial protective immunity . We have used flow cytometry to investigate human T cell subsets induced by BCG vaccination . PBMC harvested from BCG-vaccinated individuals and controls were stimulated with mycobacterial Ags, and the T cell subsets present after 7 days of in vitro expansion were characterized . The most dramatic expansions induced by mycobacterial Ags were detected in gamma delta T cells . The gamma delta T cell expansions measured after in vitro stimulation with mycobacterial Ags were significantly greater in BCG responders compared with nonsensitized controls, indicating that BCG vaccination induced gamma delta T cell activation associated with enhanced secondary responses . The majority of gamma delta T cells induced by BCG vaccination were gamma 9+ delta 2+ T cells reactive with isoprenyl pyrophosphates . Coculture with CD4+ T cells induced optimal gamma delta T cell expansion, although IL-2 alone could provide this helper function in the absence of CD4+ T cells . Gamma delta T cells were found to provide helper functions for mycobacterial specific CD4+ and CD8+ T cells as well, demonstrating reciprocal stimulatory interactions between gamma delta T cells and other T cell subsets . Finally, prominent mycobacterial specific gamma delta T cell expansions were detected in a subset of unvaccinated controls with evidence for prior sensitization to mycobacterial lysates (elevated mycobacterial specific lymphoproliferative responses) . These latter findings are consistent with the hypothesis that exposure to atypical mycobacteria or related environmental Ags may induce gamma delta T cells cross-reactive with Ags present in the Mycobacterium tuberculosis complex . Our results suggest that gamma delta T cells may be capable of developing a memory immune-like phenotype, and therefore might be important targets for new vaccines. Microbios, 1998, 93(374), 43 - 54 Comparative antibacterial and antifungal effects of some phenolic compounds; Aziz NH et al.; The antimicrobial potential of eight phenolic compounds isolated from olive cake was tested against the growth of Escherichia coli, Klebsiella pneumoniae, Bacillus cereus, Aspergillus flavus and Aspergillus parasiticus . The phenolic compounds included p-hydroxy benzoic, vanillic, caffeic, protocatechuic, syringic, and p-coumaric acids, oleuropein and quercetin . Caffeic and protocatechuic acids (0.3 mg/ml) inhibited the growth of E . coli and K . pneumoniae . The same compounds apart from syringic acid (0.5 mg/ml) completely inhibited the growth of B . cereus . Oleuropein, and p-hydroxy benzoic, vanillic and p-coumaric acids (0.4 mg/ml) completely inhibited the growth of E . coli, K . pneumoniae and B . cereus . Vanillic and caffeic acids (0.2 mg/ml) completely inhibited the growth and aflatoxin production by both A . flavus and A . parasiticus, whereas the complete inhibition of the moulds was attained with 0.3 mg/ml p-hydroxy benzoic, protocatechuic, syringic, and p-coumaric acids and quercetin. Int J Immunopharmacol, 1997 Nov-Dec, 19(11-12), 629 - 44 The immunopharmacology of head and neck cancer: an update; Hadden JW; Patients with head and neck squamous cell cancer have cell-mediated immune defects and anergy, which progress with disease . T-lymphocytopenia and dysfunction, monocyte dysfunction, prostaglandins, antigen-antibody complexes, serum and cell suppressive factors, radiation therapy and poor nutrition with zinc deficiency all contribute . Nevertheless, cell-mediated immunoreactivity to tumor is manifest in the majority of the patient's blood and regional nodes, and in the tumor itself by tumor-infiltrating lymphocytes . Lymphocytes from these sources cloned in the presence of interleukin-2 +/- tumor extracts show relatively specific cytotoxicity against squamous cell cancer . Humoral immunity is intact, and increased IgA and IgE levels and antibodies reactive to tumor antigens are common . Tumor-associated antigens detected in serum and tumor include carcinoembryonic antigen, tumor polypeptide antigen, squamous cell cancer antigens, tumor antigen-4 and various mucin antigens . The mucin antigens, in particular, can elicit T-cell responses . Humoral reactivity to such antigens is manifest in circulating immune complexes and immunoglobulin coating of tumor surfaces . Immunotherapeutic efforts in head and neck squamous cell cancer should logically employ T-cell adjuvants, contrasuppression and immunorestoration . Non-specific stimulation with bacille Calmette-Guerin (BCG), levamisole and other agents has not been successful . Encouraging results have been observed in limited trials with indomethacin and plasmapheresis . Early trials with local administration of low dosages of interferon-alpha, natural interleukin-2 and a natural interleukin mixture have produced partial and complete regressions with no toxicity and with intense leukocyte infiltration indicating cellular immunity . Efforts are needed to define the mechanisms and the antigens involved in these reactions . On the contrary, treatments with high dosages of recombinant interferon-alpha and interleukin-2 have yielded few responses and considerable toxicity . Combination strategies are discussed which may improve upon these initial immunotherapeutic effects of these low dose trials. J Biol Chem, 1998 Jul 24, 273(30), 19097 - 101 Genetically engineered zinc-chelating adenylate kinase from Escherichia coli with enhanced thermal stability; Perrier V et al.; In contrast with adenylate kinase from Gram-negative bacteria, the enzyme from Gram-positive organisms harbors a structural Zn2+ bound to 3 or 4 Cys residues in the structural motif Cys-X2-Cys-X16-Cys-X2-Cys/Asp . Site-directed mutagenesis of His126, Ser129, Asp146, and Thr149 (corresponding to Cys130, Cys133, Cys150, and Cys153 in adenylate kinase from Bacillus stearothermophilus) in Escherichia coli adenylate kinase was undertaken for determining whether the presence of Cys residues is the only prerequisite to bind zinc or (possible) other cations . A number of variants of adenylate kinase from E . coli, containing 1-4 Cys residues were obtained, purified, and analyzed for metal content, structural integrity, activity, and thermodynamic stability . All mutants bearing 3 or 4 cysteine residues acquired zinc binding properties . Moreover, the quadruple mutant exhibited a remarkably high thermal stability as compared with the wild-type form with preservation of the kinetic parameters of the parent enzyme. Tuber Lung Dis, 1997, 78(1), 57 - 66 Progression of chronic pulmonary tuberculosis in mice