Int J Dermatol, 1991 Nov, 30(11), 780 - 4 Characterization of circulating lymphocytes by monoclonal antibodies in childhood and adult leprosy; Sehgal VN et al.; Peripheral blood lymphocyte assays using monoclonal antibodies were done in 66 patients with leprosy, consisting of 25 children and 41 adults . The results were statistically analyzed for correlations, if any, among the different age groups and matched controls . The results, however, failed to show any significant correlation, nor was it possible to draw any conclusion as to why the disease spectrum in children tends to be incomplete (ie, there is a low incidence of the highly bacilliferous form of disease expression). Pediatr Infect Dis J, 1991 Nov, 10(11), 832 - 6 Miliary tuberculosis in children: a review of 94 cases; Hussey G et al.; This is a retrospective review of the clinical, radiologic and laboratory features of 94 cases of childhood miliary tuberculosis seen during a 5-year period, 1985 to 1989 . A history of Bacillus Calmette-Guerin vaccination was documented in 88% of children . The median age at presentation was 10.5 months, 52% of cases occurring in those younger than 1 year . The presenting symptoms were nonspecific: cough (72%); fever (61%); loss of appetite and weight (40%); and diarrhea and vomiting (33%) . The main presenting signs were hepatomegaly (82%), splenomegaly (54%), lymphadenopathy (46%) and pyrexia (39%) . Most of the patients were malnourished and anergic . Meningitis occurred in 19% of patients and this was the only significant risk factor identified for mortality, the overall case fatality rate being 14% . The diagnosis in the vast majority was made on the clinical presentation supported by a classic miliary pattern on chest roentgenogram (91% of cases) . Mycobacterium tuberculosis was cultured in 33% of cases . In addition a review of hospital admissions from 1981 to 1989 revealed that annually miliary tuberculosis in children and adults accounted for 8.3 and 1.3%, respectively, of all tuberculosis admissions . This study confirms that miliary tuberculosis is a relatively common complication of tuberculosis in young children. J Neurol Neurosurg Psychiatry, 1991 Nov, 54(11), 989 - 92 Whipple's disease confined to the CNS presenting with multiple intracerebral mass lesions; Wroe SJ et al.; A patient with isolated cerebral Whipple's disease presented with signs of raised intracranial pressure and multiple ring enhancing intracerebral mass lesions evident on CT and MRI imaging . Characteristic intracellular bacilliform inclusions were identified in a brain biopsy . Clinical improvement followed treatment with parenteral antibiotics for two weeks and long term sulphamethoxazole-trimethoprim . As CNS relapse of Whipple's disease may occur after several years, long term treatment should include antibiotics that are able to cross the blood-brain barrier. Gan To Kagaku Ryoho, 1991 Nov, 18(14), 2369 - 74 {Superficial bladder cancer: prophylaxis of recurrence and progression}; Miyanaga N et al.; Superficial bladder cancer has a good prognosis compared with invasive bladder cancer . However, recurrence of the tumor is frequent and tumor stage and/or grade progress at the time of recurrence in many cases . Intravesical chemotherapy has been employed as a prophylactic method after trans-urethral resection (TUR) . Although intravesical chemotherapy has been proved to be effective in delaying the first recurrence of tumor after TUR, it cannot improve the ultimate prognosis of superficial bladder cancer . Many primary, solitary, non-invasive (Ta) and grade 1 tumors do not recur or progress in stage and grade . In these cases, prophylactic intravesical chemotherapy is not essential . Bacille Calmette-Guerin (BCG) should be considered superior overall, to any chemotherapeutic agents . Comparative studies will give information about the best clinical schedule for the treatment of superficial bladder cancer. Indian J Exp Biol, 1991 Nov, 29(11), 1031 - 4 Fed-batch fermentation studies with Bacillus thuringiensis H-14 synthesising endotoxin; Kuppusamy M et al.; Cell yield and toxicity of B . thuringiensis H-14 was improved markedly by adopting a simple fed-batch fermentation technique based on controlling glucose concentration . Maintenance of steady glucose concentration (0.3-0.5%) in the culture medium was achieved by the continuous addition of concentrated glucose solution . Addition of glucose at 3 g/hr/l of culture starting from 3rd hr till 16th hr of fermentation was found to yield cell densities of 80 g/l (wet weight) which represented a nearly 3-fold increase over the batch mode . A fivefold increase in toxicity was obtained by fed-batch fermentation . Cultivation of B . thuringiensis H-14 to high cell densities had no negative effect on sporulation and toxin synthesis . The rate of pH drop and dissolved oxygen level were within manageable limits. Mol Gen Mikrobiol Virusol, 1991 Nov, (11), 20 - 2 {Search for producers of restriction endonucleases with a given specificity}; Repin VE et al.; Six site-specific restriction endonucleases were isolated from Bacillus thuringiensis strains chosen of 58 strains ought purposefully for production of restriction enzymes . All six strains produce isoschisomers of Sau3A. Mol Microbiol, 1991 Nov, 5(11), 2799 - 806 The C-terminal domain of the toxic fragment of a Bacillus thuringiensis crystal protein determines receptor binding; Honee G et al.; The insecticidal crystal proteins of Bacillus thuringiensis show a high degree of specificity . In vitro binding studies with several crystal proteins demonstrated a correlation between toxicity and binding to receptors of larval midgut epithelial cells . In order to study the domain-function relationships of the toxic fragment, hybrid crystal proteins based on CryIA(b) and CryIC were constructed . Two out of 11 hybrid proteins constructed exhibited insecticidal activity . Both dispalyed an insecticidal spectrum similar to that of the parental crystal protein from which the C-terminal part of the toxic fragment originated . In addition, in vitro binding studies directly demonstrated the involvement of the C-terminal part of the toxic fragment in receptor binding . These results demonstrate that the C-terminal part of the toxic fragment determines specific receptor binding, which in turn determines, to a large extent, the insect specificity. Biotechnol Prog, 1991 Nov-Dec, 7(6), 516 - 25 Characterization of kappa-carrageenan gels used for immobilization of Bacillus firmus; Moon SH et al.; In this study, aimed at a biochemical and physical characterization of kappa-carrageenan gels used for entrapment of Bacillus firmus NRS 783 (a superior producer of an alkaline protease), effects of carrageenan concentration, gelation temperature, initial cell loading, and strength of the curing agent (KCl) on the properties of cell-free and cell-laden gels were examined . The physical properties of the differently prepared gels that were examined included density, free volume fraction, mechanical strength, and change in gel volume during gel curing . The biochemical characteristics studied included viability of gel-entrapped cells, cell leakage from cell-laden gels, and cell penetration into cell-free gels . For the range of carrageenan contents investigated {between 2% and 5% (w/v)}, the mechanical strength of the gels with/without KCl curing was observed to increase with an increase in carrageenan content of gels . The mechanical strength of each gel increased substantially upon extensive curing . Free volume fractions in excess of 0.8 were observed for all gels . Of cells that were viable prior to immobilization, 90-92% remained viable after formation and extensive curing of gels for cell-gel mixtures prepared at 45 degrees C . Attempts at prolonged storage of cell-laden gel beads at 0 degrees C as stock cultures of immobilized B . firmus were unsuccessful due to a significant decline in cell viability during such storage . On the basis of the cell leakage studies, the average pore sizes of 2%, 3%, 4%, and 5% gels were deduced to increase in the following order of carrageenan content (w/v): 4%, 3%, 2%, and 5% . Commensurate with the decrease in the average pore size (or the increased tightness of the gels) with increasing carrageenan content, both the extent of cell leakage and the extent of net cell penetration decreased with increasing carrageenan content for the first three gels . Owing to non-uniform distribution of free space and much larger pores, the extent of net cell penetration in 5% carrageenan gels was considerably low, while the extent of cell leakage in 5% carrageenan gels was an order of magnitude greater than the extents of cell leakage in the other three gels. Nature, 1991 Oct 31, 353(6347), 815 - 21 Crystal structure of insecticidal delta-endotoxin from Bacillus thuringiensis at 2.5 A resolution; Li JD et al.; The structure of the delta-endotoxin from Bacillus thuringiensis subsp . tenebrionis that is specifically toxic to Coleoptera insects (beetle toxin) has been determined at 2.5 A resolution . It comprises three domains which are, from the N- to C-termini, a seven-helix bundle, a three-sheet domain, and a beta sandwich . The core of the molecule encompassing all the domain interfaces is built from conserved sequence segments of the active delta-endotoxins . Therefore the structure represents the general fold of this family of insecticidal proteins . The bundle of long, hydrophobic and amphipathic helices is equipped for pore formation in the insect membrane, and regions of the three-sheet domain are probably responsible for receptor binding. Proc Natl Acad Sci U S A, 1991 Oct 15, 88(20), 8930 - 3 Binding of Bacillus thuringiensis proteins to a laboratory-selected line of Heliothis virescens; MacIntosh SC et al.; A laboratory-selected colony of Heliothis virescens displaying a 20- to 70-fold level of resistance to Bacillus thuringiensis proteins was evaluated to identify mechanism(s) of resistance . Brush-border membrane vesicles were isolated from larval midgut epithelium from the susceptible and resistant strains of H . virescens . Two B . thuringiensis proteins, CryIA(b) and CryIA(c), were iodinated and shown to specifically bind to brush-border membrane vesicles of both insect strains . Multiple changes in the receptor-binding parameters were seen in the resistant strain as compared with the susceptible strain . A 2- to 4-fold reduction in binding affinity was accompanied by a 4- to 6-fold increase in binding-site concentration for both proteins . Although these two B . thuringiensis proteins competed for the same high-affinity binding site, competition experiments revealed different receptor specificity toward these proteins in the resistant H . virescens line . The H . virescens strains were not sensitive to a coleopteran-active protein, CryIIIA, nor did these proteins compete with the CryIA proteins for binding . Complexity of the mechanism of resistance is consistent with the complex mode of action of B . thuringiensis proteins. J Immunol, 1991 Oct 15, 147(8), 2802 - 8 Cellular models of macrophage tumoricidal effector mechanisms in vitro . Characterization of cytolytic responses to tumor necrosis factor and nitric oxide pathways in vitro; Klostergaard J et al.; The recently described L-arginine-dependent nitric oxide (NO) pathway has been proposed to interact synergistically with the TNF pathway in murine macrophage-mediated tumor cytotoxicity in vitro . We have employed an experimental construct in which these two pathways were independently expressed by two different effector cell populations . The TNF-dependent pathway was committed by murine 3T3 cells transfected with the cDNA encoding human pro-TNF . The NO pathway was executed by the murine EMT-6 mammary adenocarcinoma cell line treated with murine rIFN-gamma and LPS . Controls for the TNF pathway committed by the transfectant included lysis of the TNF-sensitive murine L929 cell in coculture, secretion of TNF, and absence of nitrite synthesis . For the NO pathway controls included lysis of the murine P815 mastocytoma cocultured with activated EMT-6 cells that had been pretreated with murine rIFN-gamma and LPS, production of nitrite by this activated effector cell, and an absence of TNF secretion . The target cell panel included L929, EMT-6, P815, and murine B16 melanoma and TU-5 sarcoma cell lines . All targets on this panel were susceptible to lysis by LPS-triggered murine bacillus Calmette-Guerin-activated macrophages . The 3T3 transfectant caused significant lysis of cocultured L929 and TU-5 targets . The EMT-6 effector cell only caused significant lysis of the P815 target . When both effector cells were cocultured with these target cells, lysis of the P815 target was observed to be additive or superadditive; however, for all the other targets, cytotoxicity was comparable with or subadditive compared with that seen with the 3T3 transfectant effector cell alone . Thus, these two pathways do not appear to account for the broad, potent tumoricidal activity observed for activated macrophages in vitro. FEMS Microbiol Lett, 1991 Oct 15, 67(3), 273 - 6 Cytotoxicity of a cloned Bacillus thuringiensis subsp . israelensis CryIVB toxin to an Aedes aegypti cell line; Angsuthanasombat C et al.; A cloned CryIVB toxin was purified from a cured strain of Bacillus thuringiensis (BT) containing the cryIVB gene on the recombinant plasmid Cam135 . Solubilized protoxin was treated with Aedes gut extract or trypsin for varying times and tested for toxicity in vitro on three dipteran and one lepidopteran cell line . Treatment with the Aedes extract but not trypsin, produced an active toxin which lysed only Aedes aegypti cells out of those tested . This activation was time-dependent reaching a maximum after 6 h . Both the Aedes extract-treated and trypsin-treated toxin killed A . aegypti