Curr Microbiol, 2003 Oct, 47(4), 295 - 9 Cloning and expression of the insecticidal crystal protein gene Cry1Ca9 of Bacillus thuringiensis G10-01A from Taiwan granaries; Kao SS et al.; A new cry gene (cry1Ca9) was cloned and sequenced from a Bacillus thuringiensis isolate native to Taiwan (G10-01A) . The cry1C-type gene, designated cry1Ca9, consisted of an open reading frame of 3,567 bp, encoding a protein of 1,189 amino acid residues . The polypeptide has the deduced amino acid sequences predicting molecular masses of 134.7 kDa . The gene sequence was compared against the GenBank nucleotide sequence data base . It was found that the cry1Ca9 gene coded for a 134.7-kDa protoxin which had greater than 99.8% homology with the previously reported cry1Ca1 gene, as only three mismatches were found between the two amino acid sequences . When the Cry1Ca9 toxin was expressed in a crystal-negative strain of B . thuringiensis (cryB-), elliptical crystals were produced . Cell extracts from this recombinant strain appear to have high insecticidal activity against lepidopteran larvae (Plutella xylostella). J Food Prot, 2003 Nov, 66(11), 2070 - 5 Heat resistance of Bacillus spores when adhered to stainless steel and its relationship to spore hydrophobicity; Simmonds P et al.; Twenty-one strains of Bacillus (10 B . stearothermophilus, 3 B . cereus, and 8 B . licheniformis strains) were assayed for spore surface hydrophobicity on the basis of three measures: contact angle measurement (CAM), microbial adhesion to hydrocarbons (MATH), and hydrophobic interaction chromatography (HIC) . On the basis of the spore surface characteristics obtained from these assays, along with data on the heat resistance of these spores in water, eight strains of Bacillus (three B . stearothermophilus, three B . cereus, and two B . licheniformis strains) either suspended in water or adhering to stainless steel were exposed to sublethal heat treatments at 90 to 110 degrees C to determine heat resistance (D-value) . Significant increases in heat resistance (ranging from 3 to 400%) were observed for the eight strains adhering to stainless steel . No significant correlation was found between these heat resistance increases and spore surface characteristics as determined by the three hydrophobicity assays . There was a significant positive correlation between the hydrophobicity data obtained by the MATH assay and those obtained by the HIC assay, but these data did not correlate with those obtained by the CAM assay. Biofouling, 2003 Aug, 19(4), 257 - 67 Microfouling studies on experimental test blocks of steel-making slag and concrete exposed to seawater off Chiba, Japan; Nandakumar K et al.; Microfouling studies with the emphasis on microalgae (Bacillariophyceae) were carried out on test blocks of steel-making slag in comparison with concrete . Two types of slag test blocks, with and without fly-ash as an additional source of silica, and concrete test blocks of size 75 x 26 x 26 mm were used to study microfouling build-up for a period of 30 d, with intermittent samplings after 1, 2, 3, 7, 14 and 21 d . The species composition, cell density, biomass and surface pH of the test pieces were determined, in addition to the hydrographic parameters of the water column . Microfouling studies showed higher numbers of algal species as well as a greater cell density on the slag than on the concrete blocks . This was true with respect to biomass measured as dry weight also . Colonization was significantly delayed in the case of concrete . Navicula spp . and Nitzschia spp . were the initial colonizers on all three types of substrata and were the dominant genera throughout the study period . While the number of species increased, several disappeared after colonization, as a part of community build-up . The surface pH of the slag blocks was near neutral, whilst that of the concrete was highly alkaline during the initial period of exposure . This alkaline surface reduced the rate of species colonization on the concrete blocks initially . The study showed severe biofouling on the slag blocks compared to concrete and thus they were considered an environmentally benign construction material for land protection . The use of slag as the construction material for land protection would greatly reduce the expense compared to concrete. Biotechnol Lett, 2003 Oct, 25(20), 1747 - 50 Biodegradation of exploded cotton stalk by Bacillus sp; Zheng L et al.; The exploded bast, branch and stem of cotton stalk were degraded by alkalophilic Bacillus NT-19, with weight losses of 24%, 20% and 14%, respectively, after 14 d . Compared with a white-rot fungus (Phanerochaete chrysosporium), Bacillus NT- 19 preferentially degraded the non-cellulose components of cotton stem . The relative degree of crystallinity of bast fibers decreased by 8% and the middle lamella was partially removed from the fiber bundle by the Bacillus. Biotechnol Lett, 2003 Oct, 25(20), 1709 - 12 Antifreeze activities of poly(gamma-glutamic acid) produced by Bacillus licheniformis; Shih IL et al.; Various enantiomeric isomers, metals salts and molecular sizes of poly(gamma-glutamic acid), gamma-PGA, produced by Bacillus licheniformis CCRC 12826, were prepared and their antifreeze activities were studied by differential scanning calorimetry . The antifreeze activity of gamma-PGA increased as its molecular weight decreased but was indifferent to its D/L-glutamate composition . The antifreeze activity was cation dependent decreasing in the order Mg2+ >> Ca2+ approximately Na+ >> K+ which follows that of inorganic chlorides in that high ionic charge leads to high antifreeze activity . The mechanism by which the cryoprotective effects of gamma-PGA can be explained is still yet to be determined. FEMS Immunol Med Microbiol, 2003 Nov 28, 39(2), 173 - 80 Mycobacterium bovis bacillus Calmette-Guérin infection promotes SOCS induction and inhibits IFN-gamma-stimulated JAK/STAT signaling