aerogenically infected with virulent Mycobacterium tuberculosis; Rhoades ER et al.; There are several critical differences in the pulmonary granulomatous response to Mycobacterium tuberculosis between the mouse and other animal models such as the guinea pig or rabbit . One key difference is a conspicuous lack of central caseating necrosis in pulmonary lesions of immunologically intact mice . To determine whether normal mice could develop such pathology in response to highly virulent clinical isolates of M . tuberculosis, C57BL/6 mice were infected aerogenically with varying doses of three different strains, and the development of a granulomatous response was followed for as long as a year . Whereas such conditions failed to induce caseating necrosis in the lungs of these mice, all of the infections induced a granulomatous response which progressed similarly . We present here a descriptive report of the gross pathological progression of tuberculosis in the lungs of the mice . In each case, the disease progressed in five discrete stages, which were delineated on the basis of several criteria including the extent of granulomatous involvement, the cell types present, the degree of lymphocyte organization, and the presence of destructive sequelae such as airway epithelium erosion and airway debris . Quicker progression of disease along these five stages was induced by increasing the size of the inoculum or by the more virulent mycobacterial strains . The infections with the virulent strains were not resolved, and the later stages of the granulomatous response coincided with an increasing bacillary load and a loss of organized lymphocytes in the infected lungs which ultimately resulted in the death of the host . These results indicate that although C57BL/6 mice do not manifest a caseating form of pulmonary tuberculosis, they manifest an equally pathogenic granulomatous response which appears as a chronic interstitial fibrosing response that fails to contain the infection at a time that organized lymphocyte involvement wanes in the lung. Tuber Lung Dis, 1997, 78(1), 47 - 55 Effect of cytokine modulation by thalidomide on the granulomatous response in murine tuberculosis; Moreira AL et al.; SETTING: Experimental murine tuberculosis . OBJECTIVE: To evaluate the effect of cytokine modulation by thalidomide on the progression of the lung granulomatous response following aerosol tuberculosis infection in mice . DESIGN: Mice infected by the respiratory route with 200-500 viable Mycobacterium tuberculosis Erdman were treated with daily subcutaneous injections of thalidomide (30 mg/kg) or saline for 4 weeks . The bacillary load, granulomatous response and cytokine production in the lungs were evaluated . RESULTS: Aerosol M . tuberculosis infection resulted in a progressive granulomatous response in the lungs . At 28 days after infection, large granulomata with central necrosis and no apoptosis were observed . The infection induced high serum and lung cytokine mRNA levels . Thalidomide treatment resulted in a significant reduction in tumor necrosis factor-alpha, interleukin 6 (IL-6) and IL-10 protein levels (blood) and mRNA expression (lungs) . IL-12 and interferon-gamma were unaffected . The lungs of thalidomide-treated mice had smaller granulomata with apoptotic cells and no necrosis . Thalidomide treatment did not change the bacillary load . CONCLUSION: Thalidomide immunomodulation reduces inflammatory cytokines and concomitant lung pathology following acute aerosol M . tuberculosis infection, without increasing the bacillary load. Tuber Lung Dis, 1997, 78(1), 13 - 9 A novel repeat sequence specific to Mycobacterium tuberculosis complex and its implications; Lee TY et al.; Restriction fragment length polymorphism analysis of Korean clinical isolates of Mycobacterium tuberculosis using a 245 bp fragment of IS6110 revealed a conserved 3.5 kb Pvull fragment . Attempts to clone this 3.5 kb fragment resulted in the serendipitous discovery of a novel repeat sequence present within a separate 3.5 kb Pvull genomic fragment . Nucleotide sequencing of a 823 bp region containing the putative repeat sequence revealed the presence of three small direct repeats, three palindromes and a 453 bp region that was analogous to 455 bp of a M . tuberculosis sequence previously reported . The presence of this 453 bp repeat sequence was demonstrated in standard mycobacterial strains belonging to the M . tuberculosis complex, including the H37Rv, H37Ra, Erdman, and Canetti strains and M . bovis and M . bovis bacille Calmette-Guerin (BCG) . Other mycobacterial species (M . kansasii, M . smegmatis, M . simiae, M . fortuitum, M . scrofulaceum, M . intracellulare, M . avium, and M . haemophilum) did not contain this sequence, suggesting that the 453 bp repeat sequence was specific to the M . tuberculosis complex . Of the 13 Korean and 12 other clinical isolates of M . tuberculosis tested, all contained three to four copies of the repeat sequence . The Southern blot patterns of the various M . tuberculosis strains allowed classification into five different groups . The most frequent pattern was the 'BCG-type' (4.7, 3.5, and 2.4 kb bands); the second most frequent pattern was the '4-band-type' (13, 4.7, 3.5, and 2.4 kb), observed only in the Korean clinical isolates, and the third most common pattern was the M . tuberculosis H37Rv/H37Ra/M . bovis-type (13, 4.7, and 3.5 kb bands) . Upstream sequences indicate proximity to the rhamnose biosynthesis (rfb) cluster of M . tuberculosis . Our results indicate that the repeat sequence may be useful for the design of probe and polymerase chain reaction primers for the identification and epidemiological testing of members of the M . tuberculosis complex. Biochemistry, 1998 Jul 14, 37(28), 10173 - 80 Spectroscopic characterization of a binuclear metal site in Bacillus cereus beta-lactamase II; Orellano EG et al.; The zinc metalloenzyme beta-lactamase II (betaLII) from Bacillus cereus has been overexpressed in Escherichia coli as a fusion protein with glutathione-S-transferase, and the metal binding properties of recombinant betaLII toward Zn(II) and Co(II) have been studied by fluorescence and activity measurements . The apoenzyme is able to bind two metal ion equivalents, which confer on betaLII its maximum enzymatic efficiency . The enzyme is partially active with one metal ion equivalent . The diCo(II) and a mixed Zn(II)Co(II) derivative of betaLII were obtained and probed by electronic and paramagnetic NMR spectroscopy . In the high-affinity site, the metal is bound to three His residues and a solvent molecule, adopting a tetrahedral geometry . A Cys, a His, and an Asp residue are coordinated to the low-affinity metal site, together with two or three solvent molecules . This coordination polyhedron resembles the binuclear metal site of the Bacteroides fragilis beta-lactamase {Concha, N., Rasmussen, B . A., Bush, K., and Herzberg, O . (1996) Structure 4, 823-836; Carfi, A., Duee, E., Paul-Soto, R., Galleni, M., Frere, J . M., and Dideberg, O . (1998) Acta Crystallogr . D54, 47-57} but differs from that resulting from the X-ray study of betaLII {Carfi, A., Pares, S., Duee, E., Galleni, M., Duez, C., Frere, J . M., and Dideberg, O . (1995) EMBO J . 