larvae, but this toxicity declined rapidly with increasing time of exposure to the proteolytic preparations. J Immunol, 1991 Oct 15, 147(8), 2706 - 12 Murine T cell-stimulatory peptides from the 19-kDa antigen of Mycobacterium tuberculosis . Epitope-restricted homology with the 28-kDa protein of Mycobacterium leprae; Harris DP et al.; Fifteen overlapping synthetic peptides, spanning the entire amino acid sequence of the Mycobacterium tuberculosis 19-kDa protein, were used to identify epitopes recognized by murine T cells . Five of the 15 peptides tested were able to elicit in vitro lymph node T cell proliferative responses in C57BL/10 mice primed by footpad inoculation with homologous peptide . Analysis in congenic strains of mice revealed H-2 restriction in the response to four peptides . However, one peptide, 19.7 (residues 61 to 80), induced T cell responses in all four haplotypes tested . This peptide was also unique in being able to stimulate lymph node cells from C57BL/10 mice immunized with recombinant 19-kDa protein, killed M . tuberculosis, or live bacillus Calmette Guerin infection . T cell lines specific for peptide 19.7 were of the CD4 phenotype . Significantly, sequence analysis revealed that residues 61 to 80 of the 19-kDa protein exhibited considerable homology with a single 20-amino acid sequence (residues 120 to 140), but not with any other region of the 28-kDa protein expressed in Mycobacterium leprae . This finding is the first evidence of epitope-restricted homology between otherwise structurally unrelated microbial Ag. Eur J Biochem, 1991 Oct 15, 201(2), 467 - 73 The electrochemical proton potential of Bacillus alcalophilus; Hoffmann A et al.; Bacillus alcalophilus strain ATCC 27647 showed usual growth characteristics, when inoculated at pH 10.4 . The cells entered the logarithmic phase at pH 10.3, and as growth continued, the pH dropped further to a value of 8.8 in the stationary phase . B . alcalophilus strain DSM 485 showed comparable growth only in the initial phase after the addition to fresh medium . The small initial growth period was succeeded by a long lag phase, where the pH continuously dropped . The cells resumed growth after the pH was about 10.0 and continued to grow accompanied by a further decrease of external pH . The bioenergetic parameters measured in the initial growth phase of the two strains at high pH (10.1-10.3) were nearly the same, i.e . delta pH = +97 to +110 mV, delta psi = -206 to -213 mV and delta microH+ = -109 to -103 mV . The inverted proton gradient of about 1.7-1.9 at high pH decreased, as the external pH dropped during growth . This led to an increase of the proton motive force (delta microH+), although the membrane potential (delta psi) also declined . The ATP/ADP ratio of strain DSM 485 was high (4.5-5.5) at fast growth during the initial and second growth period . The ratio declined to about 1.5 at the end of the lag phase . At the initial growth phase and at the end of the lag phase, the delta microH+ was, however, the same (approximately -106 mV) and considerably lower than in the middle of the second growth period (approximately -140 mV) . Fast growth, therefore, correlates with a high ATP/ADP ratio but not necessarily with a high delta microH+ . Addition of gramicidin or carbonylcyanide m-chlorophenylhydrazone stopped growth of B . alcalophilus strain DSM 485 at pH 10.3 or 9.5 and gramicidin immediately decreased the internal ATP/ADP ratio from 4.5 to 1.2 at pH 10.3. J Biol Chem, 1991 Oct 5, 266(28), 18567 - 72 Characterization of semi-uncoupled hybrid Escherichia coli-Bacillus megaterium F1F0 proton-translocating ATPases; Scarpetta MA et al.; Cloned atp genes for the proton-translocating ATPase of the obligate aerobe Bacillus megaterium have been demonstrated to be capable of complementing Escherichia coli ATPase (unc) mutants (Hawthorne, C . A., and Brusilow, W . S . A . (1986) J . Biol . Chem . 261, 5245-5248) . To determine the minimum subunit requirements for cross-species complementation, we constructed all combinations of B . megaterium atpA, G, D, and C genes (coding for the alpha, gamma, beta, and epsilon subunits, respectively) and tested their abilities to complement two uncA (alpha subunit) and two uncD (beta subunit) mutants of E . coli . The results indicated that complementation of either uncD mutant required atpD (beta) only . Complementation of one of the uncA (alpha) mutants required atpA, G, and D (alpha, gamma, and beta) and possibly atpE (epsilon) as well . The other uncA mutant was not complemented by any combination of B . megaterium ATPase genes . Complementation of a beta mutant by atpD (beta) or atpD and C (beta epsilon) produced cells which could grow aerobically on a nonfermentable carbon source (succinate) but not anaerobically on rich medium containing glucose . These E . coli therefore had become obligate aerobes . The ability to grow anaerobically could be restored to the mutant complemented by atpD alone by growth at pH 7.5 or pH 8 in the presence of 0.1 M potassium. Biochem J, 1991 Oct 1, 279 ( Pt 1), 213 - 21 Mechanism of acyl transfer by the class A serine beta-lactamase of Streptomyces albus G; Lamotte-Brasseur J et al.; Optimization by energy minimization of stable complexes occurring along the pathway of hydrolysis of benzylpenicillin and cephalosporin C by the Streptomyces albus G beta-lactamase has highlighted a proton shuttle that may explain the catalytic mechanism of the beta-lactamases of class A . Five residues, S70, S130, N132, T235 and A237, are involved in ligand binding . The gamma-OH group of T235 and, in the case of benzylpenicillin, the gamma-OH group of S130 interact with the carboxylate group, on one side of the ligand molecule . The side-chain NH2 group of N132 and the carbonyl backbone of A237 interact with the exocyclic CONH amide bond, on the other side of the ligand . The backbone NH groups of S70 and A237 polarize the carbonyl group of the scissile beta-lactam amide bond . Four residues, S70, K73, S130 and E166, and two water molecules, W1 and W2, perform hydrolysis of the bound beta-lactam compound . E166, via W1, abstracts the proton from the gamma-OH group of S70 . While losing its proton, the O-gamma atom of S70 attacks the carbonyl carbon atom of the beta-lactam ring and, concomitantly, the proton is delivered back to the adjacent nitrogen atom via W2, K73 and S130, thus achieving formation of the acyl-enzyme . Subsequently, E166 abstracts a proton from W1 . While losing its proton, W1 attacks the carbonyl carbon atom of the S70 ester-linked acyl-enzyme and, concomitantly, re-entry of a water molecule W'1 replacing W1 allows E166 to deliver the proton back to the same carbonyl carbon atom, thus achieving hydrolysis of the beta-lactam compound and enzyme recovery . The model well explains the differences found in the kcat . values for hydrolysis of benzylpenicillin and cephalosporin C by the Streptomyces albus G beta-lactamase . It also explains the effects caused by site-directed mutagenesis of the Bacillus cereus beta-lactamase I {Gibson, Christensen & Waley (1990) Biochem J . 272, 613-619}. Biochem J, 1991 Oct 1, 279 ( Pt 1), 111 - 4 Interaction of beta-lactamases I and II from Bacillus cereus with semisynthetic cephamycins . Kinetic studies; Martin Villacorta J et al.; The influence of C-6 alpha- or C-7 alpha-methoxylation of the beta-lactam ring in the catalytic action of class A and B beta-lactamases has been investigated . For this purpose the kinetic behaviour of beta-lactamases I (class A) and II (class B) from Bacillus cereus was analysed by using several cephamycins, moxalactam, temocillin and related antibiotics . These compounds behaved as poor substrates for beta-lactamase II, with high Km values and very low catalytic efficiencies . In the case of beta-lactamase I, the substitution of a methoxy group for a H atom at C-7 alpha or C-6 alpha decreased the affinity of the substrates for the enzyme . Furthermore, the acylation of cephamycins was completely blocked, whereas that of penicillins was slowed down by a factor of 10(4)-10(5), acylation being the rate-determining step of the process. J Bacteriol, 1991 Oct, 173(19), 6147 - 52 Analysis of the active center of Bacillus stearothermophilus neopullulanase; Kuriki T et al.; The active center of the neopullulanase from Bacillus stearothermophilus was analyzed by means of site-directed mutagenesis . The amino acid residues located in the active center of the neopullulanase were tentatively identified according to a molecular model of Taka-amylase A and homology analysis of the amino acid sequences of neopullulanse, Taka-amylase A, and other amylolytic enzymes . When amino acid residues Glu and Asp, corresponding to the putative catalytic sites, were replaced by the oppositely charged (His) or noncharged (Gln or Asn) amino acid residue, neopullulanase activities toward alpha-(1----4)- and alpha-(1----6)-glucosidic linkages disappeared . When the amino acids corresponding to the putative substrate-binding sites were replaced, the specificities of the mutated neopullulanases toward alpha-(1----4)- and alpha-(1----6)-glucosidic linkages were obviously different from that of the wild-type enzyme . This finding proves that one active center of neopullulanase participated in the dual activity toward alpha-(1----4)- and alpha-(1----6)-glucosidic linkages . Pullulan is a linear glucan of maltotriosyl units linked through alpha-(1----6)-glucosidic linkages . The production ratio of panose from pullulan was significantly increased by using the mutated neopullulanase which exhibited higher specificity toward the alpha-(1----4)-glucosidic linkage . In contrast, the production ratio of panose was obviously decreased by using the mutated neopullulanse which exhibited higher specificity toward the alpha-(1----6)-glucosidic linkage. Eur J Biochem, 1991 Oct 1, 201(1), 203 - 9 Sequence-specific 1H-NMR assignments and secondary structure of the lipoyl domain of the Bacillus stearothermophilus pyruvate dehydrogenase multienzyme complex; Dardel F et al.; The lipoyl domain (residues 1-85) of the lipoate-acetyltransferase polypeptide chain of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus has been subjected to detailed structural analysis by means of two-dimensional (2D) 1H-NMR spectroscopy at 400 MHz . Sequence-specific proton resonance assignments were made, but at this field strength not all of the side-chain protons could be assigned, especially from complex spin systems like those of leucine, proline and lysine residues . Measurement of short-range interproton distances identified two extensive regions of beta-sheet, each containing four anti-parallel peptide strands . The lipoyl-lysine residue (Lys42) is located in a tight turn at a corner of one sheet, the N-terminal and C-terminal residues of the domain are close together in two adjacent beta-strands in the other . The lipoylated and unlipoylated forms of the domain have almost identical spectra, indicating that there is little, if any, conformational change in the protein as a result of the post-translational modification. J Urol, 1991 Oct, 146(4), 1164 - 7 Treatment of murine transitional cell carcinoma with intralesional interleukin 2 and murine interferon gamma; Sosnowski JT et al.; The antitumor effect of intralesionally administered recombinant interleukin-2, alone or in combination with recombinant interferon gamma was studied in murine transitional cell carcinoma, MBT2 . In the initial prophylactic model treatment was started at day one at the site of tumor inoculation . Maximal and significant reduction in tumor volume occurred in groups receiving 4,000 units of recombinant interleukin 2 and 10(7) colony forming units Bacillus Calmette Guerin (p less than 0.00001 vs saline control) . In the same experiment, a reduction in tumor incidence and increase in survival occurred in groups receiving 4,000 units of recombinant interleukin 2, 1,000 units of recombinant interleukin 2 plus 2,000 units of recombinant interferon gamma, as well as 10(7) colony forming units Bacillus Calmette Guerin relative to saline control (p less than 0.005) . The dose-response effect of recombinant interleukin 2 alone was also tested in a model of an established transitional cell carcinoma . Intralesional injection treatments were initiated after tumors were palpable . Reduction in tumor volume was observed in the group receiving 8,000 units of recombinant interleukin 2 (p = 0.01 vs saline control), but no significant advantage in survival was noted. J Urol, 1991 Oct, 146(4), 1118 - 9 A case of granulomatous hepatitis after intravesical bacillus Calmette-Guerin administration; Graziano DA et al.; A 61-year-old man received intravesical bacillus Calmette-Guerin (BCG) as treatment for superficial bladder carcinoma . High spiking relapsing fevers began 39 days after the initial treatment . A liver biopsy revealed noncaseating granuloma . Deoxyribonucleic acid hybridization of the bone marrow was positive for Mycobacterium tuberculosis complex . Pressure exerted to instill the BCG may have favored dissemination. Indian J Exp Biol, 1991 Oct, 29(10), 953 - 7 An enzyme linked immunosorbent assay for monitoring Bacillus sphaericus toxin; Anandkumar K et al.; Larvicidal proteins of B . sphaericus H5a5b (strain VCRC B42), purified by ion-exchange chromatography were used to raise antibodies in rabbits . The antibodies were specific in reacting to alkali-solubilized fractions from whole cells of toxic strains only . Ouchterlony immunodiffusion showed homology in toxin structure between strains of different serotype . A sandwich ELISA using avidin-biotin amplification was standardized . The lowest detectable limit was 6.25 ng/ml . Near linear quantitative binding of the antigen was found in the range 25-200 ng/ml . The growth, toxin level and LC50 values during various stages of fermentation of B . sphaericus strains 1593 and B42 were compared . There was significant correlation between LC50 values and toxin levels as measured by ELISA. Antonie Van Leeuwenhoek, 1991 Oct-Nov, 60(3-4), 355 - 71 Formation of fermentation products and extracellular protease during anaerobic growth of Bacillus licheniformis in chemostat and batch-culture; Bulthuis BA et al.; For a relaxed (rel-), protease producing (A-type) and a stringent (rel+), not-protease producing (B-type) variant of Bacillus licheniformis we determined fermentation patterns and products, growth parameters and alkaline protease-production (if any) in anaerobic, glucose-grown chemostats and batch-cultures . Glucose is dissimilated via glycolysis and oxidative pentose phosphate pathway simultaneously; the relative share of these two routes depends on growth phase (in batch) and specific growth rate (in chemostat) . Predominant products are lactate, glycerol and acetaldehyde for A-type batches and acetaldehyde, ethanol, acetate and lactate for B-type batches . Both types show a considerable acetaldehyde production . In chemostat cultures, the fermentation products resemble those in batch-culture . From the anaerobic batches and chemostats, we conclude that the A-type (with low ATP-yield) will have a YATPmax of probably 12.9 g/mol and the B-type (with high ATP-yield) a YATPmax of about 10.1 g/mol . For batch-cultures, both types have about the same, high Yglucose (12 g/mol) . So, the slow-growing A-type has a relatively high efficiency of anaerobic growth (i.e . an efficient use of ATP) and the fast-growing B-type a relatively low efficiency of anaerobic growth . In aerobic batch-cultures, we found 48, respectively 41% glucose-carbon conversion into mainly glycerol and pyruvate, respectively acetate as overflow metabolites in the A- and B-type . In both aerobic and anaerobic batch-cultures of the A-type, protease is produced predominantly in the logarithmic and early stationary phase, while a low but steady production is maintained in the stationary phase . Protease production occurs via de novo synthesis; up to 10% of the total protease in a culture is present in a cell-associated form . Although anaerobic protease production (expressed as protease per amount of biomass) is much higher than for aerobic conditions, specific rates of production are in the same range as for aerobic conditions while, most important, the substrate costs of anaerobic production are very much higher than for aerobic conditions. Arch Esp Urol, 1991 Oct, 44(8), 1025 - 8 Immunoprophylaxis of superficial bladder cancer: a prospective and randomized comparison of oral versus intravesical Bacillus Calmette-Guerin; Netto Junior NR et al.; A total of 84 patients with superficial transitional cell carcinoma underwent transurethral resection of bladder tumor . All patients had stage pTa or pT1 transitional cell carcinoma or carcinoma in situ without other concurrent malignancies . The patients were assigned to three treatment groups: I . Control group-transurethral resection (TUR-BT) discontinued within the study . II . Oral BCG group-TUR-BT plus BCG (Moreau) . III . Intravesical BCG group-TUR-BT plus BCG . Of 9 patients in the control group, 8 (89%) experienced tumor recurrence during a mean follow-up of 20 months . Of the 33 patients in the oral BCG group, 13 patients (39.3%) had recurrence during a mean follow-up of 39 months . Of the 42 patients in the intravesical group, 8 patients (19%) had recurrence in a 30-month mean follow-up . The incidence of complications was higher in the intravesical (33.4%) than in the oral BCG group (24.2%) . These results showed that intravesical BCG is a more effective immunotherapy; however, oral BCG can be utilized in patients who do not accept intravesical BCG administration. Xenobiotica, 1991 Oct, 21(10), 1311 - 23 Hydroxylation and glucoside conjugation in the microbial metabolism of the diterpene sclareol; Kouzi SA et al.; 1 . The microbial metabolism of sclareol (1), a labdane diterpene ditertiary alcohol, was studied . Preliminary screening identified a number of microorganisms capable of metabolizing sclareol . 2 . Preparative scale fermentation of growing cultures of Bacillus cereus UI-1477 resulted in the production of seven metabolites which have been characterized as 3 beta-hydroxysclareol (2), 2 alpha-hydroxysclareol (3), 18-hydroxysclareol (4), 2 alpha,18-dihydroxysclareol (6), 8 alpha,13 beta-dihydroxy-labd-14-en-3 beta-O-beta-D-glucoside (5), 8 alpha,13 beta-dihydroxy-labd-14-en-18-O-beta-D-glucoside (7), and 8 alpha,13 beta-dihydroxy-labd-14-en-2 alpha-O-beta-D-glucoside (10) by chemical, enzymic, and spectral data, especially 2D-n.m.r . techniques and thermospray liquid chromatography-mass spectrometry analysis. Acta Crystallogr B, 1991 Oct 1, 47 ( Pt 5), 707 - 30 Complex between the subtilisin from a mesophilic bacterium and the leech inhibitor eglin-C; Dauter Z et al.; The alkaline proteinase from the mesophilic bacterium Bacillus mesentericus has been crystallized in a 1:1 complex with the inhibitor eglin-C from the medical leech . The crystals have cell dimensions of a = 43.0, b = 71.9, c = 48.3 A and beta = 110.0 degrees and are in the space group P2(1) . Three-dimensional data to 2.0 A have been recorded on film from a single crystal . The orientation and position of the complex in the unit cell have been established using the refined coordinates of subtilisin Carlsberg and of eglin-C as independent models . The structure of the complex has been refined by restrained least-squares minimization . The crystallographic R factor (= sigma{magnitude of Fo - magnitude of Fc{/sigma magnitude of Fo) is 15.1% including two Ca2+ ions and 312 water molecules . The structure is discussed in terms of its physicochemical properties in solution and its relation to other Bacillus subtilisins. Anal Biochem, 1991 Oct, 198(1), 10 - 4 A fluorescent substrate for the continuous assay of phosphatidylinositol-specific phospholipase C: synthesis and application of 2-naphthyl myo-inositol-1-phosphate; Shashidhar MS et al.; A fluorescent water-soluble substrate for phosphatidylinositol-specific phospholipase C was synthesized . The diacylglycerol moiety of the natural substrate, phosphatidylinositol, was replaced by the fluorescent moiety, 2-naphthol, resulting in the synthetic substrate, racemic 2-naphthyl myo-inositol-1-phosphate . The synthetic substrate provided a continuous fluorometric assay for the phosphatidylinositol-specific phospholipase C from Bacillus cereus . Initial rates of the cleavage of the 2-naphthyl substrate by the phospholipase measured by fluorometry were linear with time and the amount of enzyme added . The specific enzyme activity at pH 8.5 and 25 degrees C was about 0.04 mumol/min mg protein at an initial substrate concentration of 0.8 mM . 31P NMR experiments suggest that, as with phosphatidylinositol itself, cleavage of the fluorescent substrate proceeds in two steps via a myo-inositol-1,2-cyclic phosphate intermediate, and that only the D-isomer is a substrate for the B . cereus phospholipase . The synthetic substrate was stable during long-term storage as a solid in the dark at -20 degrees C . It was also stable for several weeks when stored in the dark frozen in aqueous solution near neutral pH. FEMS Microbiol Lett, 1991 Oct 1, 67(2), 187 - 91 Taxonomic considerations of Bartonella bacilliformis based on phylogenetic and phenotypic characteristics; Birtles RJ et al.; The 16S-rRNA gene of Bartonella bacilliformis was amplified using the polymerase-chain reaction (PCR) . The amplification product was sequenced using a linear-PCR procedure and compared with other published 16S-rRNA sequences . The results of this analysis placed B . bacilliformis in the alpha subgroup of the proteobacteria, and more specifically demonstrated its close phylogenetic relationship to Rochalimaea quintana . This relationship is supported by similarities in the size and mean base composition of the genomes of the two species, and by shared phenotypic characteristics. Can J Microbiol, 1991 Oct, 37(10), 775 - 9 Reversibility of oxygen switch-off effect on Bacillus polymyxa nitrogenase; Holzmann HP et al.; The objective of this study was to analyse in vivo the effect of oxygen on the nitrogenase of Bacillus polymyxa . The culture technique employed in this study prevented spore formation by B . polymyxa during the entire period of exposure to acetylene . Under these conditions the acetylene-reduction assay allowed quantification of nitrogenase activity over long incubation periods (44 h) . Nitrogenase activity was highest in cells harvested in the late logarithmic phase . At PO2 of 0.19 and 0.37 kPa, acetylene reduction was inhibited by 80 and 100%, respectively . This switch-off effect could be reversed through oxygen exhaustion, either by flushing the culture with N2 or by cellular respiration, suggesting a respiratory protection mechanism for the nitrogenase complex in B . polymyxa . Oxygen consumption measured by a closed-chamber respirometer showed a linear increase up to a PO2 of 0.2 kPa . Above 0.3 kPa a saturation in oxygen consumption was observed . Exposure to high oxygen pressures resulted in an irreversible loss of nitrogenase activity . The oxygen inhibition pattern was shown to be similar to that in other microaerophilic and anaerobic nitrogen-fixing microorganisms. J Gen Microbiol, 1991 Oct, 137 ( Pt 10), 2375 - 9 Molar growth yields and bioenergetic parameters of extremely alkaliphilic Bacillus species in batch cultures, and growth in a chemostat at pH 10.5; Guffanti AA et al.; Alkaliphilic Bacillus species that grow at pH 10.5 must cope with a low protonmotive force (-50 mV) due to a reversed transmembrane pH gradient at least 2 pH units more acid inside . Here we demonstrate that strictly alkaliphilic B . firmus RAB and two strains of B . alcalophilus (ATCC 27467 and DSM 485) grow exponentially in batch cultures with a doubling time of less than 1 h in 100 mM buffered medium, while the actual medium pH remains above 10.2 . The ATCC strain continued to grow rapidly for at least 7 h, but the growth rate of the DSM strain declined dramatically after 3 h . However, both the B . alcalophilus strains, B . firmus RAB and facultatively alkaliphilic B . firmus OF4 were readily maintained for at least 24 h between pH 10.4 and 10.6 in a chemostat where nutrients were constantly replenished . A critical nutrient may be limiting in batch cultures of the DSM strain of B . alcalophilus . The facultative alkaliphile grew equally well in batch cultures at an initial pH of 7.5 or 10.5 . Its molar growth yield (23 mg dry wt mmol-1) on malate (Ymal) was the same at the two pH values and was comparable to Ymal for B . subtilis grown at neutral pH . B . firmus RAB and B . alcalophilus ATCC 27467 grown at pH 10.5 also showed Ymal values at least as high as the neutralphile, indicating efficient use of the energy source even at low protonmotive force . Moreover, the phosphorylation potential of B . firmus OF4 grown at pH 7.5 (45.2 kJ mol-1) or pH 10.5 (46 kJ mol-1) was in a conventional range for bacteria. Chirurg, 1991 Oct, 62(10), 732 - 8 {Helicobacter pylori colonization in surgical patients}; Ludtke FE et al.; The prevalence of Helicobacter pylori (HP) was examined in 387 patients undergoing endoscopy of the upper gastrointestinal tract . Of central interest was the question to which extent surgical intervention influences the colonisation of the gastric mucosa with HP . The bacillic status was appraised using double microbiological examinations, histological determination and the CLO-test . In 229 patients a 13C-urea-breath test was also carried out (sensitivity 98%) . HP could be detected in 90% of all patients presenting with duodenal ulcers as well as in 70% of patients with gastric ulcers, whereas in those patients in whom a lesion of the upper gastrointestinal tract could be excluded through endoscopy . HP was found in only 27% . The prevalence of HP did not increase with age . In patients having undergone distal gastric resection due to gastric ulcers, HP was only rarely found (19%) in the mucosa in the vicinity of the anastomosis following removal of the apparently pathogenetically important antrum mucosa . There was no association between anastomosis ulcers and bacillic colonisation . Following selective proximal vagotomy in patients with duodenal ulcers, HP was found in 80% of all cases . In these patients there was also no association between recurrent ulceration and a HP-positive status . Our results describe the postoperative HP-status after different surgical procedures of ulcer therapy: whereas a distal gastric resection removes the antrum mucosa, which provides the necessary environmental milieu, the HP-colonisation rate after selective proximal vagotomy is similar to that in non-operated ulcer disease. Appl Environ Microbiol, 1991 Oct, 57(10), 2816 - 20 Identification of putative insect brush border membrane-binding molecules specific to Bacillus thuringiensis delta-endotoxin by protein blot analysis; Garczynski SF et al.; Binding sites for insecticidal toxins of Bacillus thuringiensis are located in the brush border membranes of insect midguts . Two approaches were used to investigate the interactions of B . thuringiensis subsp . kurstaki HD-73 CryIA(c) toxin with brush border membrane vesicles from sensitive and naturally resistant insects: 125I-toxin-vesicle binding assays and protein blots probed with 125I-CryIA(c) toxin . In bioassays, Manduca sexta and Heliothis virescens larvae were highly sensitive, Helicoverpa zea larvae were moderately sensitive, and Spodoptera frugiperda larvae were resistant to CryIA(c) toxin . Studies of binding of 125I-CryIA(c) toxin to brush border membrane vesicles from the larval midguts revealed that all insects tested had high-affinity, saturable binding sites . Significantly, S . frugiperda