in J774 macrophages; Imai K et al.; The resurgence in mycobacterial infection worldwide has led to renewed attention to the pathogenesis of Mycobacterium species . Although interferon-gamma (IFN-gamma) is a principal mediator of macrophage activation, macrophages infected with Mycobacterium are poor in response at the cytokine . However, the molecular mechanisms underlying mycobacterial infection remain unclear . The purpose of this study was to elucidate the mechanism of the poor response to IFN-gamma in mycobacterial infection . Our data clearly demonstrate that this is due to induction of suppressor of cytokine signal (SOCS) negative regulators of IFN-gamma signal transduction that closely correlates with the inhibition of JAK/STAT signaling and gene expression stimulated by IFN-gamma . Mycobacterium bovis bacillus Calmette-Guerin infection induces the production of SOCS-1 and SOCS-3 in murine J774 macrophages . The level of SOCS-1 mRNA increased 1 h and reached a maximum 3 h after the addition of the bacteria . SOCS-3 mRNA expression appeared as early as 1 h after the infection . We also observed that trehalose 6,6'-dimycolate/cord factor, a major component of the Mycobacterium tuberculosis cell wall, induces expression of SOCS and inhibits IFN-gamma-stimulated phosphorylation of STAT1 extensively in the cells . The results in this study suggest that a molecular mechanism of mycobacterial infection affects the unresponsiveness to IFN-gamma in the subsequent growth and spread of macrophages. Tuberculosis (Edinb), 2003, 83(6), 351 - 9 Catalase-peroxidase activity has no influence on virulence in a murine model of tuberculosis; Cardona PJ et al.; The capacity to generate a chronic and persistent infection in the experimental murine model of tuberculosis induced aerogenically by a low-dose inoculum was determined in eight isoniazid-resistant clinical strains of Mycobacterium tuberculosis showing different catalase-peroxidase (C-P) activities . Determination of bacillary concentration in lung and spleen and the percentage of pulmonary parenchyma occupied by granulomas were monitored . Data showed no relation between the lack of C-P activity and the ability to develop a persistent infection, highlighting the potential of C-P negative strains to spread through the community. Biotechnol Adv, 2003 Dec, 22(1-2), 45 - 69 Sustainability of insect resistance management strategies for transgenic Bt corn; Glaser JA et al.; Increasing interest in the responsible management of technology in the industrial and agricultural sectors of the economy has been met thorough the development of broadly applicable tools to assess the "sustainability" of new technologies . An arena ripe for application of such analysis is the deployment of transgenic crops . The new transgenic pesticidal or plant-incorporated protectant (PIP) crops have seen widespread application in the United States based on the features of higher yield, lower applications of insecticides, and control of mycotoxin content . However, open rejection of these new crops in Europe and in other countries has been a surprising message and has limited their worldwide acceptance . The US Environmental Protection Agency's (USEPA) Office of Pesticide Programs (OPP) has worked on the development and analysis of insect resistance management (IRM) strategies and has mandated specific IRM requirements for Bacillus thuringiensis (Bt) crops since 1995 under the Food, Fungicide, Insecticide, and Rodenticide Act . Improvement of data quality and sustainability of IRM strategies have been targeted in an ongoing partnership between the USEPA Office of Research and Development and the Office of Pesticide Programs that will further enhance the agency's ability to develop sustainable insect resistance management strategies for transgenic field corn (Bt corn) producing B . thuringiensis (Bt) insecticidal proteins. Eur J Biochem, 2003 Nov, 270(22), 4488 - 96 Interactions of the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus; Jung HI et al.; The enzymes pyruvate decarboxylase (E1) and dihydrolipoyl dehydrogenase (E3) bind tightly but in a mutually exclusive manner to the peripheral subunit-binding domain (PSBD) of dihydrolipoyl acetyltransferase in the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus . The use of directed mutagenesis, surface plasmon resonance detection and isothermal titration microcalorimetry revealed that several positively charged residues of the PSBD, most notably Arg135, play an important part in the interaction with both E1 and E3, whereas Met131 makes a significant contribution to the binding of E1 only . This indicates that the binding sites for E1 and E3 on the PSBD are overlapping but probably significantly different, and that additional hydrophobic interactions may be involved in binding E1 compared with E3 . Arg135 of the PSBD was also replaced with cysteine (R135C), which was then modified chemically by alkylation with increasingly large aliphatic groups (R135C -methyl, -ethyl, -propyl and -butyl) . The pattern of changes in the values of DeltaG degrees, DeltaH degrees and TDeltaS degrees that were found to accompany the interaction with the variant PSBDs differed between E1 and E3 despite the similarities in the free energies of their binding to the wild-type . The importance of a positive charge on the side-chain at position 135 for the interaction of the PSBD with E3 and E1 was apparent, although lysine was found to be an imperfect substitute for arginine . The results offer further evidence of entropy-enthalpy compensation ('thermodynamic homeostasis') - a feature of systems involving a multiplicity of weak interactions. J Environ Health, 2003 Nov, 66(4), 16 - 21 Decontamination of Bacillus thuringiensis spores on selected surfaces by chlorine dioxide gas; Han Y et al.; This work examined the efficacy of chlorine dioxide (ClO2) gas for the decontamination of Bacillus thuringiensis spores on