14, 4914-4921} . These results suggest that this binuclear metal site may be a general feature of metallo-beta-lactamases. Nat Struct Biol, 1998 Jul, 5(7), 585 - 92 The X-ray structure of a cobalamin biosynthetic enzyme, cobalt-precorrin-4 methyltransferase; Schubert HL et al.; Biosynthesis of the corrin ring of vitamin B12 requires the action of six S-adenosyl-L-methionine (AdoMet) dependent transmethylases, closely related in sequence . The first X-ray structure of one of these, cobalt-precorrin-4 transmethylase, CbiF, from Bacillus megaterium has been determined to a resolution of 2.4 A . CbiF contains two alphabeta domains forming a trough in which S-adenosyl-L-homocysteine (AdoHcy) binds . The location of AdoHcy and a number of conserved residues, helps define the precorrin binding site . A second crystal form determined at 3.1 A resolution highlights the flexibility of two loops around this site . CbiF employs a unique mode of AdoHcy binding and represents a new class of transmethylase. Curr Microbiol, 1998 Aug, 37(2), 80 - 7 Genetic diversity of Bacillus cereus/B . thuringiensis isolates from natural sources; Helgason E et al.; The genetic diversity and relationships among 154 Bacillus cereus/B . thuringiensis isolates recovered from soil samples from five geographic areas in Norway were investigated with multilocus enzyme electrophoresis (MEE) . Cluster analysis revealed two major groups (designated cluster I and cluster II) separated at genetic distance greater than 0.55 . Cluster I included 62 electrophoretic types (ETs) originating from all five locations, whereas, in cluster II, all but one isolate were from the same location . The isolates were also serotyped with B . thuringiensis flagellar antisera, and 28 distinct serotypes were identified . In general, serotyping did not show correlation to the genetic diversity of the isolates . The presence of IS231- and IS240-like transposable elements was detected in 14% of the strains of cluster II only . Parasporal crystals were observed in three strains; ten other strains were toxic to Trichoplusia ni . We conclude that B . cereus/B . thuringiensis from soil exhibit a high degree of recombination. Int J Tuberc Lung Dis, 1998 Jul, 2(7), 575 - 9 Hepatosplenic tuberculosis: a cause of persistent fever during recovery from prolonged neutropenia; Chakrabarti S et al.; SETTING: Hepatosplenic abscesses in neutropenic patients, especially during the recovery phase, are almost always attributed to fungal infections . We report similar lesions due to Mycobacterium tuberculosis in neutropenic patients in a tertiary care centre in India . OBJECTIVE: To characterize the features of hepatosplenic tuberculosis in neutropenic patients . DESIGN: Retrospective comparison of disease pattern and response to treatment of hepatosplenic tuberculosis in febrile neutropenia patients (four of 30 with severe prolonged neutropenia) and in non neutropenic patients diagnosed during the same 12-month period (n = 4, control group) . RESULTS: The disease in the neutropenic patients typically presented during the recovery phase of neutropenia, with ultrasonic abnormalities similar to those seen in hepatosplenic fungal infections . In contrast to the marked organomegaly and typical granulomatous response found in the control group, the disease in the neutropenic patients was characterised by an absence of organomegaly, non-involvement of other sites, poor inflammatory response and a high bacillary load . The initial response to therapy was satisfactory in both groups . CONCLUSION: Tuberculosis needs to be considered in the diagnostic work-up of hepatosplenic abscesses that occur during the recovery phase of neutropenia. J Biol Chem, 1998 Jul 17, 273(29), 18052 - 9 Dimeric tyrosyl-tRNA synthetase from Bacillus stearothermophilus unfolds through a monomeric intermediate . A quantitative analysis under equilibrium conditions; Park YC et al.; Tyrosyl-tRNA synthetase from Bacillus stearothermophilus comprises an N-terminal domain (residues 1-319), which is dimeric and forms tyrosyladenylate, and a C-terminal domain (residues 320-419), which binds the anticodon arm of tRNATyr . The N-terminal domain has the characteristic fold of the class I aminoacyl-tRNA synthetases . The unfolding of the N-terminal domain by urea at 25 degreesC under equilibrium conditions was monitored by its intensities of light emission at 330 and 350 nm, the ratio of these intensities, its ellipticity at 229 nm, and its partition coefficient, in spectrofluorometry, circular dichroism, and size-exclusion chromatography experiments, respectively . These experiments showed the existence of an equilibrium between the native dimeric state of the N-terminal domain, a monomeric intermediate state, and the unfolded state . The intermediate was compact and had secondary structure, and its tryptophan residues were partially buried . These properties of the intermediate and its inability to bind 1-anilino-8-naphthalenesulfonate showed that it was not in a molten globular state . The variation of free energy deltaG(H2O) and its coefficient m of dependence on the concentration of urea were, respectively, 13.8 +/- 0.2 kcal.mol-1 and 0.9 +/- 0.1 kcal.mol-1.M-1 for the dissociation of the native dimer and 13.9 +/- 0.6 kcal.mol-1 and 2.5 +/- 0.1 kcal.mol-1.M-1 for the unfolding of the monomeric intermediate. J R Soc Med, 1998 Mar, 91(3), 133 - 4 Exacerbation of atopic dermatitis after bacillus Calmette-Guérin vaccination; Dalton SJ et al.; In two children with atopic dermatitis, routine vaccination with bacillus Calmette-Guerin (BCG) was followed by severe exacerbation of skin disease . If the sequence is cause and effect, a possible mechanism is stimulation of a Th2 lymphocyte cytokine profile by the vaccine, with migration of activated lymphocytes to inflamed skin . In children with active atopic dermatitis, BCG