larvae bind but are not killed by CryIA(c) toxin . Labeled CryIA(c) toxin incubated with protein blots identifies a major binding molecule of 120 kDa for M . sexta and 148 kDa for S . frugiperda . H . virescens and H . zea are more complex, containing 155-, 120-, 103-, 90-, and 63-kDa proteins as putative toxin-binding molecules . H . virescens also contains a minor toxin-binding protein of 81 kDa . These experiments provide information that can be applied toward a more detailed characterization of B . thuringiensis toxin-binding proteins. Appl Environ Microbiol, 1991 Oct, 57(10), 2783 - 9 Tributyltin-resistant bacteria from estuarine and freshwater sediments; Wuertz S et al.; Resistance to tributyltin (TBT) was examined in populations from TBT-polluted sediments and nonpolluted sediments from an estuary and from fresh water as well as in pure cultures isolated from those sediments . The 50% effective concentrations (EC50s) for populations were higher at a TBT-polluted freshwater site than at a site without TBT, suggesting that TBT selected for a TBT-resistant population . In contrast, EC50s were significantly lower for populations from a TBT-contaminated estuarine site than for those from a site without TBT, suggesting that other factors in addition to TBT determine whether populations become resistant . EC50s for populations from TBT-contaminated freshwater sediments were nearly 30 times higher than those for populations from TBT-contaminated estuarine sediments . We defined a TBT-resistant bacterium as one which grows on trypticase soy agar containing 8.4 microM TBT, a concentration which prevented the growth of 90% of the culturable bacteria from these sediments . The toxicity of TBT in laboratory media was influenced markedly by the composition of the medium and whether it was liquid or solid . Ten TBT-resistant isolates from estuarine sediments and 19 from freshwater sediments were identified to the genus level . Two isolates, each a Bacillus sp., may be the first gram-positive bacteria isolated from fresh water in the presence of a high concentration of TBT . There was a high incidence of resistance to heavy metals: metal resistance indices were 0.76 for estuarine isolates and 0.68 for freshwater isolates.(ABSTRACT TRUNCATED AT 250 WORDS) Int J Syst Bacteriol, 1991 Oct, 41(4), 510 - 5 Bacillus brevis Migula 1900 taxonomy: reassociation and base composition of DNA; Nakamura LK; Of 87 strains previously identified as Bacillus brevis Migula 1900, 58 had G + C contents of 47.0 to 51.9 mol%, a range that included the G + C content (48.7 mol%) of the type strain . The G + C contents for three other groups consisting of 5, 7, and 17 strains were 37.0 to 41.9, 42.0 to 46.9, and 52.0 mol% or higher, respectively . DNA reassociation studies showed that 25 of the 58 strains with G + C contents of 47.0 to 51.9 mol% were closely related genetically to the type strain and to each other . For the most part, this genetically related group was phenotypically homogeneous; variations in the fermentation of mannitol and mannose were observed . My results strongly suggest that many of the strains were misclassified as B . brevis . Consequently, much of the phenotypic heterogeneity of the species B . brevis Migula 1900 is not due to variations exhibited by genetically related organisms, but is the result of variability introduced by the presence of genetically unrelated strains. Rev Saude Publica, 1991 Oct, 25(5), 367 - 70 {Susceptibility of populations of Simulium (Chirostilbia) pertinax Kollar, 1832 (Culicomorpha, Simuliidae) to eemephos and to Bacillus thuringiensis var . israelensis-based formulation}; de Andrade CF et al.; The use of wooden troughs on stream beds, artificially colonized by blackfly larvae, is proposed for larvicide evaluations . Mortality was recorded 3 or 4 hours after treatment . Larval susceptibility was also evaluated utilizing the LT50 criterion . In there field assays Simulium (C.) pertinax populations from the litoral of S . Paulo and Rio de Janeiro States were shown to be resistant to temephos, even when subjected to high concentrations . Vectobac 12 AS, a Bacillus thuringiensis var . israelensis product, was shown to be more potent against late instar larvae and efficient in concentrations higher than 7,200 ITU/l (10 min) . The LT50 to 3,744 ITU/l (10 min) was calculated as 70.9 min. Rev Latinoam Microbiol, 1991 Oct-Dec, 33(4), 279 - 92 {Bacillus thuringiensis: biological characteristics and production perspectives}; Rodriguez Monroy M et al.; In this paper the potential activity of Bacillus thuringiensis as insecticide is reviewed . It is presented recent knowledges of its metabolism, production system and the rinetic constants already reported finally the possibilities for production in batches or continuous culture is discussed in order to improve the productivity and economical aspects. Indian J Biochem Biophys, 1991 Oct-Dec, 28(5-6), 472 - 5 Isolation of Gln- mutants through transposon, Tn917, mutagenesis in Bacillus brevis; Mishra HS et al.; Transposon, Tn917, carried on pTV1 plasmid has been used successfully to mutagenise Bacillus brevis . The transposon showed preference for insertion at an "aro" site . A second insertional event after elimination of the preferred site with ethidium bromide/acridine orange treatment has permitted isolation of Gln- mutants in B . brevis. J Bacteriol, 1991 Oct, 173(20), 6635 - 8 Cloning and analysis of delta-endotoxin genes from Bacillus thuringiensis subsp . alesti; Lee CS et al.; Bacillus thuringiensis subsp . alesti produced only CryIA(b)-type protoxins, and three cryIA(b) genes were cloned . One was cryptic because of an alteration near the 5' end, and the other two were very similar to each other . The protoxin encoded by one of the latter genes differed from other CryIA(b) protoxins in its greater stability and relative toxicity for two members of the order Lepidoptera. Appl Microbiol Biotechnol, 1991 Oct, 36(1), 5 - 13 Biosurfactants from Bacillus licheniformis: structural analysis and characterization; Jenny K et al.; The surface-active compounds of the strain Bacillus licheniformis were isolated and their structure was elucidated . The high surface-active capacity of the crude extract was basically due to traces of long-chain saturated fatty acids, especially of palmitic and stearic acids, to a mixture of small amounts of hydrocarbons with chain lengths of 20 and 22 carbons, and mainly to appreciable amounts of four slightly different lipopeptides . The lipopeptides were found to be a mixture of four closely related compounds . The lipophilic part consisting of i-, n-C14 or i-, ai-C15 beta-OH fatty acids was linked to the hydrophilic peptide moiety, which contained seven amino acids (Glu, Asp, Val, three Leu and Ile) by a lactone linkage . Fifteen milligrams per litre of the purified lipopeptide product decreased the surface tension of water from 72 mN m-1 to 27 mN m-1, characterizing the product as a powerful surface-active agent that compares favourably to other (bio)surfactants . Antibiotic activity was demonstrated against bacteria and yeasts. J Biol Chem, 1991 Sep 25, 266(27), 17954 - 8 Functional domains of Bacillus thuringiensis insecticidal crystal proteins . Refinement of Heliothis virescens and Trichoplusia ni specificity domains on CryIA(c); Ge AZ et al.; Insecticidal crystal proteins (delta-endotoxins), CryIA(a) and CryIA(c), from Bacillus thuringiensis are 82% homologous . Despite this homology, CryIA(c) was determined to have 10-fold more insecticidal activity toward Heliothis virescens and Trichoplusia ni than CryIA(a) . Reciprocal recombinations between these two genes were performed by the homolog-scanning technique . The resultant mutants had different segments of their primary sequences exchanged . Bioassays with toxin proteins from these mutants revealed that amino acids 335-450 on CryIA(c) are associated with the activity against T . ni, whereas amino acids 335-615 on the same toxin are required to exchange full H . virescens specificity . One chimeric protein toxin, involving residues 450-612 from CryIA(c), demonstrated 30 times more activity against H . virescens than the native parental toxin, indicating that this region plays an important role in H . virescens specificity . The structural integrity of mutant toxin proteins was assessed by treatment with bovine trypsin . All actively toxic proteins formed a 65-kDA trypsin-resistant active toxic core, similar to the parental CryIA(c) toxin, indicating that toxin protein structure was not altered significantly . Contrarily, certain inactive mutant proteins were susceptible to complete protease hydrolysis, indicating that their lack of toxicity may have been due to structural alterations. Biochemistry, 1991 Sep 24, 30(38), 9195 - 200 Binding of a chaperonin to the folding intermediates of lactate dehydrogenase; Badcoe IG et al.; When Bacillus stearothermophilus LDH dimer is incubated with increasing concentrations of the denaturant guanidinium chloride, three distinct unfolded states of the molecule are observed at equilibrium {Smith, C . J., et al . (1991) Biochemistry 30, 1028-1036} . The kinetics of LDH refolding are consistent with an unbranched progression through these states . The Escherichia coli chaperonin, GroEL, binds with high affinity to the completely denatured form and more weakly to the earliest folding intermediate, thus retarding the refolding process . A later structurally defined folding intermediate, corresponding to a molten globule form, is not bound by GroEL; neither is the inactive monomer . The complex between GroEL and denatured LDH is destabilized by the binding of magnesium/ATP (Mg/ATP) or by the nonhydrolyzable analogue adenylyl imidodiphosphate (AMP-PNP) . From our initial kinetic data, we propose that GroEL exists in two interconvertible forms, one of which is stabilized by the binding of Mg/ATP but associates weakly with the unfolded protein . The other is destabilized by Mg/ATP and associates strongly with unfolded LDH . The relevance of these findings to the role of GroEL in vivo is discussed. FEBS Lett, 1991 Sep 23, 290(1-2), 221 - 3 Overproduction, purification and crystallization of Bacillus cereus oligo-1,6-glucosidase; Watanabe K et al.; The gene coding for oligo-1,6-glucosidase from Bacillus cereus ATCC7064 has been overexpressed in Escherichia coli MV1184 cells under the control of the lac promoter in the genetically engineered plasmid pBCE4-2 . Oligo-1,6-glucosidase was purified in large quantities and was crystallized at 25 degrees C by using a hanging drop vapor diffusion method with 53% saturated ammonium sulfate . The crystals have the shape of hexagonal bipyramids and belong to the space group P6(2) or P6(4) with lattice constants of a = b = 106.1 A, c = 120.0 A and gamma = 120 degrees. FEBS Lett, 1991 Sep 23, 290(1-2), 13 - 6 Hydrolysis of branched cyclodextrins by a cyclodextrin-hydrolyzing enzyme from Bacillus sphaericus E-244; Oguma T et al.; The action of a cyclodextrin-hydrolyzing enzyme from Bacillus sphaericus E-244 on branched alpha- and beta-cyclodextrins was investigated . Glucosyl-alpha-cyclodextrin (6-O-alpha-D-glucosylcyclomaltohexaose) and maltosyl-alpha-cyclodextrin (6-O-alpha-D-maltosylcyclomaltohexaose) were hydrolyzed to 6(3)-O-alpha-D-glucosylmaltohexaose and 6(3)-O-alpha-D-maltosylmaltohexaose, respectively . Glucosyl-beta-cyclodextrin (6-O-alpha-D-glucosylcyclomaltoheptose) and maltosyl-beta-cyclodextrin (6-O-alpha-D-maltosyclomaltohepatose) were also mainly transformed to 6(4)-O-alpha-D-glucosylmaltoheptaose and 6(4)-O-alpha-D-maltosylmaltoheptaose, respectively . These results suggest that the cyclodextrin-hydrolyzing enzyme cleaves branched alpha- and beta-cyclodextrins at an alpha-1,4 linkage which is located furthest from the branching point on the cyclodextrin ring. Biochim Biophys Acta, 1991 Sep 20, 1079(3), 247 - 52 Duck liver malic enzyme: sequence of a tryptic peptide containing the cysteine residue labeled by the substrate analog bromopyruvate; Satterlee J et al.; Malic enzyme of duck liver is alkylated by bromopyruvate with half-of-the-sites stoichiometry, and with accompanying loss of oxidative decarboxylase and enhancement of pyruvate reductase activities as was previously shown for the pigeon enzyme (Hsu, R.Y . (1982) Mol . Cell . Biochem . 