paper, wood, epoxy, and plastic surfaces . Spores representing an inoculation level of approximately 6 log colony-forming units (CFU) per surface were treated with 5, 10, 15, 20, 25, or 30 milligrams per liter (mg/L) ClO2 gas for 12 hours under 85-92 percent relative humidity and at 22 +/- 1 degrees C . Under the tested treatment conditions, the highest population of surviving spores was found on the paper surface and the lowest was found on the plastic surface (p < .05) . The 5 mg/L ClO2 gas treatment inactivated 2.5, 3.6, 4.0, and 4.9 log spores per surface on paper, wood, epoxy, and plastic surfaces, respectively . A greater than 5-log reduction of spores was achieved on each surface after the 15 mg/L ClO2 gas treatment . The minimum ClO2 gas concentration needed to completely inactivate the inoculated spores was 30 mg/L for paper and wood surfaces, 25 mg/L for epoxy surfaces, and 20 mg/L for plastic surfaces . The results of this study may provide insight into the parameters of effective decontamination procedures for Bacillus spores. Kekkaku, 2003 Oct, 78(10), 653 - 9 {Role of oral fluoroquinolones in patients with respiratory diseases}; Ishida T; The characteristics of recently developed oral fluoroquinolones include their broad spectrum involving gram-positive/gram-negative bacteria and atypical pathogens, potent antimicrobial activity against Pneumococcus, rapid tissue/sputum transfer, prolonged half-life, and reduction of their interaction with other agents . However, it has been reported that the common use of oral fluoroquinolones increases the number of fluoroquinolone-resistant bacterial strains . We review the appropriate use of these agents in patients with respiratory infections . In most cases, upper respiratory inflammation is a viral infection . Generally, antimicrobial agents are not necessary, and should not be administered . In Japan, a large number of antimicrobial agents, especially quinolones, are frequently prescribed to treat upper respiratory infection . This tendency must not be corrected . With respect to treatment for community-acquired pneumonia, it is controversial whether oral fluoroquinolones should be prescribed under various guidelines . In elderly patients and those with an underlying disease, oral fluoroquinolones may be a first-choice treatment at the outpatient clinic, because it is difficult to differentiate atypical pneumonia from bacterial pneumonia, and because the risk of drug-resistant Pneumococcus or gram-negative bacteria is high . With respect to treatment for hospital-acquired pneumonia, oral fluoroquinolones are recommended for patients with moderate or mild conditions without risk factors under the Guidelines established by the Japanese Respiratory Society . Bacteria causing acute infectious exacerbation in patients with chronic pulmonary diseases include gram-positive/gram-negative bacteria and anaerobic bacteria . Therefore, oral fluoroquinolones may be the most appropriate treatment for such patients . New oral fluoroquinolones show potent antimicrobial activity against tubercle bacillus, and may also be effective for infection with bacteria resistant to standard antitubercular agents . It may be controversial whether these agents should be indicated for atypical acid-fast bacterial infection. Pest Manag Sci, 2003 Nov, 59(11), 1197 - 202 Inheritance of resistance to Bt canola in a field-derived population of Plutella xylostella; Sayyed AH et al.; Crops expressing Bacillus thuringiensis (Bt) insecticidal Cry proteins are grown on millions of hectares . Recommendations to delay resistance are based on a high expression/refugia strategy that aims to kill resistant heterozygotes and enable some susceptible insects to survive . Leaf-dip bioassays on F1 crosses of Malaysian populations of diamondback moth (Plutella xylostella (L)) showed that CrylAc resistance was not fully recessive . The survival of ca 50% of heterozygotes on Bt canola (Brassica napus L) leaves expressing low concentrations of CrylAc agreed with a non-fully-recessive model for resistance . Extrapolations based on log dose-logit mortality regressions for heterozygotes using leaf-dip bioassays showed that a relatively high level of expression, of ca 2000 ng CrylAc mg(-1) total leaf protein, would be required to give 90% mortality to heterozygotes . If high enough levels of expression of Bt toxin to kill heterozygotes cannot be achieved and maintained under field conditions, the effectiveness of the high-dose/refugia strategy would be reduced. Sichuan Da Xue Xue Bao Yi Xue Ban, 2003 Oct, 34(4), 614 - 7 {Preliminary study of the molecular regulation of BCG-mediated enhancement of hBD-1 gene expression in human pulmonary gland epithelial cells}; Zhu B et al.; OBJECTIVE: To define the regulation mechanism of the enhancement expression of human beta-defensin-1 (hBD-1) stimulated by bacille Calmette-Guerin (BCG) at transcription level . METHODS: A series of 5'-flanking deletions of hBD-1 gene were ligated into pEGFP-1 and pCAT basic vector . pEGFP hBD-1 recombinants were transfected into SPC-A-1 cells and the green fluorescence protein (GFP) expression was observed by fluorescence microscopy . SPC-A-1 cells were co-transfected with pCAT hBD-1 constructs and pSV-beta-Galactosidase control vector and were stimulated with effective BCG cell wall components . CAT and beta-Gal protein expressions were determined by ELISA . RESULTS: The promoting activity of the -69 region was lower than that of the region -575 and -314 . After being stimulated with the active fraction of BCG cell wall proteins, the relative CAT expression of pCAT hBD-1/-69 still showed the increasing tendency of -2161 construct . Computer consensus match analysis indicated that the nucleotides between -63 bp to -50 bp was the potential binding site for C/EBP beta . CONCLUSION: The nucleotides from -69 to bp is essential to the enhancement of hBD-1 gene transcription by BCG. Bioprocess Biosyst Eng, 2003 Dec, 26(2), 123 - 32 Epub 2003 Nov 14. Monte Carlo simulation of the alpha-amylolysis of amylopectin potato starch . 