vaccination is best deferred until remission. Plasmid, 1998 Jul, 40(1), 30 - 43 Kinetics of conjugative transfer: a study of the plasmid pXO16 from Bacillus thuringiensis subsp . israelensis; Andrup L et al.; The aggregation-mediated conjugation system of Bacillus thuringiensis subsp . israelensis, encoded by the 200-kb plasmid pXO16, is highly potent in transferring itself and efficient in mobilizing other nonconjugative plasmids . The present study reveals some salient features of this conjugation system . Our observations can be summarized as follows: (i) The conjugative transfer takes about 3(1/2) to 4 min . For a 200-kb plasmid this corresponds to about 1 kb per second . (ii) The ability to transfer the plasmid seems to be evenly distributed among the donors . (iii) Functionally, the mating complex was found to consist of one donor and one recipient cell, even though aggregates comprising thousands of interconnected cells are formed . (iv) Having donated the plasmid, the donor needs a "period of recovery" of about 10 min before it can redonate the plasmid . (v) Secondary transfer, i.e., transfer from newly formed transconjugants, is delayed about 40 min . This maturation time exceeds the generation time, and it may indicate that to display donor activity, a surface protein (the aggregation substance) has to be uniformly incorporated into the cell wall . Lastly, we found that when the experiments were sufficiently short and when the recipient cells were in excess compared with the donors, the process of conjugation could be reasonably described by a kinetic model analogous to the Michaelis-Menten model for enzyme catalysis . This allowed us to estimate (vi) the maximal conjugation rate to be about 0.05 transconjugant per donor per minute, and (vii) the Km value, i.e., the concentration of recipient that results in half of the maximal conjugation rate, to be about 4 x 10(6) recipients/ml . Arch Esp Urol, 1998 May, 51(4), 389 - 90 {Hematuria, dysuria, pollakiuria and general malaise, in a patient treated with BCG instillations}; Medina Perez M et al.; OBJECTIVE: To report a case of granulomatous cystitis in a patient receiving bacillus Calmette-Guerin intravesical therapy for urothelial carcinoma in situ . METHODS: A 58-year-old man undergoing BCG intravesical therapy for urothelial carcinoma in situ presented symptoms of intense cystitis . Cystoscopy was performed and several bladder cold biopsies were obtained . RESULTS: Histopathological analysis demonstrated epithelioid granulomas . CONCLUSION: Cystitis arising from BCG therapy is defined as drug-induced or BCG-induced cystitis . Intense cystitis and malaise are a serious complication since it is not possible to distinguish patients with a simple uncomplicated local reaction from those who will develop progressive systemic infection . Cystoscopy and biopsy can be helpful in determining the nature of the condition and are recommended. Eur J Biochem, 1998 Apr 15, 253(2), 480 - 4 Pyrrole-2-carboxylate decarboxylase from Bacillus megaterium PYR2910, an organic-acid-requiring enzyme; Omura H et al.; Inducible pyrrole-2-carboxylate decarboxylase, which catalyzes the decarboxylation of pyrrole-2-carboxylate to pyrrole and CO2 in stoichiometric amounts, was purified from Bacillus megaterium PYR2910 . The purity of the enzyme was shown by SDS/PAGE and gel-permeation HLPC . The enzyme has a molecular mass of approximately 98 kDa and consists of two identical subunits . It is highly specific for pyrrole-2-carboxylate, and also catalyzes the reverse reaction, the carboxylation of pyrrole . A unique feature of this enzyme is its requirement of an organic acid, such as acetate, propionate, butyrate or pimelate . A possible catalytic mechanism including a cofactor function of organic acid is discussed. J Infect Dis, 1998 Jul, 178(1), 138 - 46 Cellular immune responses to four doses of percutaneous bacille Calmette-Guérin in healthy adults; Lowry PW et al.; To explore the hypothesis that low-dose immunization might induce preferential Th1 cell immunity, 76 adults were vaccinated with one of four doses of bacille Calmette-Guerin (BCG): The doses contained very low (1.6 x 10(5) cfu), low (3.2 x 10(6) cfu), standard (1.6 x 10(8) cfu), or high (3.2 x 10(8) cfu) levels of BCG . Delayed-type hypersensitivity responses occurred 8 weeks after vaccination in 10% of persons given very low or low doses of BCG, compared with 95% and 100% of persons given standard or high doses, respectively . Lymphoproliferative responses, which were increased only for high-dose vaccinees, peaked 2 weeks after vaccination and were directed chiefly against Mycobacterium tuberculosis-secreted proteins, particularly the antigen 85 complex . Significant increases in mycobacteria-specific interferon-gamma expression were present 16 weeks after vaccination only for persons given standard or high doses of BCG . Percutaneous BCG appears capable of inducing a temporary Th1-like immune response, but standard or higher dosages are required. J Clin Microbiol, 1998 Jul, 36(7), 2138 - 9 Bacillus thuringiensis subsp . konkukian (serotype H34) superinfection: case report and experimental evidence of pathogenicity in immunosuppressed mice; Hernandez E et al.; We present a case of severe war wounds infected by Bacillus thuringiensis serotype H34 and describe the experimental protocol used to demonstrate its ability to infect mice after cutaneous inoculation . This case is interesting because B . thuringiensis is considered to be a contaminant in laboratories and receives inadequate attention. Nucleic Acids Res, 1998 Jul 15, 26(14), 3348 - 9 BfiI, a restriction endonuclease from Bacillus firmus S8120, which recognizes the novel non-palindromic sequence 5'-ACTGGG(N)5/4-3'; Vitkute J et al.; A new type IIS restriction endonuclease Bfi I hasbeen partially purified from Bacillus firmus S8120 . Bfi I recognizes the non-palindromic hexanucleotide sequence 5'-ACTGGG(N)5/4-3' and makes a staggered cut at the fifth base pair downstream of the recognition sequence on the upper strand, producing a single base 3' protruding end. Carbohydr Res, 1997 Dec, 305(3-4), 561 - 8 Transfer reactions catalyzed by cyclodextrin glucosyltransferase using 4-thiomaltosyl and C-maltosyl fluorides as artificial donors; Bornaghi L et al.; Cyclodextrin glycosyltransferase enzyme from Bacillus circulans catalyzed the effective conversion of 4-thio-alpha-maltosyl fluoride into cyclo-alpha-(1-->4(2))-thiomalto -tetraoside, -pentaoside, -hexaoside and linear hemithiomaltooligosaccharides . However, under the