43, 3-26) . In the present work, the alkylated enzyme is shown to bind NADPH, but not L-malate in the presence of MnCl2, indicating impairment of the enzyme site for the substrate and/or divalent metal . The enzyme was differentially labeled by 3-bromo-1-{14C}-pyruvate and digested with TPCK-treated trypsin . Two peptides bearing the susceptible residue were purified by high-performance liquid chromatography and sequenced . Peptide II has the sequence of FMPIVYTPTVGLAXQQYGLAFR, corresponding to residues 86-107 (temporary numbering) of the duck enzyme; cysteine-99(x) is not detected, indicating that it is the target of modification by bromopyruvate . Peptide I is a truncated form of peptide II lacking five amino acid residues at the C-terminal . Cysteine-99 is conserved in malic enzymes from duck, rat, mouse, maize, human, Flaveria trinervia and Bacillus stearothermophilus. Biochem Biophys Res Commun, 1991 Sep 16, 179(2), 933 - 8 The toxic moiety of the Bacillus thuringiensis protoxin undergoes a conformational change upon activation; Choma CT et al.; Proteolytic processing of the 133-kDa crystal protein (protoxin) from Bacillus thuringiensis subsp . kurstaki yields a 67-kDa insecticidal toxin . Differential scanning calorimetry was used to investigate whether the toxic moiety in the protoxin molecule has the same conformation as activated toxin . Compared to protoxin, toxin gives rise to a more complex endotherm which extends over a 10 degrees C broader temperature range and contains a component occurring at a substantially higher temperature than any unfolding transition in the protoxin endotherm . It is concluded that the toxic moiety undergoes a conformational change upon activation in which the thermal stability of at least one of its domains is significantly increased. Biochem J, 1991 Sep 15, 278 ( Pt 3), 809 - 16 Evidence that gene G7a in the human major histocompatibility complex encodes valyl-tRNA synthetase; Hsieh SL et al.; At least 36 genes have now been located in a 680 kb segment of DNA between the class I and class II multigene families within the class III region of the human major histocompatibility complex on chromosome 6p21.3 . The complete nucleotide sequence of the 4.3 kb mRNA of one of these genes, G7a (or BAT6), has been determined from cDNA and genomic clones . The single-copy G7a gene encodes a 1265-amino-acid protein of molecular mass 140,457 Da . Comparison of the derived amino acid sequence of the G7a protein with the National Biomedical Research Foundation protein databases revealed 42% identity in a 250-amino-acid overlap with Bacillus stearothermophilus valyl-tRNA synthetase, 38.0% identity in a 993-amino-acid overlap with Escherichia coli valyl-tRNA synthetase (val RS), and 48.3% identity in a 1043-amino-acid overlap with Saccharomyces cerevisiae valyl-tRNA synthetase . The protein sequence of G7a contains two short consensus sequences, His-Ile-Gly-His and Lys-Met-Ser-Lys-Ser, which is the typical signature structure of class I tRNA synthetases and indicative of the presence of the Rossman fold . In addition, the molecular mass of the G7a protein is the same as that of other mammalian valyl-tRNA synthetases . These features and the high sequence identity with yeast valyl-tRNA synthetase strongly support the fact that the G7a gene, located within the major histocompatibility complex, encodes the human valyl-tRNA synthetase. J Urol, 1991 Sep, 146(3), 766 - 9; discussion 769-70 Bacillus Calmette-Guerin and interleukin-2 for treatment of superficial bladder cancer; Cockett AT et al.; A total of 22 patients with bladder cancer received bacillus Calmette-Guerin (BCG) and interleukin-2 . Significant bladder tumor remissions were noted in 15 of 17 patients (88%) . Of 5 patients with carcinoma in situ 1 was noncompliant and he died of carcinoma in situ . The other 4 patients are in remission . BCG alone was instilled in 22 additional patients with superficial bladder cancer . The remission rates were encouraging . Of the 22 patients 13 (59%) had remission of the bladder tumor . A half dose of BCG (60 mg.) is adequate when given weekly for 6 weeks . Maintenance therapy is important as noted in both of our clinical arms . BCG and interleukin-2 therapy results in a higher remission rate. FEBS Lett, 1991 Sep 9, 289(2), 148 - 50 Modification of alpha-amylase from Bacillus licheniformis by the polyaldehyde derived from beta-cyclodextrine and alpha-amylase thermostability; Morand P et al.; The cleavage of beta-cyclodextrine by sodium periodate at the seven 2-3 diols of the glucose unit gives rise to the polyaldehyde 1, used to modify alpha-amylase . The reductive modification of alpha-amylase from Bacillus licheniformis reduced the number of reactive lysine groups from 8 to 3.5 per mol of enzyme with an activity loss of 25% and increased the half-life at 80 degrees C from 4.7 to 7.0 minutes. J Biol Chem, 1991 Sep 5, 266(25), 16356 - 62 Molecular dissection of translation initiation factor IF2 . Evidence for two structural and functional domains; Gualerzi CO et al.; By means of limited proteolysis of Bacillus stearothermophilus initiation factor IF2 and genetic manipulation of its structural gene, infB, we have been able to produce (or hyperproduce) and purify two polypeptide fragments corresponding to two structurally and functionally separate domains of the protein . The first is the G-domain (approximately 41 kDa), which makes up the central part of the molecule and contains the conserved structural elements found in all GTP/GDP-binding sites of G-proteins . This domain is resistant to proteolysis in the presence of GTP or GDP, retains the capacity to interact with the 50 S subunit, binds weakly to the 30 S subunit, and displays ribosome-dependent GTPase activity with an approximately 2-fold higher Km for GTP and the same Vmax as compared with intact IF2 . The second is the C-domain (approximately 24 kDa), which corresponds to the COOH-terminal part of IF2 and constitutes an extraordinarily compact domain containing the fMet-tRNA binding site of IF2 . In spite of its negligible affinity for the ribosomes, the C-domain weakly stimulates the ribosomal binding of fMet-tRNA, presumably by affecting the conformation of the initiator tRNA molecule. J Appl Bacteriol, 1991 Sep, 71(3), 285 - 8 Effect of simultaneous application of heat and pressure on the survival of bacterial spores; Mallidis CG et al.; The effect of simultaneous application of moderate hydrostatic pressure (10-300 atm) and heat on the survival of the Bacillus stearothermophilus spores in a flow-through system was investigated . A high heterogeneity of the sensitization of spores to heat by pressure was found . A higher degree of reduction of heat resistance was observed at the low than at the high temperatures tested . The simultaneous application of moderate pressure and heat can not be applied for the preservation of liquid foods due to the extreme heterogeneity of spore sensitization to heat by pressure. J Appl Bacteriol, 1991 Sep, 71(3), 202 - 6 Formulation of a flowable liquid concentrate of Bacillus thuringiensis serotype H-14 spores and crystals as mosquito larvicide; Ejiofor AO et al.; Bacillus thuringiensis serotype H-14 spores and crystals, produced in 51 fermenters, were centrifuged and resuspended in emulsified palm olein to give 3.2 x 10(11) colony forming units (cfu)/ml . The suspension was mixed with a cassava-molasses-palm olein-charcoal (CMPC-2) mixture which served as the carrier, adhesive, dispersant and protectant . The final concentration of the formulation was 3.2 x 10(9) cfu/ml . The lethal concentrations capable of killing 50% of the test population (LC50) of CMPC-2 during 0, 1 and 2 years of storage at 32 +/- 4 degrees C were 0.056, 0.058 and 0.058 mg/ml respectively as against 0.054, 0.051 and 0.054 mg/ml for the Institut Pasteur Standard-1978 (IPS-78) during the corresponding period . The chi 2 tests showed that the results were homogeneous at P = 0.05 . The relative potencies of the preparations were 964.3, 879.3 and 931 International toxic units (ITU) Aedes aegypti as compared with the 1000 ITU assigned to IPS-78 . At 95% confidence limits there was no significant difference between the potencies of CMPC-2 and IPS-78 . Field tests showed that CMPC-2 provided between 87.5 and 100% control of natural populations of Aedes spp . and Cutex spp . Sedimentation tests showed that CMPC-2 settled markedly during storage . This, therefore, required that the product be thoroughly shaken before use. Antimicrob Agents Chemother, 1991 Sep, 35(9), 1933 - 6 In vitro activity of the new difluorinated quinolone sparfloxacin (AT-4140) against Mycobacterium tuberculosis compared with activities of ofloxacin and ciprofloxacin; Rastogi N et al.; MICs of the new fluoroquinolone drugs ofloxacin, ciprofloxacin, and sparfloxacin (AT-4140) for 10 strains of Mycobacterium tuberculosis were determined by using both a BACTEC radiometric method and testing on solid 7H11 agar medium . Radiometric MICs by 7H12 broth testing ranged from 0.5 to 1.0, 0.25 to 0.5, and 0.1 to 0.2 microgram/ml for ofloxacin, ciprofloxacin, and sparfloxacin respectively, whereas MICs in solid medium ranged from 0.5 to 1.0, 0.5 to 1.0, and 0.2 to 0.5 microgram/ml, respectively . The bactericidal action of the quinolones compared with their reported peak concentrations in human serum showed that sparfloxacin is the most bactericidal, followed by ciprofloxacin and ofloxacin . Our results suggest the potential of the new difluorinated quinolone sparfloxacin for use against the tubercle bacillus and indicate that further determination of its antimycobacterial spectrum and intracellular efficacy may be fruitful. Quintessence Int, 1991 Sep, 22(9), 721 - 6 Evaluation of a dental unit with a built-in decontamination system; Douglas CW et al.; The efficacy of a dental unit equipped with a system that disinfects and sterilizes the water tubing by flushing with glutaraldehyde was evaluated by inserting Bacillus megaterium spores and Pseudomonas and Moraxella species into the water tubing . Up to 10(8) Pseudomonas and Moraxella organisms were killed during the disinfection cycle, but Bacillus megaterium spores were not . Up to 10(5) spores were eradicated by the sterilization cycle, although the system did not consistently kill 10(8) spores . The water tubing of the new unit was not naturally colonized by water bacteria during an 8-month period prior to the study . Evidence suggested that this was due to antimicrobial activity associated with the plastic tubing; therefore, microbial contamination of new dental units, irrespective of their design, would not be expected, until the inhibitory factor in the plastic tubing has leached out. Kekkaku, 1991 Sep, 66(9), 577 - 87 {Preventable tuberculosis cases in Japan--a new approach to the assessment of tuberculosis control problems}; Ahiko T; I defined cases of tuberculosis which could not be prevented from infection or development of disease among infected, or could not be detected in the early stages as "preventable cases" in order to evaluate tuberculosis control efforts in the community, Japan . Among 241 bacteriologically confirmed cases with pulmonary tuberculosis newly registered from 1988 through 1989 in Yamagata Prefecture, 80 (33%) were defined as preventable cases by observing their course and the process of diagnosis . That is to say, one-third of bacillary cases could have been prevented in Yamagata where the incidence of tuberculosis was lowest in Japan, if existing prevention and control methods had been effectively used . Causes of prevention failure were investigated in detail . The most common cause was tardy detection of cases, especially due to delayed confirmation of diagnosis (so called "doctor's delay") . The delayed confirmation of diagnosis resulted from neglecting chest X-ray and sputum examination and from ignoring high risk groups . In the younger age group, it was mainly attributed to insufficient family contact examinations . Tuberculin skin tests are necessary not only for those aged 15 years and younger but also for those aged 16-29, when they are found to be household contacts of smear-positive cases . An evaluation of tuberculosis control program defining preventable cases would be a beneficial approach to the surveillance of tuberculosis. Semin Dermatol, 1991 Sep, 10(3), 194 - 8 Bacillary angiomatosis: a systemic opportunistic infection with prominent cutaneous manifestations; LeBoit PE; Bacillary angiomatosis is an opportunistic infection with systemic manifestations . Although most cases have occurred in human immunodeficiency virus-positive patients, other immunosuppressed patients, and even seemingly immunocompetent individuals, can become infected . In addition to the well-characterized cutaneous manifestations, visceral involvement can occur and may be the only locus of infection . Lymphadenopathy, bone or soft-tissue masses, fever, and hepatosplenomegaly can be presenting signs . The causative bacterium is still unidentified, but resemblances to the rickettsiae, Rochilamea quintana, the recently identified cat-scratch disease bacillus, and Bartonella bacilliformis have been noted by various investigators . Systemic disease is treatable and can be cured with antibiotic therapy. Infect Immun, 1991 Sep, 59(9), 2864 - 9 Effects of activated macrophages on Mycobacterium leprae; Ramasesh N et al.; Five alternative methods were used to explore in vitro the effects of normal and activated murine macrophages on the metabolic well-being of intracellular Mycobacterium leprae: fluorescein diacetate-ethidium bromide staining, ATP content, synthesis of phenolic glycolipid 1, and two techniques to quantitate oxidation of palmitic acid . In relatively short-term experiments (7 to 10 days), each of these procedures provided strong evidence that activated macrophages exerted a deleterious effect on the leprosy bacillus . These findings appear to confirm the contention that activated macrophages underlie host resistance to clinical leprosy and limitation of M . leprae growth in paucibacillary leprosy. Eur J Biochem, 1991 Sep 1, 200(2), 545 - 51 Competitive inhibition of liver glucokinase by its regulatory protein; Vandercammen A et al.; The regulatory protein of rat liver glucokinase (hexokinase IV or D) behaved as a fully competitive inhibitor of this enzyme when glucose was the variable substrate, i.e . it increased the half-saturating concentration of glucose as a linear function of its concentration without affecting V (velocity at infinite concentration of substrate) . The inhibition by the regulatory protein and that by palmitoyl-CoA were synergistic with that by N-acetyl-glucosamine, indicating that the two former inhibitors bind to a site distinct from the catalytic site . In contrast, the effects of the regulatory protein and palmitoyl-CoA were competitive with each other, indicating that these two inhibitors bind to the same site . The regulatory protein exerted a non-competitive inhibition with respect to Mg-ATP at concentrations of this nucleotide less than 0.5 mM . At higher concentrations, the latter antagonized the inhibition by the regulatory protein partly by decreasing the apparent affinity for fructose 6-phosphate . The following anions inhibited glucokinase non-competitively with respect to glucose: Pi, sulfate, I-, Br-, No3-, Cl-, F- and acetate . Pi and sulfate, at concentrations in the millimolar range, decreased