2 . alpha-amylolysis of amylopectin; Marchal LM et al.; A model is presented that describes all the saccharides that are produced during the hydrolysis of starch by an alpha-amylase . Potato amylopectin, the substrate of the hydrolysis reaction, was modeled in a computer matrix . The four different subsite maps presented in literature for alpha-amylase originating from Bacillus amyloliquefaciens were used to describe the hydrolysis reaction in a Monte Carlo simulation . The saccharide composition predicted by the model was evaluated with experimental values . Overall, the model predictions were acceptable, but no single subsite map gave the best predictions for all saccharides produced . The influence of an alpha(1-->6) linkage on the rate of hydrolysis of nearby alpha(1-->4) linkages by the alpha-amylase was evaluated using various inhibition constants . For all the subsites considered the use of inhibition constants led to an improvement in the predictions (a decrease of residual sum of squares), indicating the validity of inhibition constants as such . As without inhibition constants, no single subsite map gave the best fit for all saccharides . The possibility of generating a hypothetical subsite map by fitting was therefore investigated . With a genetic algorithm it was possible to construct hypothetical subsite maps (with inhibition constants) that gave further improvements in the average prediction for all saccharides . The advantage of this type of modeling over a regular fit is the additional information about all the saccharides produced during hydrolysis, including the ones that are difficult to measure experimentally. J Bacteriol, 2003 Dec, 185(23), 6985 - 9 The Bacillus thuringiensis linear double-stranded DNA phage Bam35, which is highly similar to the Bacillus cereus linear plasmid pBClin15, has a prophage state; Stromsten NJ et al.; Bam35, a 15-kbp double-stranded DNA phage, infects Bacillus thuringiensis . Recently, sequencing of the related Bacillus cereus revealed a 15.1-kbp linear plasmid, pBClin15 . We show that pBClin15 closely resembles Bam35 and demonstrate conversion of Bam35 to a prophage . This state is common, as several B . thuringiensis strains release Bam35-related viruses. Mol Microbiol, 2003 Nov, 50(3), 751 - 62 Disruption of mptpB impairs the ability of Mycobacterium tuberculosis to survive in guinea pigs; Singh R et al.; Protein tyrosine kinases and tyrosine phosphatases from several bacterial pathogens have been shown to act as virulence factors by modulating the phosphorylation and dephosphorylation of host proteins . The identification and characterization of two tyrosine phosphatases namely MptpA and MptpB from Mycobacterium tuberculosis has been reported earlier . MptpB is secreted by M . tuberculosis into extracellular mileu and exhibits a pH optimum of 5.6, similar to the pH of the lysosomal compartment of the cell . To determine the role of MptpB in the pathogenesis of M . tuberculosis, we constructed a mptpB mutant strain by homologous recombination and compared the ability of parent and the mutant strain to survive intracellularly . We show that disruption of the mptpB gene impairs the ability of the mutant strain to survive in activated macrophages and guinea pigs but not in resting macrophages suggesting the importance of its role in the host-pathogen interaction . Infection of guinea pigs with the mutant strain resulted in a 70-fold reduction in the bacillary load of spleens in infected animals as compared with the bacillary load in animals infected with the parental strain . Upon reintroduction of the mptpB gene into the mutant strain, the complemented strain was able to establish infection and survive in guinea pigs at rates comparable to the parental strain . These observations demonstrate a role of MptpB in the pathogenesis of M . tuberculosis. Clin Exp Allergy, 2003 Nov, 33(11), 1512 - 7 Age at bacille Calmette-Guérin vaccination and risk of allergy and asthma; Bager P et al.; BACKGROUND: It has been proposed that early age at bacille Calmette-Guerin (BCG) vaccination protects against the development of allergy . OBJECTIVE: To study whether early age at BCG vaccination was associated with a decreased risk of atopy, allergic rhinitis, and asthma compared to BCG vaccination at later ages in childhood . METHODS: The occurrence of atopy, allergic rhinitis, and asthma was studied in nearly 2000 women participating in the Danish National Birth Cohort study . Detailed information on age at BCG vaccination (age 0-15 years) was available from school health records . Atopic status was assessed serologically by a specific response to 11 common inhalant allergens using serum samples obtained from the women during the period 1997-2001 . Information on allergic rhinitis and asthma was available from telephone interviews . RESULTS: Approximately 85% of the women had been BCG-vaccinated . Age at BCG vaccination was not associated with risk of atopy, allergic rhinitis, or asthma . The odds ratio of atopy, allergic rhinitis, and asthma associated with being vaccinated during the first year of life was 1.05 (95% CI 0.71-1.56), 1.42 (95% CI 0.85-2.36), and 1.71 (95% CI 0.91-3.20), respectively, compared with being vaccinated at the age of 7 years . Adjustment for birth cohort, sibship size, age of the woman's mother at birth, and social class in childhood did not affect the results . CONCLUSION: Our findings suggest that age at BCG vaccination in childhood does not influence the development of allergy or asthma. Clin Exp Immunol, 2003 Nov, 134(2), 303 - 8 Immune response measured in human volunteers vaccinated with autoclaved Leishmania major vaccine mixed with low dose of BCG; Mahmoodi M et al.; The immune responses induced against Leishmania antigens in volunteers who were vaccinated in a