same conditions, C-maltosyl fluoride afforded only linear modified maltotetraose, maltohexaose and maltooctaose in moderate yield. Carbohydr Res, 1997 Dec, 305(3-4), 393 - 400 Enzymatic synthesis, isolation, and analysis of novel alpha- and beta-galactosyl-cycloisomalto-octaoses; Koizumi K et al.; Novel branched cycloisomalto-octaoses (CI8s) were enzymatically synthesized by transgalactosylation with alpha-galactosidase from coffee bean and beta-galactosidase preparations from Penicillium multicolor and Bacillus circulans, using melibiose and lactose as donor substrates, and CI8 which is a cyclic homogeneous oligosaccharide composed of eight glucose units bound by alpha-(1-->6)-linkages, as an acceptor . alpha-Galactosyl-CI8s and beta-galactosyl-CI8s obtained were isolated and purified by HPLC . Their structures were elucidated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDITOFMS) and NMR spectroscopy. Biosci Biotechnol Biochem, 1998 May, 62(5), 1028 - 30 Thermostable neutral protease resembling thermolysin derived from Bacillus brevis MIB001; Takii Y et al.; A microbe producing a protease with strong thermostability that was released extracellularly was isolated from soil . The isolate, MIB001, grew at from 15 to 51 degrees C and pH 5.1-8.8 and was tentatively identified as a strain of Bacillus brevis . Rabbit antisera raised against a pure preparation of the protease did not cross-react with thermolysin or neutral metalloprotease from Bacillus stearothermophilus KP1236. Appl Environ Microbiol, 1998 Jul, 64(7), 2723 - 5 Insecticidal activity of Bacillus laterosporus; Orlova MV et al.; The Bacillus laterosporus strains 921 and 615 were shown to have toxicity for larvae of the mosquitoes Aedes aegypti, Anopheles stephensi, and Culex pipiens . The larvicidal activity of B . laterosporus was associated with spores and crystalline inclusions . Purified B . laterosporus 615 crystals were highly toxic for Aedes aegypti and Anopheles stephensi. Appl Environ Microbiol, 1998 Jul, 64(7), 2497 - 502 Development of a streptavidin-conjugated single-chain antibody that binds Bacillus cereus spores; Koo K et al.; Control of microorganisms such as Bacillus cereus spores is critical to ensure the safety and a long shelf life of foods . A bifunctional single chain antibody has been developed for detection and binding of B . cereus T spores . The genes that encode B . cereus T spore single-chain antibody and streptavidin were connected for use in immunoassays and immobilization of the recombinant antibodies . A truncated streptavidin, which is smaller than but has biotin binding ability similar to that of streptavidin, was used as the affinity domain because of its high and specific affinity with biotin . The fusion protein gene was expressed in Escherichia coli BL21 (DE3) with the T7 RNA polymerase-T7 promoter expression system . Immunoblotting revealed an antigen specificity similar to that of its parent native monoclonal antibody . The single-chain antibody-streptavidin fusion protein can be used in an immunoassay of B . cereus spores by applying a biotinylated enzyme detection system . The recombinant antibodies were immobilized on biotinylated magnetic beads by taking advantage of the strong biotin-streptavidin affinity . Various liquids were artificially contaminated with 5 x 10(4) B . cereus spores per ml . Greater than 90% of the B . cereus spores in phosphate buffer or 37% of the spores in whole milk were tightly bound and removed from the liquid phase by the immunomagnetic beads. Appl Environ Microbiol, 1998 Jul, 64(7), 2490 - 6 Construction and expression of a bifunctional single-chain antibody against Bacillus cereus p6ores; Koo K et al.; The variable-region genes of monoclonal antibody against Bacillus cereus spores were cloned from mouse hybridoma cells by reverse transcription-PCR . The heavy- and light-chain variable-region genes were connected by a 45-base linker DNA to allow folding of the fusion protein into a functional tertiary structure . For detection of protein expression, a 10-amino-acid strep tag (biotin-like peptide) was attached to the C terminus of recombinant antibody as the reporter peptide . The single-chain antibody construct was inserted into the expression vector and expressed in Escherichia coli under the control of the T7 RNA polymerase-T7 promoter expression system . The expressed single-chain antibody was detected on Western blots by using a streptavidin-conjugated enzyme system . This small recombinant antibody fragment (ca . 28,000 Da by calculation) had B . cereus spore binding ability and antigen specificity similar to those of its parent native monoclonal antibody. Appl Environ Microbiol, 1998 Jul, 64(7), 2479 - 84 A two-component monooxygenase catalyzes both the hydroxylation of p-nitrophenol and the oxidative release of nitrite from 4-nitrocatechol in Bacillus sphaericus JS905; Kadiyala V et al.; Bacteria that metabolize p-nitrophenol (PNP) oxidize the substrate to 3-ketoadipic acid via either hydroquinone or 1,2,4-trihydroxybenzene (THB); however, initial steps in the pathway for PNP biodegradation via THB are unclear . The product of initial hydroxylation of PNP could be either 4-nitrocatechol or 4-nitroresorcinol . Here we describe the complete pathway for aerobic PNP degradation by Bacillus sphaericus JS905 that was isolated by selective enrichment from an agricultural soil in India . Washed cells of PNP-grown JS905 released nitrite in stoichiometric amounts from PNP and 4-nitrocatechol . Experiments with extracts obtained from PNP-grown cells revealed that the initial reaction is a hydroxylation of PNP to yield 4-nitrocatechol . 4-Nitrocatechol is subsequently oxidized to THB with the concomitant removal of the nitro group as nitrite . The enzyme that catalyzed the two sequential monooxygenations of PNP was partially purified and separated into two components by anion-exchange chromatography and size exclusion chromatography . Both components were required for NADH-dependent oxidative release of nitrite from PNP or 4-nitrocatechol . One of the components was identified as a reductase based on its ability to catalyze the NAD(P)H-dependent reduction of 2,6-dichlorophenolindophenol and nitroblue tetrazolium . Nitrite release from either PNP or 4-nitrocatechol was inhibited by the flavoprotein inhibitor methimazole . Our results indicate that the two monooxygenations of PNP to THB are catalyzed by a single two-component enzyme system comprising a flavoprotein reductase and an oxygenase. Biochem Biophys Res Commun, 1998 Jun 29, 247(3), 659 - 62 Peroxide reductase activity of NADH dehydrogenase of an alkaliphilic Bacillus in the presence of a 22-kDa protein component from Amphibacillus xylanus; Koyama N et al.; The NADH oxidase of Amphibacillus xylanus shows high NADH-peroxide reductase activity for hydrogen peroxide and alkyl hydroperoxides in the presence of a 22-kDa disulfide-containing protein component (Y . Niimura, L . B . Poole, and V . Massey, J . Biol.Chem . 