the inhibition by the regulatory protein by competing with fructose 6-phosphate . Monovalent anions also antagonized the inhibition by the regulatory protein with the following order of potency: I- greater than Br- greater than NO3- greater than Cl- greater than F- greater than acetate and their effect was non-competitive with respect to fructose 6-phosphate . Glucokinase from Buffo marinus and pig liver were, like the rat liver enzyme, inhibited by the regulatory protein, as well as by palmitoyl-CoA at micromolar concentrations . In contrast, neither compound inhibited hexokinases from rat brain, beef heart or yeast, or the low-Km specific glucokinase from Bacillus stearothermophilus. Urology, 1991 Sep, 38(3), 271 - 9 Multicenter study of superficial bladder cancer treated with intravesical bacillus Calmette-Guerin or adriamycin . Results of long-term follow-up; Khanna OP et al.; We evaluated 158 cases of patients with superficial bladder cancers (Stages Ta, T1, and Tis) . These cases were treated with either intravesical bacillus Calmette-Guerin (BCG) (Tice strain) or Adriamycin (ADR), in a multicenter, nonrandomized study . One hundred thirty-one of these patients were followed up; the results continue to show a higher percentage of initial complete remissions with BCG (68%) than with ADR (57%) . With additional therapy, both BCG and ADR achieved complete remission in 83 percent of the patients . When 7 failures with patients taking ADR were switched to BCG and the disease cleared, the rate of complete remission for BCG rose to 85 percent . The recurrence rate per 100 patient-months was only slightly different for BCG (0.9) and ADR (0.8) . The percentage of progressions continued to be higher for BCG (8%) than for ADR (5%) . Cystectomies were performed in 2.5 percent of the BCG patients . Using the Cox regression model with covariates, we found drug treatment, tumor grade, and sex to be statistically significant in determining failures throughout the protocol . Although both BCG and ADR were effective over the course of the study, BCG is the drug of choice for residual tumor (Stages T1 and Tis). J Urol, 1991 Sep, 146(3), 700 - 2; discussion 702-3 The management of transitional cell carcinoma in solitary renal units; Schoenberg MP et al.; Ten patients with urothelial malignancies involving a solitary functioning renal unit were treated at our center for an average of 24 months or until death . These patients were all managed by parenchyma-sparing methods, including percutaneous as well as ureteroscopic tumor resection . Of our patients 9 have received adjunctive chemotherapy in the form of bacillus Calmette-Guerin instillations . At the time of this report 5 of our patients were alive without evidence of disease, 4 were alive with evidence of either residual or recurrent neoplasia and 1 was dead of disease 5 years after original presentation . Patients with higher grade tumors or carcinoma in situ did less well than patients with low grade disease . We present an analysis of our experience with this complex patient population and discuss the implications of these data within the context of a growing literature on the topic of upper tract urothelial malignancy. Bioorg Khim, 1991 Sep, 17(9), 1188 - 92 {BsoAI--a new site specific endonuclease from Bacillus coagulans}; Sokolov NN et al.; A new site-specific endonuclease was detected in toluene lysates of Bacillus coagulans AUCM B-732 and designated as BcoAI . The enzyme was purified by fractionation of the cell-free extract in the two-phase PEG/dextran system followed by chromatography on DEAE-sepharose and phosphocellulose and shown to be free of nonspecific nucleases and phosphatases . BcoAI has three cleavage sites on lambda DNA, but does not cleave SV40, pBR322 and pUC19 DNA . BcoAI recognizes the sequence 5' CAC decreases GTG 3' on double-stranded DNA and cleaves it as indicated by the arrow to yield blunt-ended DNA fragments . Thus, BcoAI is a true isoschizomer of PmaCI from Pseudomonas maltophila C. Indian J Malariol, 1991 Sep, 28(3), 167 - 70 Acute toxicity of certain organochlorine, organophosphorus, synthetic pyrethroid and microbial insecticides to the mosquito fish Gambusia affinis (Baird and Girard); Mittal PK et al.; Acute toxicity of certain organochlorine, organophosphorus, synthetic pyrethroid and microbial insecticides to the mosquito fish Gambusia affinis were determined to collect baseline data for selecting the resistant strains of the fish . The synthetic pyrethroid, Lambdacyhalothrin was most toxic to the fish (LC50 = 0.0022 ppm), followed by deltamethrin, cypermethrin and fenvalerate . Organochlorine insecticides, DDT and gamma-HCH, were less toxic than the pyrethroids, and these were followed by organophosphorus insecticides, malathion, fenthion, monocrotophos and temephos . The last two insecticides were least toxic among the different chemical insecticides (LC50 greater than 80 ppm ai) . The microbial insecticide ABG-6262 (Vectolex 2.5 AS), a Bacillus sphaericus preparation, was totally harmless to the fish at 2500 microliters/l up to one week. Indian J Malariol, 1991 Sep, 28(3), 147 - 50 Isolation and laboratory evaluation of an indigenous strain of Bacillus sphaericus (9001); Gupta DK et al.; An indigenous strain of Bacillus sphaericus H5a (9001), possessing high insecticidal properties, was isolated from diseased larvae of Culex species . This strain in comparison with the known strains of B . sphaericus, i.e., 1593 and 2362, was found to be promising against the fourth instar larvae of Anopheles culicifacies, An . stephensi, An . subpictus, Aedes aegypti and Culex quinquefasciatus . B . sphaericus 9001 is highly stable and virulent through 25 successive transfers and thus can be effectively used as a biocontrol agent against immature stages of mosquitoes. Southeast Asian J Trop Med Public Health, 1991 Sep, 22(3), 429 - 35 Toxicity of Bacillus sphaericus strain 2362 on Mansonia spp . larvae; Petcharat J; The efficiency of Bacillus sphaericus strain 2362 (Vectolex) as larvicide against Mansonia spp . was studied . Bioassay studies showed that the toxicity of B . sphaericus on both age groups (I-II instar and III-IV instar) of Mansonia spp . larvae occurred within 24 hours . Probit analysis revealed that LC100 (one hundred per cent lethal concentration) for both age groups of M . boneae were higher than those of M . dives . Small scale field trials were done at Kreng Village, Cha-uat District, Nakhon Si Thammarat Province, one of the most serious filarial infected areas . It was indicated that 100% kill of Mansonia spp . larvae in the field occurred within 9 days after the larvicide application . When a dose of 5 times of LC100 value was used, 100% control was achieved up to about one month. Southeast Asian J Trop Med Public Health, 1991 Sep, 22(3), 426 - 8 Laboratory evaluation of Bacillus thuringiensis H-14 against Aedes aegypti larvae in the northeast region of Thailand; Pipitgool V et al.; Laboratory bioassays using a preparation of Bacillus thuringiensis H-14 (Bt.H-14), namely Skeetal were conducted to determine their effectiveness against late 3rd/early 4th instar larvae of Aedes aegypti . The larvae were collected from municipal areas in 7 provinces, namely Burirum, Roi-Et, Khon Kaen, Ubol Ratchatani, Nakorn Phanom, Surin and Nakorn Ratchasima, in the Northeast of Thailand . It was found that for Skeetal, LC50 ranged from 128 to 151 nl/l (average 143) and LC90 ranged from 254 to 289 nl/l (average 275) . The mortality rate of Ae . aegypti larvae in the 7 provinces did not differ significantly (p greater than 0.05) at a concentration of 300 nl/l . The result of the bioassays show that the preparation of Bt.H-14 is very effective against Ae . aegypti larvae in Northeast of Thailand and the mosquito larvae in the various areas were nearly equal in susceptibility to Bt.H-14. Yale J Biol Med, 1991 Sep-Oct, 64(5), 481 - 98 Consumption, silicosis, and the social construction of industrial disease; Rosner D et al.; In the wake of the bacterial revolution after Robert Koch identified the tuberculosis bacillus, medical and public health professionals classified the various forms of consumption and phthisis as a single disease--tuberculosis . In large measure, historians have adopted that perspective . While there is undoubtedly a great deal of truth in this conceptualization, we argue that it obscures almost as much as it illuminates . By collapsing the nineteenth-century terms phthisis and consumption into tuberculosis, we maintain that historians have not understood the effect of non-bacterial consumption on working-class populations who suffered from the symptoms of coughing, wasting away, and losing weight . In this essay, we explore how, in the nineteenth century, what we now recognize as silicosis was referred to as miners' "con," stonecutters' phthisis, and other industry-specific forms of phthisis and consumption . We examine how the later and narrower view of the bacterial origins of tuberculosis limited the medical professions' ability to diagnose and understand diseases caused by industrial dust . This paper explores the contention that developed at the turn of the century over occupational lung disease and tuberculosis and the circumstances that led to the unmasking of silicosis as a disease category. Zhonghua Nei Ke Za Zhi, 1991 Sep, 30(9), 572 - 4, 596 {Clinical effect and laboratory observation of ofloxacin in the treatment of typhoid fever, bacillary dysentery and gonorrhea}; Zou QY et al.; Ofloxacin was used in the treatment of 20 cases of typhoid fever, 32 cases of bacillary dysentery and 50 cases of gonococcal infection . Altogether 102 cases were treated, 53 being male and 49 female . The daily dosage was 400 mg to 600 mg, divided into two times . The result showed that the clinical effective rate for typhoid fever, bacillary dysentery and gonococcal infections was 100%, 97% and 94% respectively, while the bacterial eradication rate was 100%, 100% and 94% respectively . the bacterial eradication rate was 100%, 97% and 94% respectively . The side effects were mild in degree . The authors are of the opinion that since ofloxacin can be administered orally with only two times a day, its absorption is nearly complete and the cure rate is high, it should be considered as the drug of choice in the treatment of typhoid fever, bacillary dysentery and gonorrhea, especially in the drug resistant cases . It is suggested that this drug be used more widely. Chem Pharm Bull (Tokyo), 1991 Sep, 39(9), 2468 - 70 Isolation of a new antitumor substance from Bacillus stearothermophilus; Kohama Y et al.; A new antitumor substance, BS-1, was isolated from the autolysate and culture filtrate of Bacillus stearothermophilus UK563 by ethylacetate extraction and HPLC . BS-1 inhibited the proliferation of mouse macrophage-like cells, P388-D1 (IC50: 4 micrograms/ml) and mouse mastocytoma, P-815 (IC50: 0.6 microgram/ml), but not that of Balb/c 3T3. J Am Mosq Control Assoc, 1991 Sep, 7(3), 420 - 3 Delayed mortality and morphogenetic anomalies induced by the microbial control agent Bacillus thuringiensis ser . (H-14) in Culex quinquefasciatus; Mulla MS et al.; Sublethal concentrations of Bacillus thuringiensis ser . H-14 were applied to early 4th-instar larvae of Culex quinquefasciatus to assess mortality and morphogenetic aberrations in larvae, pupae and adults . At the 24 h LC10, LC25, LC50 and LC80, additional mortality occurred in surviving larvae beyond a 24 h exposure period . The cumulative mortalities increased daily and the overall mortality of larvae up to 7 days posttreatment were 12, 73, 82 and 96% at the indicated concentrations, respectively . Delayed mortality also occurred in the pupal and adult stages . Morphogenetic aberrations were noted in dead larvae and pupae but were rare in the adults . These aberrations are categorized and described . There was little or no delayed effect on the survivorship or fecundity of adults, but in all the treatments the number of emerging males was higher than females . The sex ratio in check adults was 1:1. J Am Mosq Control Assoc, 1991 Sep, 7(3), 412 - 9 Delayed mortality and morphogenetic anomalies induced in Culex quinquefasciatus by the microbial control agent Bacillus sphaericus; Mulla MS et al.; Two preparations of Bacillus sphaericus 2362 were studied for their biological activity, delayed mortality and the induction of morphogenetic aberrations in larvae, pupae and adults of Culex quinquefasciatus . Longevity and fecundity of adult mosquitoes were also assessed . A dosage response line for B . sphaericus was established against 4th-instar larvae and sublethal concentrations (48 h LC50 and lower) were used against these larvae . Sublethal concentrations of B . sphaericus induced delayed mortality in larvae, pupae and adults . The magnitude of mortality increased in succeeding cohorts and developmental stages resulting from the surviving larvae . Only 10 and 25% overall emergence of viable adults occurred in the sublethal treatments (LC25) of 2 B . sphaericus preparations . The range of successful adult emergence was over 94% in the controls . A wide range of external morphogenetic aberrations in dead larvae, pupae and adults were noted . These aberrations and gross morphological features were quite similar to those reported for certain insect growth regulators . Sublethal concentrations had no marked effect on longevity of adults, egg deposition and hatch. Indian J Med Res, 1991 Sep, 93, 318 - 23 Production & formulation of Bacillus thuringiensis var . israelensis & B . sphaericus 1593; Desai SY et al.; Three fermentation media each for bulk growth of B . thuringiensis var . israelensis and B . sphaericus 1593 were formulated using defatted groundnut cake (Arachis hypogea) as the first nitrogen source and gram flour (Cicer arientinum), soy bean (Glycine max) and defatted milk