double-blind, randomized field efficacy trial of a preparation of autoclaved Leishmania major (ALM) mixed with a low dose of Bacille Calmette-Guerin vaccine (BCG) who developed either a cutaneous leishmaniasis (CL) lesion due to exposure to infected sandfly bite(s) or did not develop a lesion during the course of the trial were studied and compared with those of non-vaccinated controls . Blood samples were also assayed from different groups including volunteers with history of CL and volunteers with previous positive or negative leishmanin skin test (LST) without a history of CL . The vaccinated volunteers had received a single dose of either ALM mixed with a low dose of BCG or the same dose of BCG alone . The LST and in vitro proliferative response (stimulation index, SI), interferon gamma (IFN-gamma) production and, in a few cases, interleukin (IL)-4 production of peripheral blood mononuclear cells to soluble Leishmania antigens were measured . The results indicated that volunteers who developed CL in the vaccine arm showed a slightly higher SI than cases who received BCG alone . Volunteers with history of CL and volunteers with positive LST demonstrated the strongest proliferation indices and IFN-gamma production . The data suggest that a single dose of ALM + BCG induces a weak Th1 response in vaccinated volunteers that is far lower than that in volunteers with prior subclinical infection or volunteers with history of CL, who are presumed to be immune. Clin Exp Immunol, 2003 Nov, 134(2), 285 - 94 Interferon-gamma and skin test responses of schoolchildren in southeast England to purified protein derivatives from Mycobacterium tuberculosis and other species of mycobacteria; Weir RE et al.; The immune responses of schoolchildren in southeast England to Mycobacterium tuberculosis and other species of mycobacteria were studied prior to vaccination with bacille Calmette-Guerin (BCG) . Data are presented for tuberculin (Heaf) skin test and interferon-gamma (IFN-gamma) responses to M . tuberculosis purified protein derivative (PPD), and IFN-gamma responses to PPDs from eight other environmental mycobacteria, measured in 424 schoolchildren (13-15 years of age) . Responses to M . tuberculosis PPD were detected in 27% of schoolchildren by in vitro IFN-gamma response and in 20% by the Heaf test . IFN-gamma responses were more prevalent to PPDs from species of mycobacteria other than M . tuberculosis, predominantly those of the MAIS complex and M . marinum (45-60% responders) . Heaf test and IFN-gamma responses were associated (P<0.001) for M . tuberculosis, MAIS and M . marinum . These findings have implications for appropriate implementation of vaccination against tuberculosis. Clin Microbiol Infect, 2003 Sep, 9(9), 973 - 5 Nosocomial bacteremia and catheter infection by Bacillus cereus in an immunocompetent patient; Hernaiz C et al.; We present a case of Bacillus cereus bacteremia and catheter infection in an immunocompetent patient subjected to abdominal surgery, who recovered following central catheter removal and treatment with piperacillin/tazobactam. Allergy, 2003 Nov, 58(11), 1114 - 6 Effects of BCG revaccination on asthma; Choi IS et al.; In a study conducted 1 year ago, we found that Th1 immune enhancement following Bacille Calmette-Guerin (BCG) vaccination effectively suppressed human asthma . To investigate whether revaccination would further improve lung function, BCG vaccine was given again . Current lung function tended to improve in the Repeated BCG group (n = 9), but not in the Single BCG group (previously the placebo group) (n = 11), compared with that 1 year ago . The BCG vaccination improved lung function in both groups, and the Repeated BCG group showed a significant increase in the peripheral blood interferon gamma/interleukin 4 ratio . These findings suggest that repeated BCG vaccinations might be effective in asthma therapy. J Theor Biol, 2003 Dec 21, 225(4), 493 - 6 DNA-membrane interactions can localize bacterial cell center; Rabinovitch A et al.; In actively growing bacterial cells, the DNA exerts stress on the membrane, in addition to the turgor caused by osmotic pressure . This stress is applied through coupled transcription/translation and insertion of membrane proteins (so-called "transertion" process) . In bacillary bacteria, the strength of this interaction varies along cell length with a minimum at its midpoint, and hence can locate the cell's equator for the assembly of the FtsZ-ring. Proc R Soc Lond B Biol Sci, 2003 Nov 7, 270(1530), 2263 - 70 Rapid evolution and the cost of resistance to Bacillus thuringiensis in greenhouse populations of cabbage loopers, Trichoplusia ni; Janmaat AF et al.; The microbial insecticide Bacillus thuringiensis (Bt) has become the mainstay of non-chemical control of Lepidopteran pests, either as sprays or through the incorporation of Bt toxins into transgenic crops . Given the wide use of Bt, it is striking that currently only one pest species, Plutella xylostella, has been reported to have developed significant resistance to Bt outside the laboratory . By contrast, we report here the frequent and rapid development of resistance to B . thuringiensis kurstaki (Dipel, Abbott) in populations of cabbage loopers, Trichoplusia ni, in commercial greenhouses . Resistance to Bt appears to be costly and there is a rapid decline of resistance in populations collected from greenhouses and maintained in the laboratory without selection . Management of pests resistant to Bt in vegetable greenhouses will require sporadic use of Bt-based sprays or alternatively use of sprays containing other Bt toxins. Curr Opin Allergy Clin Immunol, 2003 Dec, 3(6), 481 - 6 Immunotherapy with mycobacteria; Walker C et al.; PURPOSE OF REVIEW: To summarize and evaluate critically recent progress with mycobacteria as a potential novel disease modifying treatment strategy in asthma . RECENT FINDINGS: The link between exposure to pathogenic or