270, 25645-25650, 1995) . It was found that the membrane-bound NADH dehydrogenase of an alkaliphilic Bacillus (YN-1) involved in the respiratory chain also exhibits reductase activity for hydrogen peroxide and cumene hydroperoxide in the presence of the 22-kDa component from Amphibacillus xylanus . Vmax values for these substrates were as high as those of the NADH oxidase of A . xylanus . Although the 38-kDa protein produced by trypsin treatment of NADH dehydrogenase retains NADH dehydrogenase activity, it exhibited no peroxide reductase activity in the presence of the 22-kDa component from A . xylanus . The NADH dehydrogenase of YN-1 might not only catalyze electron flow from NADH to the respiratory chain, but also function for scavenging peroxide. J Invertebr Pathol, 1998 Jul, 72(1), 73 - 81 Processing of delta-endotoxin of Bacillus thuringiensis subsp . kurstaki HD-1 in Heliothis armigera midgut juice and the effects of protease inhibitors; Shao Z et al.; Bombyx mori was found to be more sensitive to the protoxins of HD-1 than Heliothis armigera . SDS-PAGE analysis showed that a large amount of activated toxin was yielded from protoxin by B . mori gut juice while little was yielded by H . armigera . Further degradation of activated toxin was observed in H . armigera midgut juice detected by SDS-PAGE . pH influenced the proteolytic activity of the midgut juice significantly, but there was no obvious effect of pH on the degradation of activated toxin . Specific inhibitor study revealed the presence of trypsin, chymotrypsin, and elastase in the midgut juice . TLCK, TPCK, elastatinal and some general serine protease inhibitors successfully prevented the excessive degradation of protoxin in H . armigera midgut juice . Chymotrypsin inhibitors showed strong inhibitory effects against the further degradation of activated toxin, indicating that chymotrypsin played a major role in the process . It was presumed that the excessive degradation of protoxin in H . armigera midgut juice was responsible for the low sensitivity of the insect to Bt . Further study demonstrated that the excessive degradation in vitro was triggered by SDS treatment . However, all of the tested serine protease inhibitors expressed synergism with protoxin against H . armigera larvae, suggesting that the excessive degradation of protoxin may occur in vivo to some extent and may be triggered by receptor binding of activated toxin . J Invertebr Pathol, 1998 Jul, 72(1), 9 - 20 Histopathological effects of Bacillus thuringiensis on the alimentary canal of the sheep louse, Bovicola ovis; Hill CA et al.; Sequential observations were made of the ultrastructural effects of Bacillus thuringiensis (Bt) subsp . kurstaki strain WB3S16 on midgut epithelial cells of the sheep biting louse, Bovicola ovis, after the lice were fed, ad libitum, a powdered preparation of Bt spores, delta-endotoxin crystals, and lysed cellular components . Light microscope observations revealed cytopathological changes to the midgut epithelial cells 4 h postfeeding . Transmission electron micrographs showed that the microvilli of the midgut epithelial cells became disrupted 4-8 h postfeeding, after which the cells became vacuolated and swollen, and the cell organelles lost definition and disappeared . Paralysis and death of B . ovis occurred between 8 and 12 h postfeeding, coincident with midgut cells lysis and release of cellular contents into the midgut lumen . The histopathological effects reported here are similar to those reported in lepidopteran and coleopteran larvae affected by the delta-endotoxin crystal proteins . The constituent fractions of the Bt preparation were tested for toxicity to B . ovis using a feeding bioassay . Native delta-endotoxin crystals were not toxic to B . ovis and remained intact in the midgut of the insect . There was no evidence that the native Bt crystal was involved in the cytopathology and death of the lice . However, in vitro solubilized delta-endotoxin crystal proteins were significantly toxic to B . ovis . In addition, a louse active toxin was associated with the Bt membranes and culture supernatant . Eur J Immunol, 1998 Jun, 28(6), 1762 - 72 Failure of P strain mice to respond to vaccination against schistosomiasis correlates with impaired production of IL-12 and up-regulation of Th2 cytokines that inhibit macrophage activation; Oswald IP et al.; In contrast to most inbred strains, P mice fail to develop significant resistance to Schistosoma mansoni infection as a result of vaccination with either radiation-attenuated cercariae or schistosome antigens plus Bacillus Calmette Guerin, and this failure correlates with defects in macrophage larvicidal activity . Supernatant fluids from antigen-treated in vitro cultures of splenocytes from vaccinated P mice demonstrate less macrophage stimulatory activity than do supernatants from cells of vaccine-responsive strains such as C57BL/6 . This is not due either to diminished production of the macrophage-activating cytokine IFN-gamma by P mice, or to a lesser responsiveness of macrophages from P mice to activation by IFN-gamma . Rather, P splenocytes produce two-to threefold higher amounts of IL-4 and IL-10, cytokines which down-regulate the cytotoxic potential of IFN-gamma-treated macrophages . Thus, the macrophage-activating potential of cytokine preparations from vaccinated P mice can be completely recovered by in vitro treatment with antibodies to IL-4 or IL-10 . Moreover, lower levels of IL-12, a cytokine involved in promoting development of Th1 responses, are produced by splenocytes from P mice as compared to C57BL/6 counterparts . These studies indicate that a genetic predisposition toward an impaired production of IL-12 and an increased production of down-regulatory Th2 cytokines correlate with low response to vaccination against S . mansoni. FEBS Lett, 1998 May 22, 428(1-2), 57 - 8 Contribution of arginine-82 and arginine-86 to catalysis of RNases from Bacillus intermedius (binase); Yakovlev GI et al.; To elucidate the functional role of Arg82 and Arg86 in the enzyme activity of binase, the extracellular ribonuclease of