powder as the second nitrogen source . Medium containing gram flour showed highest toxicity (14.45 micrograms/l) in case of B . thuringiensis var . israelensis whereas medium containing milk powder was found to be highly toxic with B . sphaericus 1593 (51.39 micrograms/l) . Sustained release floating pellet formulations of B . thuringiensis var . israelensis and B . sphaericus 1593 exhibited toxicity of 77 per cent and above for 42 days at a dose of 500 micrograms/l for 4th instar larvae of Culex pipiens quinquefasciatus Say. J Clin Microbiol, 1991 Sep, 29(9), 1777 - 9 Pneumonia and empyema infection associated with a Bacillus species that resembles B . alvei; Coudron PE et al.; An organism resembling Bacillus alvei was isolated from the lung and pleural fluid of an immunocompetent patient . The isolate differed from the type strain of B . alvei in its ability to reduce nitrate and its inability to produce dihydroxyacetone and acetylmethylcarbinol . The isolate was resistant to ciprofloxacin and showed intermediate susceptibility to vancomycin. Appl Environ Microbiol, 1991 Sep, 57(9), 2767 - 70 Enhancement of soybean nodulation by Bacillus cereus UW85 in the field and in a growth chamber; Halverson LJ et al.; Seed treatments with Bacillus cereus UW85 increased nodulation of soybeans in three field seasons and in three different sterilized soils in the growth chamber . In the field, 28 and 35 days after planting, UW85-treated plants had 31 to 133% more nodules than untreated plants . From 49 days after planting until seed harvest, there were no significant differences between nodulation of UW85-treated plants and untreated control plants . In the growth chamber, in sterilized soil-vermiculite mixtures, at 28 days after planting, UW85 seed treatments enhanced nodulation by 34 to 61%, indicating that the increase in nodulation was not dependent on the soil flora. Appl Environ Microbiol, 1991 Sep, 57(9), 2656 - 65 Utility of phenomenological models for describing temperature dependence of bacterial growth; Heitzer A et al.; We compared three unstructured mathematical models, the master reaction, the square root, and the damage/repair models, for describing the relationship between temperature and the specific growth rates of bacteria . The models were evaluated on the basis of several criteria: applicability, ease of use, simple interpretation of model parameters, problem-free determination of model parameters, statistical evaluation of goodness of fit (chi 2 test), and biological relevance . Best-fit parameters for the master reaction model could be obtained by using two consecutive nonlinear least-square fits . The damage/repair model proved to be unsuited for the data sets considered and was judged markedly overparameterized . The square root model allowed nonproblematical parameter estimation by a nonlinear least-square procedure and, together with the master reaction model, was able to describe the temperature dependence of the specific growth rates of Klebsiella pneumoniae NCIB 418, Escherichia coli NC3, Bacillus sp . strain NCIB 12522, and the thermotolerant coccobacillus strain NA17 . The square root and master reaction models were judged to be equally valid and superior to the damage/repair model, even though the square root model is devoid of a conceptual basis. Scand J Immunol, 1991 Sep, 34(3), 365 - 72 MPB 64 possesses 'tuberculosis-complex'-specific B- and T-cell epitopes; Andersen AB et al.; We have developed monoclonal antibodies (MoAb) reactive with a protein from Mycobacterium tuberculosis of apparent molecular mass 24 kDa . This protein was shown to be identical with MPB 64 (Harboe et al.,) MoAb bound to four different epitopes of which two were restricted to the 'tuberculosis complex' and two were also found in mycobacteria not belonging to the 'tuberculosis complex' . The cross-reactive MoAb demonstrate that MPB 64 is present in more mycobacterial species than previously assumed . MPB 64 was shown to induce strong delayed type hypersensitivity (Dth) reactions in outbred guinea pigs immunized with M . tuberculosis and M . bovis bacille Calmette-Guerin (BCG) . No reaction was observed in animals immunized with mycobacteria not belonging to the 'tuberculosis complex' . The Dth-inducing capacity of MPB64 was compared with that of another 24 kDa protein purified from M . tuberculosis and of the previously described 38 kDa protein . The Dth responses to these three antigens were further analysed in four inbred guinea pig strains . A genetic restriction of the ability of the animals to respond to MPB 64 as well as to the 38 kDa protein was observed. Microbiol Rev, 1991 Sep, 55(3), 425 - 36 Bacillus sphaericus as a mosquito pathogen: properties of the organism and its toxins; Baumann P et al.; In the course of sporulation, Bacillus sphaericus produces an inclusion body which is toxic to a variety of mosquito larvae . In this review we discuss the general biology of this species and concentrate on the genetics and physiology of toxin production and its processing in the midgut of the larval host . The larvicide of B . sphaericus is unique in that it consists of two proteins of 51 and 42 kDa, both of which are required for toxicity to mosquito larvae . There is a low level of sequence similarity between these two proteins, which differ in their sequences from all the other known insecticidal proteins of Bacillus thuringiensis . Within the midgut the 51- and 42-kDa proteins are processed to proteins of 43 and 39 kDa, respectively . The conversion of the 42-kDa protein to a 39-kDa protein results in a major increase in toxicity; the significance of the processing of the 51-kDa protein is not known . In contrast to the results with mosquito larvae, the 39-kDa protein is alone toxic for mosquito-derived tissue culture-grown cells, and this toxicity is not affected by the 51-kDa protein or its derivative, the 43-kDa protein . Comparisons of larvae from species which differ in their susceptibility to the B . sphaericus toxin indicate that the probable difference resides in the nature of the target sites of the epithelial midgut cells and not in uptake or processing of the toxin . A similar conclusion is derived from experiments involving tissue culture-grown cells from mosquito species which differ in their susceptibility to the B . sphaericus toxin. Biochimie, 1991 Sep, 73(9), 1179 - 86 Glycosyl-phosphatidylinositol anchored acetylcholinesterase as substrate for phosphatidylinositol-specific phospholipase C from Bacillus cereus; Stieger S et al.; We investigated the enzymatic properties of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus towards glycosyl-phosphatidylinositol anchored acetylcholinesterase (AChE) from bovine erythrocytes and Torpedo electric organ as substrate . The conversion of membrane from AChE to soluble AChE by PI-PLC depended on the presence of a detergent and of phosphatidylcholine . In presence of mixed micelles containing Triton X-100 (0.05%) and phosphatidylcholine (0.5 mg/ml) the rate of AChE conversion was about 3 times higher than in presence of Triton X-100 alone . Furthermore, inhibition of PI-PLC occurring at Triton X-100 concentrations higher than 0.01% could be prevented by addition of phosphatidylcholine . Ca2+, Mg2+ and sodium chloride had no effect on PI-PLC activity in presence of phosphatidylcholine and Triton X-100, whereas in presence of Triton X-100 alone sodium chloride largely increased the rate of AChE conversion . Determination of kinetic parameters with three different substrates gave Km-values of 7 microM, 17 microM and 2 mM and Vmax-values of 0.095 microM.min-1, 0.325 microM.min-1 and 56 microM.min-1 for Torpedo AChE, bovine erythrocyte AChE and phosphatidylinositol, respectively . The low Km-values for both forms of AChE indicated that PI-PLC not only recognized the phosphatidylinositol moiety of the anchor but also other components thereof. Biochem J, 1991 Sep 1, 278 ( Pt 2), 461 - 3 A study of human erythrocyte acetylcholinesterase inhibition by chlorpromazine; Spinedi A et al.; Membrane-bound acetylcholinesterase (AChE) from the human erythrocyte is inhibited by chlorpromazine (CPZ) in a concentration range within this amphiphilic drug has been demonstrated to interact with erythrocyte membranes, causing a large spectrum of physical and structural effects; membrane solubilization with 0.5% Triton X-100 results in a complete loss of CPZ inhibitory potency . Although these observations might suggest a role of membrane lipid environment in mediating human erythrocyte AChE inhibition, we observed that CPZ retains its full inhibitory effect on the fraction of enzyme (5-6% of total) that is solubilized from erythrocytes upon treatment with phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus thuringiensis; furthermore, Triton X-100 is able to reverse the CPZ effect also in the case of PI-PLC-solubilized enzyme . These results demonstrate unequivocally that CPZ inhibits human erythrocyte AChE through direct molecular interaction . The inhibition kinetics displayed by CPZ on human erythrocyte AChE are dependent on drug concentration: evidence is provided that this phenomenon may be related to formation of CPZ micellar aggregates. Lett Appl Microbiol, 1991 Sep, 13(3), 171 - 4 The linear PCR reaction: a simple and robust method for sequencing amplified rRNA genes; Embley TM; The linear polymerase chain reaction was used to sequence amplified RNA genes from strains of Bacillus, Thermus and Legionella . The technique described is simple and reproducible and it works well with double standard product which has been PEG precipitated directly from PCR reactions. Agric Biol Chem, 1991 Sep, 55(9), 2251 - 8 Purification and properties of a novel surface-active agent- and alkaline-resistant protease from Bacillus sp . Y; Shimogaki H et al.; In the course of a search for an alkaline stable protease for industrial use, an alkaline protease (protease BYA) was isolated from an alkalophilic Bacillus sp . Y, and its properties were characterized . Its optimum pH was pH 10.0-12.5, when casein was used as a substrate . In addition to the stability of protease BYA at pH 6.5-13.0, it was also very stable towards various surface-active agents, such as sodium dodecyl sulfate and sodium linear alkylbenzene sulfonate . Protease BYA was most active at 70 degrees C . The isoelectric point (pI) of protease BYA was about 10.1 . Protease BYA was characterized as a serine protease because of its sensitivity to phenylmethanesulfonyl fluoride and diisopropyl fluorophosphate . The protease seems to be related to proteases of the subtilisin family, such as subtilisin BPN', subtilisin Carlsberg, and No . 221 protease. Arch Biochem Biophys, 1991 Aug 15, 289(1), 167 - 79 Physical studies on membrane lipids of Bacillus stearothermophilus temperature and calcium effects; Jurado AS et al.; Bacillus stearothermophilus was grown at the optimal temperature range (center, 65 degrees C), below it (48 and 55 degrees C), and above it (68 degrees C), in a complex medium with or without 2.5 mM Ca2+ . The Ca(2+)-supplement improves growth at sub- and supraoptimal temperatures and extends it to higher temperatures (Jurado et al . (1987) J . Gen . Microbiol . 133, 507-513) . The phospholipid composition of cultures obtained in the different growth conditions was studied . Phosphatidylethanolamine was always the major phospholipid (40 to 50% of the total phospholipid) . Diphosphatidylglycerol, phosphatidylglycerol, a phosphoglycolipid (pgl) and two minor phospholipids (not identified) were also found in the polar lipid extract . The pgl shows a threefold concentration increase as the growth temperature raises from 48 to 68 degrees C . The thermotropic behavior of membrane lipids was studied by differential scanning calorimetry (DSC) and by means of two fluorescent probes of fluidity, 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1,3-di(2-pyrenyl)propane (2Py(3)2Py) . The results reveal similar features and clearly show a shift of the temperature range of the phase transition to higher values and an increased structural order of the bilayer, as the growth temperature rises from 55 to 68 degrees C, but an opposite effect was observed from 48 to 55 degrees C . Although the Ca(2+)-supplement to the growth medium has no detectable effect, the addition of Ca2+ to the buffer of liposomes (Ca(2+)-liposomes) has a significant ordering effect at all growth temperatures . These liposomes show a shift of the transition range to higher temperatures and the fluorescent parameters (DPH polarization and intramolecular excimerization of the 2Py(3)2Py) detected an order increase of the probes environment, along and above the main phase transition . Spectra of 31P-NMR and polarized light microscopy clearly show that the lipid extracts exhibit, in all the conditions, typical lamellar phase geometry . We concluded that B . stearothermophilus controls the membrane lipid composition to compensate for the destabilizing effect of high temperatures on the membrane organization or to provide an appropriate packing of phospholipid molecules in a stable bilayer . At high temperatures, Ca(2+)-stimulatory effect on growth is presumably due to a direct Ca2+ interaction with the membrane phospholipids, inducing an increased structural order on the bilayer . The increase of the phase transition temperature in the total lipid extracts as compared with the respective polar lipid fractions probably indicates a stabilizing effect of neutral lipids on membrane bilayers. Carbohydr Res, 1991 Aug 12, 215(1), 127 - 36 Characterization of five isomers of branched cyclomaltoheptaose (beta CD) having degree of polymerization (d.p.) = 9: Reinvestigation of three positional isomers of diglucosyl-beta CD; Koizumi K et al.; It has been confirmed by methylation analyses and chemical syntheses that three isomers of branched cyclomaltoheptaose (beta CD) isolated from the mother liquors of a large-scale preparation of beta CD with Bacillus ohbensis cyclomaltodextrin glucanotransferase are 6(1),6(4)-di-O-(alpha-D-glucopyranosyl)-cyclomaltoheptaose (1), 6(1),6(3)-di-O-(alpha-D-glucopyranosyl)-cyclomaltoheptaose (2), and 6-O-(alpha-isomaltosyl)-cyclomaltoheptaose (4) instead of 6(1),6(2)-di-O-(alpha-D-glucopyranosyl)-cyclomaltoheptaose (3), which was erroneously characterized in an earlier paper . Compound 3 has been newly isolated from a glucosyl-beta CD mixture prepared by hydrolysis with glucoamylase of a maltosyl-beta CD mixture, synthesized from maltose and beta CD through the reverse action of pullulanase . Chromatographic behavior and spectral data (13C-n.m.r . and f.a.b.