saprophytic mycobacteria and protection from allergic diseases is still controversial, and recent epidemiological studies, which addressed only exposure to Mycobacterium tuberculosis or bacillus Calmette-Guerin, did not help to clarify this issue . Moreover, the clear efficacy of mycobacterial treatment seen in animal models has not been reproduced in human asthma, and a recent small study testing the hypothesis that heat-killed Mycobacterium vaccae attenuates asthmatic reactions after allergen challenge did not provide convincing results . However, it has been shown that treatment of mice with M . vaccae induces the generation of allergen-specific T regulatory cells capable of suppressing allergen-mediated eosinophilic lung inflammation, suggesting that a general deficiency of T regulatory cell activity might be responsible for the increased prevalence of asthma . This hypothesis is supported by findings that a lack of T regulatory cells, as found in genetic disorders of man and mouse attributable to a mutation of Foxp3, a transcription factor specifically expressed by T regulatory cells, is associated with manifestations of severe atopy and autoimmunity, precisely the spectrum of diseases linked to the hygiene hypothesis . SUMMARY: Further studies on the relationship between mycobacteria and atopic disorders are needed, but there is reason to believe that the novel findings and molecular mechanisms associated with mycobacterial infections will further strengthen the currently unproved therapeutic value of immunotherapy with mycobacteria. FEMS Microbiol Lett, 2003 Nov 7, 228(1), 21 - 6 CbiX-homologous protein (CbiXhp), a metal-binding protein, from Streptomyces seoulensis is involved in expression of nickel-containing superoxide dismutase; Kim IK et al.; To find the accessory proteins participating in expression and maturation of nickel-containing superoxide dismutase (NiSOD), a metal-binding protein (CbiXhp) homologous to cobaltochelatase (CbiX) of Bacillus megaterium was isolated by nickel-nitrilotriacetic acid resin from Streptomyces seoulensis . The deduced amino acid sequence of cbiXhp showed 87% and 39% identity to CbiX of Streptomyces coelicolor and that of B . megaterium, respectively . Overexpression of CbiXhp increased the activity and the expression of NiSOD in the presence and absence of nickel, but to a lesser extent in its absence . This result indicates that CbiXhp is involved in the expression of NiSOD. FEMS Microbiol Lett, 2003 Nov 7, 228(1), 1 - 9 Genes for an alkaline D-stereospecific endopeptidase and its homolog are located in tandem on Bacillus cereus genome; Komeda H et al.; Alkaline D-peptidase (Adp) from Bacillus cereus DF4-B is a D-stereospecific endopeptidase acting on oligopeptides composed of D-phenylalanine and the primary structure deduced from its gene, adp, shows a similarity with D-stereospecific hydrolases from Ochrobactrum anthropi strains . We have isolated DNA fragments covering the flanking region of adp from DF4-B genome and found an additional gene, adp2, located upstream of adp . The deduced amino acid sequence of Adp2 showed 96% and 85% identity with those of Adp from B . cereus strains AH559 and DF4-B, respectively . The recombinant Adp2 expressed in Escherichia coli was purified to homogeneity and characterized . It had hydrolyzing activity toward (D-Phe)3, (D-Phe)4, and (D-Phe)6 but did not act on (L-Phe)4, D-Phe-NH2, and L-Phe-NH2, some characteristics that are closely related to those of Adp from strain DF4-B . These results indicate that highly homologous genes encoding D-stereospecific endopeptidases are arranged in a tandem manner on the genomic DNA of B . cereus DF4-B. Biochemistry, 2003 Nov 18, 42(45), 13304 - 18 Glycosynthase activity of Bacillus licheniformis 1,3-1,4-beta-glucanase mutants: specificity, kinetics, and mechanism; Faijes M et al.; Glycosynthases are engineered retaining glycosidases devoid of hydrolase activity that efficiently catalyze transglycosylation reactions . The mechanism of the glycosynthase reaction is probed with the E134A mutant of Bacillus licheniformis 1,3-1,4-beta-glucanase . This endo-glycosynthase is regiospecific for formation of a beta-1,4-glycosidic bond with alpha-glycosyl fluoride donors (laminaribiosyl as the minimal donor) and oligosaccharide acceptors containing glucose or xylose on the nonreducing end (aryl monosaccharides or oligosaccharides) . The pH dependence of the glycosynthase activity reflects general base catalysis with a kinetic pK(a) of 5.2 +/- 0.1 . Kinetics of enzyme inactivation by a water-soluble carbodiimide (EDC) are consistent with modification of an active site carboxylate group with a pK(a) of 5.3 +/- 0.2 . The general base is Glu138 (the residue acting as the general acid-base in the parental wild-type enzyme) as probed by preparing the double mutant E134A/E138A . It is devoid of glycosynthase activity, but use of sodium azide as an acceptor not requiring general base catalysis yielded a beta-glycosyl azide product . The pK(a) of Glu138 (kinetic pK(a) on k(cat)/K(M) and pK(a) of EDC inactivation) for the E134A glycosynthase has dropped 1.8 pH units compared to the pK(a) values of the wild type, enabling the same residue to act as a general base in the glycosynthase enzyme . Kinetic parameters of the E134A glycosynthase-catalyzed condensation between Glcbeta4Glcbeta3GlcalphaF (2) as a donor and Glcbeta4Glcbeta-pNP (15) as an acceptor are as follows: k(cat) = 1.7 s(-)(1), K(M)(acceptor) = 11 mM, and K(M)(donor) < 0.3 mM . Donor self-condensation and elongation reactions are kinetically evaluated to establish the conditions for preparative use of the glycosynthase reaction in oligosaccharide synthesis . Yields are 70-90% with aryl monosaccharide and cellobioside acceptors, but 25-55% with laminaribiosides, the lower yields (and lower initial rates) due to competitive inhibition of the beta-1,3-linked disaccharide acceptor for the donor subsites