Bacillus intermedius, we used site-directed mutagenesis . On cleavage of various substrates the catalytic activity of binase mutant Arg86 Ala is 2.7 x 10(3) - 7.7 x 10(3) times less than that of the native enzyme . The decrease in activity is determined preferentially by the decrease in the molecular rate constant kcat with a relatively small change of enzyme-substrate affinity, characterized by Km . This is the expected result if Arg86 acts to lower the energy of a transition state of the reaction . The replacement of Arg82 by Ala causes a 5-19-fold activity decrease, depending on the substrate . We propose that this residue does not have a direct catalytic function in the molecular mechanism of the binase action and that the activity decrease of binase on the replacement of Arg82 by alanine is mediated by the effect of Arg82 on the pK of catalytic residues. J Biochem (Tokyo), 1998 Jul, 124(1), 45 - 50 Lysyl-tRNA synthetase from Bacillus stearothermophilus . Stopped-flow kinetic analysis of enzyme.lysyladenylate formation; Takita T et al.; Amino acid activation reaction of the lysyl-tRNA synthetase {L-lysine:tRNALys ligase (AMP forming); EC 6.1.1.6} from Bacillus stearothermophilus was studied fluorometrically by the stopped-flow method . The addition of L-lysine to the enzyme solution caused quenching of the protein fluorescence and the subsequent addition of ATP restored the quenched fluorescence {Takita et al . (1996) J . Biochem . 119, 680-689; Takita et al . (1997) 121, 244-250} . In the stopped-flow analysis, however, the former fluorescence change (quenching) could not be detected, while the latter change (restoration) was detectable . The L-lysine binding process was suggested to be much faster than the ATP binding process, being completed within the dead-time of the apparatus, ca . 3 ms . The hyperbolic dependence of kapp on the initial ATP concentration suggested that the ATP binding to the enzyme.L-lysine complex followed a two-step mechanism . Two L-lysine analogues that exhibit the qualitatively similar behavior to L-lysine in the fluorometric titration, L-lysine hydroxamate and L-lysine amide, were examined similarly . The two-step process was also suggested for these analogues, and the forward rate constant in the rate-determining step for L-lysine amide (221+/-7 s-1) was significantly larger than those for L-lysine (45.7+/-4.6 s-1) and L-lysine hydroxamate (14 . 5+/-1.7 s-1) at pH 8.0, 30 degrees C. Commun Dis Public Health, 1998 Jun, 1(2), 84 - 8 Complications of bacille Calmette-Guérin (BCG) vaccination and immunotherapy and their management; Grange JM; Complications of bacille Calmette-Guerin (BCG) vaccination are uncommon . Fewer than one in 1000 people vaccinated develop significant local reactions, and serious disseminated disease develops in fewer than one in a million . Localised complications--which include hypersensitivity reactions, abscesses at the injection site, and localised lymphadenopathy--are usually self limiting . They usually result from faulty technique, including the accidental intracutaneous injection of the stronger percutaneous vaccine, or poor selection of subjects for vaccination . Abscesses at the injection site usually respond to drainage and chemotherapy with isoniazid or erythromycin . Lymphadenopathy responds poorly to antimicrobial treatment and surgery may be needed for suppurating or discharging lesions to hasten recovery and give a good cosmetic result . Disseminated disease usually occurs in people with impaired immunity, in whom it is often fatal . BCG should never be given to people who are known to be infected with HIV, but the risk of complications in children born to HIV infected mothers is low . Disseminated disease can also result from intravesical instillation of BCG to treat bladder cancer, but this responds to antituberculosis chemotherapy. Eur Urol, 1998, 33(5), 457 - 63 Primary bladder carcinoma in situ: assessment of early BCG response as a prognostic factor; Orsola A et al.; OBJECTIVES: To evaluate the prognosis of primary bladder carcinoma in situ (CIS) according to the response to bacillus Calmette-Guerin (BCG) . Patients and METHODS: Twenty-six cases of primary CIS were treated with BCG . Mean, median and minimum follow-up periods were 47, 56 and 24 months . At 6 months, the patients were evaluated endoscopically and the response was classified as complete, partial or failure . RESULTS: Twenty-one patients (80.8%) showed complete response to BCG, 3 did so after a second course; 28.5% relapsed or progressed at a mean of 44 months . Five patients (19.2%) did not respond initially and all progressed in a period of 6 months . Early response to BCG was the only significant prognostic factor (p < 0.005) . CONCLUSIONS: A high- and a low-risk group of bladder CIS can be differentiated according to the response to BCG . CIS of the bladder has a poor prognosis, and the number of patients who developed progressive disease is significantly higher among the nonresponders. J Nat Prod, 1998 Jun 26, 61(6), 729 - 33 Enhancement of NO production in activated macrophages in vivo by an antimalarial crude drug, Dichroa febrifuga; Murata K et al.; The effect of an antimalarial crude drug, Dichroafebrifuga Lour . on nitric oxide (NO) production in bacillus Calmette Guerin-induced mouse peritoneal macrophages activated by lipopolysaccharide was investigated . The NO production was significantly enhanced by an oral administration of a MeOH extract of D . febrifuga . Febrifugine (1) was isolated as the main active compound, and the activation was dose-dependent in the dosage range of 0.1-1 mg/kg/day. Semin Respir Infect, 1998 Jun, 13(2), 100 - 8 Immunologic response and pathophysiology of Legionella infection; Friedman H et al.; Legionella pneumophila, the causative agent of legionnaires' disease, is a gram-negative pleomorphic bacillus and fastidious in its growth in artificial medium . These bacteria grow readily intracellularly, including growth in macrophages and other phagocytic cells . Humoral antibodies develop readily to these bacteria not only in infected patients, but also in persons who have had subclinical exposure . High-levels of serum antibodies may also occur in individuals who recover from infection . However, cell-mediated immunity based on lymphocytes reacting with the organisms and cytokines produced by such lymphocytes are important in resistance . Vaccines prepared from killed Legionella or their components readily induce cell-mediated immunity . Immune resistance to disease depends on lymphocyte-based