-m.s.) of these isomers of branched beta CD (1-4), as well as those of another isomer prepared by the reverse action of hydrolytic enzymes, 6-O-(alpha-maltosyl)-cyclomaltoheptaose (5), were compared. J Biol Chem, 1991 Aug 5, 266(22), 14310 - 6 Stopped-flow and rapid-scan studies of the redox behavior of cytochrome aco from facultative alkalophilic Bacillus; Orii Y et al.; Cytochrome aco purified from an alkalophilic bacterium grown at pH 10 contains hemes a, b, and c as prosthetic groups, and their redox behavior was examined by using stopped-flow and rapid-scan techniques . Under anaerobic conditions the reduction of both heme a and c moieties with dithionite proceeded exponentially but with different rates, usually the former being reduced about 4 times faster than the latter . The reduction of protoheme was much slower, and a time-difference spectrum for this species was of a high spin type with absorption peaks at 433, 557, and 609 nm . Only the protoheme combined with CO, fulfilling the criteria for cytochrome o . Potentiometric titrations determined a midpoint potential of c heme to be 95 mV at pH 7.0 and 25 degrees C and suggested the presence of two forms of a heme with midpoint potentials of 250 and 323 mV . Cytochrome aco utilizes ascorbate plus N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) to reduce oxygen relatively rapidly without added cytochrome c (Qureshi, M . H., Yumoto, I., Fujiwara, T., Fukumori, Y., Yamanaka, T . (1990) J . Biochem . 107, 480-485) . During the steady state, however, heme a stayed almost fully reduced in contrast to a partial reduction of heme c . Even after exhaustion of the dissolved oxygen the extent of reduction of heme c was 60-70% that attained by the dithionite reduction . When ascorbate plus TMPD-reduced cytochrome aco was exposed to oxygen the reduced heme c was oxidized rapidly whereas the oxidation of reduced a heme was negligibly slow . The full reduction of heme a during the steady state and its extremely slow oxidation rendered participation of heme a in the oxidase reaction less likely . A novel peak appearing transiently around 567 nm during the reaction was tentatively ascribed to an intermediate form of protoheme, or o heme, which was thus supposed to react directly with molecular oxygen . These results suggest strongly that the main electron transfer pathway would be c----o----oxygen . A possible role of a in regulating the electron flow through the main pathway and its functional relationship to a heme in the aa3-type cytochrome oxidase were discussed. Semin Vet Med Surg (Small Anim), 1991 Aug, 6(3), 199 - 202 Cat scratch disease: no longer a diagnostic dilemma; Margileth AM; Cat scratch disease is a relatively common cause of localized lymphadenopathy with about 80% of cases occurring in children . This self-limited infection is caused by a small pleomorphic gram-negative bacillus that has been identified in ocular granulomas, skin and lymph node specimens . Unusual manifestations of the disease, such as the oculoglandular disease of Parinaud, encephalopathy, or severe systemic disease, occur in about 12% of patients . Management consists of symptomatic treatment, occasionally aspiration of a node or selected antibiotic therapy in moderate to severely ill patients . Persistence of adenopathy for several months in a generally healthy patient with gradual, spontaneous resolution of the enlarged node is the natural course. J Appl Bacteriol, 1991 Aug, 71(2), 130 - 3 Bacteria in food packaging paper and board; Vaisanen OM et al.; The bacteria of food packaging paper and board were studied . Most of the aerobic strains were spore-formers; members of the genus Bacillus with B . cereus group (B . cereus, B . mycoides, B . thuringiensis), B . polymyxa group (B . polymyxa, B . circulans, B . macerans, B . pabuli), B . brevis and B . licheniformis predominated . The main source of spore-forming bacteria in paper and board was the broke (rejected paper or board, which is repulped and recycled into the process) . Gram-negative bacteria were rare in paper and board in spite of their abundance in the stock . A strain of B . pumilus forming clumping, hairy spores may be of significance in aseptic packaging. Infect Immun, 1991 Aug, 59(8), 2567 - 72 Effect of mycobacteria on sensitivity to the cytotoxic effects of tumor necrosis factor; Filley EA et al.; Unlike Mycobacterium leprae, Mycobacterium tuberculosis is not found inside cells other than macrophages and polymorphonuclear cells in vivo, yet previous work has revealed that in vitro it readily enters all cell lines tested . Moreover, these cells are not killed by the intracellular mycobacteria . We report here that when fibroblasts take up live (but not killed) M . tuberculosis H37Rv, they develop greatly increased sensitivity to the toxic effects of tumor necrosis factor (TNF) whether the cell line is inherently sensitive to TNF or not . Ultrasonically disrupted M . tuberculosis also has this property . The increased sensitivity is seen in the absence of metabolic inhibitors, although addition of emetine, an inhibitor of protein synthesis, causes the effect to manifest itself earlier and at a lower concentration of TNF . In contrast, infection with Mycobacterium bovis bacillus Calmette-Guerin induces little or no increased sensitivity to TNF, whereas Mycobacterium avium and M . tuberculosis H37Ra have intermediate sensitivities . We discuss the possibility that virulent tuberculosis strains produce a factor which distorts the normal protective function of TNF, rendering it toxic to host tissues and leading to the classical immunopathology of tuberculous lesions. J Egypt Soc Parasitol, 1991 Aug, 21(2), 575 - 83 Effect of nutritive elements on the extracellular protein of different Bacillus strains, toxic to mosquito larvae; Rady MH et al.; The effect of 5 nutritional elements (glucose, peptone, yeast extract, beef extract and lactic acid) on the growth of the 4 tested Bacillus strains (Bacillus thuringiensis Bactimos 78, B . thuringiensis IPS 82, B . sphaericus Solvay 85 and B . sphaericus Rb 80) as well as on the concentration of their extracellular proteins were studied . The nutritive elements were individually tested either dissolved in distilled water or in a buffer base . Results indicated that the bacterial growth was higher when the nutritive elements were dissolved in a buffer base than in dist . water . The concentration of the extracellular proteins were always higher in the absence of the inorganic minerals, although the toxicity of these extracellular proteins against 3rd instar Culex pipiens larvae increased in the presence of these inorganic elements. J Bacteriol, 1991 Aug, 173(15), 4889 - 92 Temperature-induced protein synthesis in Bacillus stearothermophilus NUB36; Wu L et al.; Cultures of Bacillus stearothermophilus subjected to a temperature shift-up or shift-down of 15 degrees C within the normal temperature range of growth (45 to 65 degrees C) enter a transient adaptation period before exponential growth at the new temperature . The de novo synthesis of some proteins coincides with the adaptation period. J Bacteriol, 1991 Aug, 173(15), 4611 - 7 The Aeromonas hydrophila cphA gene: molecular heterogeneity among class B metallo-beta-lactamases; Massidda O et al.; An Aeromonas hydrophila gene, named cphA, coding for a carbapenem-hydrolyzing metallo-beta-lactamase, was cloned in Escherichia coli by screening an Aeromonas genomic library for clones able to grow on imipenem-containing medium . From sequencing data, the cloned cphA gene appeared able to code for a polypeptide of 254 amino acids whose sequence includes a potential N-terminal leader sequence for targeting the protein to the periplasmic space . These data were in agreement with the molecular mass of the original Aeromonas enzyme and of the recombinant enzyme produced in E . coli, evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of crude beta-lactamase preparations followed by renaturation treatment for proteins separated in the gel and localization of protein bands showing carbapenem-hydrolyzing beta-lactamase activity by a modified iodometric technique . The deduced amino acid sequence of the CphA enzyme showed regions of partial homology with both the beta-lactamase II of Bacillus cereus and the CfiA beta-lactamase of Bacteroides fragilis . Sequence homologies were more pronounced in the regions encompassing the amino acid residues known in the enzyme of B . cereus to function as ligand-binding residues for the metal cofactor . The CphA enzyme, however, appeared to share a lower degree of similarity with the two other enzymes, which, in turn, seemed more closely related to each other . These results, therefore, suggest the existence of at least two molecular subclasses within molecular class B metallo-beta-lactamases. Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 1991 Aug, 13(4), 261 - 6 {Biological assay, reliability test, error analysis and pooled calculation for assessing leucomycin yield}; Tan S; A biological assay for determining leucomycin yield was established by using a curillinear protocol (Sarcina lutea) and (2.2) dosage or (3.3) dosage protocol (Bacillus subtillis 63501) . At the same time, according to the principles of biostatistics and the methods of variance analysis, reliability tests, error analysis and pooled calculations were carried out, so as to hold the confidence limit below the internationally accepted level of 5%. Chem Pharm Bull (Tokyo), 1991 Aug, 39(8), 2063 - 7 Action of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis is significantly influenced by coexisting lipids in substrate-detergent micelles; Kume T et al.; The effects of phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelin (SM), and cholesterol on the activity of phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus thuringiensis were studied in detail in phosphatidylinositol (PI)/detergent mixed micelles . By addition of PC, the enzymatic hydrolysis of PI was significantly stimulated in PI/Triton X-100 as well as PI/sodium deoxycholate (SDC) mixed micelles . SM stimulated enzyme activity toward PI/Triton X-100 micelles at a lower molar ratio of SM to PI, but was rather inhibitory at a ratio higher than 2.0 . The enzyme activity became significantly lower with an increase of PE or cholesterol in PI/Triton X-100 micelles . Actually, both PE and cholesterol were intensively inhibitory when added at a higher molar ratio to PI in Triton X-100-containing micelles . In the system of PI/SDC mixed micelles, not only PC but also SM, PE and cholesterol enhanced the enzymatic hydrolysis of PI . The difference between PI/Triton X-100 and PI/SDC micelles regarding the effects of these lipids on PI-PLC action, must be dependent on the physical state of micelles formed by these detergents and lipids. J Comp Pathol, 1991 Aug, 105(2), 167 - 73 Tyzzer's disease (Bacillus piliformis) in Australian marsupials; Canfield PJ et al.; Tyzzer's disease (Bacillus piliformis infection) was diagnosed in nine marsupials (six possums, a koala, a wombat and a dasyurid) . All but two of the possums were captive . Five of the seven marsupials, for which ages were recorded, were juvenile . Affected marsupials were either found dead or were showing non-specific illness for up to two days before death . Affected livers and hearts showed gross haemorrhage and scattered areas of white discolouration . Microscopically, areas of coagulative necrosis were often associated with neutrophilic infiltrates . Intracellular, slender, faintly basophilic rods were occasionally detected in HE-stained sections . Rods were shown by silver staining techniques to be associated with the lesions in all affected animals . These rods showed a beaded appearance when stained with methenamine silver and were consistently in packets within cells, on the peripheries of lesions and scattered sparsely and irregularly within the central necrotic regions. Appl Environ Microbiol, 1991 Aug, 57(8), 2277 - 82 Toxicity of Bacillus thuringiensis to laboratory populations of the olive fruit fly (Dacus oleae); Karamanlidou G et al.; A survey of Bacillus thuringiensis recovered from the environments of olive groves in Greece was carried out . Of 80 soil samples, 24 were found to contain B . thuringiensis with parasporal crystal inclusions; these were tested for toxicity against the olive fruit fly (Dacus oleae) . Mortality levels of larvae caused by the different isolates varied from 7 to 87% . Higher levels of mortality were observed if a mixture of relatively pure crystals and spores was used compared with the mortality resulting from either fraction alone . We were able to show that the toxicity of the most active isolate is likely to be specific for D . oleae. J Biochem (Tokyo), 1991 Aug, 110(2), 279 - 83 Thermostable alanine racemase of Bacillus stearothermophilus: subunit dissociation and unfolding; Toyama H et al.; The guanidine hydrochloride-induced subunit dissociation and unfolding of thermostable alanine racemase from Bacillus stearothermophilus have been studied by circular dichroism, fluorescence and absorption spectroscopies, and