of the enzyme. Rev Esp Salud Publica, 2003 Sep-Oct, 77(5), 553 - 65 {Risk factors for tuberculous disease in AIDS cases reported in Brazil, from 1980 to 2000}; Laguardia J et al.; BACKGROUND: Aids affects the epidemiological characteristics of tuberculosis, due to both the changes in the clinical progress of the jointly affected patients, as well as the increase in the number of tuberculosis cases among individuals having tested positive for hiv/aids . This may lead to an increase in the transmission of the bacillus among the population . this study is aimed at identifying the factors related to the occurrence of tuberculosis among aids cases reported to the brazilian ministry of health . METHODS: a cross-sectional epidemiological study was conducted, the aids cases initially having been classified into 2 categories (1) with tuberculosis and (2) without tuberculosis, a comparison was subsequently drawn among 4 groups: (1) no opportunist infection, (2) only tuberculosis (pulmonary and/or disseminated), (3) with other opportunist infections except tuberculosis and (4) with tuberculosis in conjunction with other opportunist infections . The bivariate analysis was adjusted by logistic regression . RESULTS: In the regression analysis, the highest tuberculosis prevalences were significantly linked to age under 40, little schooling, living in the southeast region of the country and diagnosis made prior to 1996 . The means of transmission most closely associated to the presence of tuberculosis was the use of intravenous drugs, followed by heterosexual relations . when 4 consecutive periods in the evolution of the reporting aids case definition, there had been a decrease in cases with tuberculosis since 1996, as well as significant increases in the proportion of patients with no opportunist infection . CONCLUSIONS: Gender, age and school attendance are predictors of the presence of tuberculosis is among opportunist infections. Nat Biotechnol, 2003 Dec, 21(12), 1493 - 7 Epub 2003 Nov 09. Transgenic plants expressing two Bacillus thuringiensis toxins delay insect resistance evolution; Zhao JZ et al.; Preventing insect pests from developing resistance to Bacillus thuringiensis (Bt) toxins produced by transgenic crops is a major challenge for agriculture . Theoretical models suggest that plants containing two dissimilar Bt toxin genes ('pyramided' plants) have the potential to delay resistance more effectively than single-toxin plants used sequentially or in mosaics . To test these predictions, we developed a unique model system consisting of Bt transgenic broccoli plants and the diamondback moth, Plutella xylostella . We conducted a greenhouse study using an artificial population of diamondback moths carrying genes for resistance to the Bt toxins Cry1Ac and Cry1C at frequencies of about 0.10 and 0.20, respectively . After 24 generations of selection, resistance to pyramided two-gene plants was significantly delayed as compared with resistance to single-gene plants deployed in mosaics, and to Cry1Ac toxin when it was the first used in a sequence . These results have important implications for the development and regulation of transgenic insecticidal plants. Am J Infect Control, 2003 Oct, 31(6), 347 - 53 Comparison of a whole-blood interferon-gamma assay and tuberculin skin testing in patients with active tuberculosis and individuals at high or low risk of Mycobacterium tuberculosis infection; Fietta A et al.; BACKGROUND: QuantiFeron-TB (QIFN) is a whole-blood interferon-;gamma assay for the recognition of cell-mediated immune response to Mycobacterium tuberculosis infection . OBJECTIVES: To compare the QIFN assay with the tuberculin skin test (TST) in patients with newly diagnosed culture-proven tuberculosis (TB) and healthy volunteers with high or low risk of latent M tuberculosis infection and to identify factors associated with discordance between tests . METHOD: Two-hundred fifty-eight subjects underwent both assays . All participants completed a detailed questionnaire, and data from TB patients' medical records were collected . RESULTS: In the entire study population, agreement between tests was moderate and the correlation between the magnitude of QIFN response and the TST induration diameter was significant . In volunteers with no known risk of exposure to M tuberculosis, the specificity of the assays was comparable . However, in subjects with active TB or those vaccinated with bacille Calmette-Guerin, the QIFN assay detected more reactors than did the TST . In these individuals, agreement between assays was poor and no correlation or only a weak correlation was found between the diameter of TST induration and the magnitude of the interferon-gamma responses . CONCLUSIONS: The sensitivity of the QIFN assay is greater than that of the TST in patients with active TB before the initiation of anti-TB chemotherapy, but its specificity is influenced more by bacille Calmette-Guerin vaccination . The QIFN assay may provide an improvement over the current practice of the use of the TST to support diagnosis of active M tuberculosis infection in the clinic; however, QIFN cannot be considered an adequate replacement for the TST in the screening for latent infection. J Biochem (Tokyo), 2003 Oct, 134(4), 543 - 50 Deciphering the molecular basis of the broad substrate specificity of alpha-glucosidase from Bacillus sp . SAM1606; Noguchi A et al.; The alpha-glucosidase of Bacillus sp . strain SAM1606 is a member of glycosyl hydrolase family 13, and shows an extraordinarily broad substrate specificity and is one of very few alpha-glucosidases that can efficiently hydrolyze the alpha-1,1-glucosidic linkage of alpha,alpha'-trehalose (trehalose) . Phylogenetic analysis of family-13 enzymes suggests that SAM1606 alpha-glucosidase may be evolutionally derived from an alpha-1,6-specific ancestor, oligo-1,6-glucosidase (O16G) . Indeed, replacement of Pro(273*) and Thr(342*) of B . cereus