immunity, activating cytokine formation, some of which activate macrophages to resist infection . Resistance to Legionella infection by experimental animals such as mice correlates with activation of macrophages, which can inhibit replication of the bacteria . Much recent experimental work has involved studies using inbred animals, including inbred mice genetically resistant to Legionella versus mice genetically susceptible . Detailed studies show that regulation of macrophage resistance versus susceptibility to infection is mediated by specific genetic mechanisms . Induction of cytokines by Legionella can activate immune cells, especially helper T cells . Th 1 type helper cells that produce type 1 class cytokines, such as interferon gamma and interleukin-2 (IL-2), are known to be important in cellular immunity to Legionella as well as to other opportunistic intracellular bacteria . In contrast, Th 2 type helper cells, which secrete type 2 class cytokines such as IL-4, IL-5, and IL-6, activate B lymphocytes to produce humoral antibodies important in resistance to extracellular bacteria which secrete toxins and extracellular factors as compared to intracellular bacteria such as Legionella . Although Legionella, similar to other ubiquitous opportunistic pathogens, can cause serious infection in immunocompromised individuals, these bacteria have many distinguishing characteristics, such as very rapid replication in macrophages from susceptible individuals . However, activated macrophages restrict the growth of these bacteria . Infection by Legionella, if recognized clinically, can be readily treated with appropriate antibiotics . Currently, many studies are in progress concerning the mechanism of pathogenicity and assessment of the molecular biologic mechanisms of protective immune responses to this bacterium, which causes serious infection in immunocompromised individuals. Science, 1998 Jun 26, 280(5372), 2129 - 32 Insecticidal toxins from the bacterium Photorhabdus luminescens; Bowen D et al.; Transgenic plants expressing Bacillus thuringiensis (Bt) toxins are currently being deployed for insect control . In response to concerns about Bt resistance, we investigated a toxin secreted by a different bacterium Photorhabdus luminescens, which lives in the gut of entomophagous nematodes . In insects infected by the nematode, the bacteria are released into the insect hemocoel; the insect dies and the nematodes and bacteria replicate in the cadaver . The toxin consists of a series of four native complexes encoded by toxin complex loci tca, tcb, tcc, and tcd . Both tca and tcd encode complexes with high oral toxicity to Manduca sexta and therefore they represent potential alternatives to Bt for transgenic deployment. Immunology, 1998 Mar, 93(3), 307 - 13 Inhibition of an established allergic response to ovalbumin in BALB/c mice by killed Mycobacterium vaccae; Wang CC et al.; Allergic disorders are mediated by T lymphocytes secreting T helper 2 (Th2) cytokines, interleukin-4 (IL-4) and interleukin-5 (IL-5), resulting in high levels of serum immunoglobulin E (IgE) and recruitment of eosinophils . One of the treatment strategies is to downregulate the Th2 component by inducing a T helper 1 (Th1) response to the relevant allergen, because Th1 and Th2 cytokines are thought to be mutually antagonistic . In this study, we examined the effects of Mycobacterium vaccae, a potent inducer of Th1 immunity, on allergic responses in a murine model . A single injection of M . vaccae into ovalbumin (OVA)-preimmunized BALB/c mice suppressed serum IgE over a wide dose range (10(7), 10(8) or 10(9) M . vaccae) . Further experiments, using 10(7) M . vaccae injected twice, showed that this treatment inhibited not only serum IgE, but also the potential for ovalbumin-induced IL-5 production by spleen cells . This non-specific ability of a mycobacterium to decrease Th2 activity, even when not presented together with the allergen, is in agreement with recent epidemiological studies on the impact of bacillus Calmette-Guerin (BCG) vaccination, and of other potent Th1 stimuli, on the incidence of atopy . The suppression of serum IgE and allergen-specific IL-5 synthesis by M . vaccae suggest that this organism is likely to have clinical application in the immunotherapy of allergy. Wei Sheng Wu Xue Bao, 1996 Aug, 36(4), 303 - 6 {Characterization of insecticidal crystal proteins of Bacillus thuringiensis subsp . chinensis CT-43}; Sun M et al.; Bacillus thuringiensis subsp . chinensis CT-43, no flagellum, produces various shaped parasporal crystals, which consist of 140000, 130000, and 65000 proteins . Based on two kinds of mutant, 140000 and 130000 crystal protein individually forms bipyramidal crystal and the 65000 protein forms cubidal crystal, and that the 140000 and 130000 protein is activated by trypsin into 55000 and 66000 proteins and 60000 protein, respectively . Bioassay were conducted to 3rd instar Plutella xylostella larvae with crystals, soluble crystal proteins, and activated crystal proteins, respectively, and it indicated that high toxic mutants can be obtained by curing low toxic crystal genes, and that the toxicity of crystals can be improved 16.3 to 58.4 times after solubilization. Wei Sheng Wu Xue Bao, 1996 Aug, 36(4), 295 - 302 {Distribution of Bacillus thuringiensis in soils of north and south of China}; Dai S et al.; 221 isolates of Bacillus thuringiensis were isolated in 1491 soil samples from North and South of China . H-serotypes and larvicidal characters of all Bt isolates have been identified . The rate of Bt-harbouring soil sample and the rate of Bt isolates in Northeast and Neimeng were in 12.6% and 17.2% respectively . Predominant serotypes were H4, H10, H3, H13, H5 and H29 . The most fertile Bt-harbouring area was the Heilongjiang Province with rate of Bt-harbouring sample of 21.4% and rate of Bt isolate of 29.4% . Rate of Bt-harbouring sample and rate of Bt isolate in Northwest area were 6.6% and 7.1% respectively . Main serotypes were H4, H5, H19, H10 and H3 . In four provinces of Southern China, both rates above were 18.6% and 29.5%, but frequency of Bt distribution was varied seriously in different distinct . Predominant serotypes in soils from Southern China were H3 and H5 . Results of bioassay showed that the percentage of strains high active to Heliothis armigera and Plogioidera versicolora were 1.6% and 1.1% in soils from North of China . In contrast to North of China, Bt strains active to H . armigera were 5.3% and none of Bt was effective to P . versicolora in South of