O16G by glycine and asparagine (the corresponding residues in the SAM1606 enzyme), respectively, was found to cause 192-fold enhancement of the relative catalytic efficiency for trehalose, suggesting that O16G may easily "evolved" into an enzyme with an extended substrate specificity by substitution of a limited number of amino acids, including that at position 273* (an asterisk indicates the amino-acid numbering of the SAM1606 sequence) . To probe the role of the amino acid at position 273* of alpha-glucosidase in determination of the substrate specificity, the amino acid at position 273 of SAM1606 alpha-glucosidase was replaced by all other naturally occurring amino acids, and the resultant mutants were kinetically characterized . The results showed that substitution of bulky residues (e.g., isoleucine and methionine) for glycine at this position resulted in large increases in the K(m) values for trehalose and maltose, whereas the affinity to isomaltose was only minimally affected by such an amino-acid substitution at this position . Three-dimensional structural models of the enzyme-substrate complexes of the wild-type and mutant SAM1606 alpha-glucosidases were built to explore the mechanism responsible for these observations . It is proposed that substitution by glycine at position 273* could eliminate steric hindrance around subsite +1 that originally occurred in parental O16G and is, at least in part, responsible for the acquired broad substrate specificity of SAM1606 alpha-glucosidase. Biochemistry (Mosc), 2003 Sep, 68(9), 1012 - 9 Expression, isolation, purification, and biochemical properties of trehalose-6-phosphate hydrolase from thermoresistant strain Bacillus sp . GP16; Karelov DV et al.; Here we describe cloning, expression, and purification of the enzyme trehalose-6-phosphate hydrolase from thermoresistant strain Bacillus sp . GP16 . Principal biochemical properties of the enzyme at different pH and temperature values were determined . Entropy and enthalpy of activation of the enzyme for substrates trehalose-6-phosphate and p-nitrophenyl glucoside were calculated, and the dependence of the kinetic parameters from ionic strength was established. Biochemistry (Mosc), 2003 Sep, 68(9), 984 - 7 Cloning and sequencing of the gene of site-specific nickase N.BspD6I; Perevyazova TA et al.; A fragment of chromosomal DNA from Bacillus species D6 containing the gene of nickase N.BspD6I and the regions adjacent to its 5;- and 3;-ends was cloned and sequenced . The nucleotide sequence of the nickase gene, except of one neutral change, is homologous to the nicking endonuclease N.BstNBI gene sequenced by Higgens et al . (2001) . After integration of a PCR-copy of the nickase gene into an expression vector pET28b under the control of the phage T7 promoter, specific nicking activity was detected in the lysates of transformed E . coli cells. Biochemistry (Mosc), 2003 Sep, 68(9), 967 - 75 M.BstF5I-2 and M.BstF5I-4 DNA methyltransferases from BstF5I restriction-modification system of Bacillus stearothermophilus F5; Chernukhin VA et al.; The BstF5I restriction-modification system from Bacillus stearothermophilus F5 includes four site-specific DNA methyltransferases, thus differing from all known restriction-modification systems . Here we demonstrated for the first time that one bacterial cell can possess two pairs of methylases with identical substrate specificities (methylases BstF5I-1 and BstF5I-3 recognize GGATG, whereas methylases BstF5I-2 and BstF5I-4 recognize CATCC) that modify adenine residues on both DNA strands . Different chromatographic methods provide homogenous preparations of methylases BstF5I-2 and BstF5I-4 . We estimated the principal kinetic parameters of the reaction of transfer of methyl group from the donor S-adenosyl-L-methionine to the recognition site 5;-CATCC-3; catalyzed by BstF5I-2 and BstF5I-4 DNA {N6-adenine}-methyltransferases from the BstF5I restriction-modification system. J Clin Microbiol, 2003 Nov, 41(11), 5291 - 3 Pericardial effusion in a homeless man due to Bartonella quintana; Levy PY et al.; Bartonella quintana may cause trench fever, endocarditis, bacillary angiomatosis, and chronic bacteremia, and a reemergence among homeless populations in cities has been noted . Pericarditis from Rickettsia conorii and Coxiella burnetii infection has been described, but there have been no reports of pericarditis due to Bartonella spp . We report a case of pericardial effusion due to Bartonella quintana in a homeless man, diagnosed on the basis of PCR detection of Bartonella quintana in a pericardial biopsy sample and a fourfold rise in antibody titers . The patient recovered within 2 weeks with antibiotics active against bartonellae. Vaccine, 2003 Dec 8, 22(1), 70 - 6 Oral vaccination with Mycobacterium bovis BCG in a lipid formulation induces resistance to pulmonary tuberculosis in brushtail possums; Aldwell FE et al.; A method was developed for formulating Mycobacterium bovis bacille Calmette-Guerin (BCG) for oral vaccination against tuberculosis . Selected lipid-based formulations of BCG were tested in the brushtail possum for their ability to elicit immune responses and protection against bovine tuberculosis . Formulation of BCG in lipid matrices maintained bacteria in a dormant but viable state . Oral delivery of 2 x 10(8) colony forming units of formulated BCG to possums induced strong lymphocyte proliferation responses to bovine purified protein derivative (PPD) in peripheral blood lymphocytes . Oral vaccination of possums also reduced the severity of disease following aerosol challenge with virulent M . bovis compared with animals vaccinated with non-formulated BCG . In a second experiment, levels of protection with lipid-formulated oral BCG were similar to those seen with subcutaneous BCG vaccination . Our data shows that formulated oral BCG is an efficient means of inducing protection against bovine tuberculosis in possums and should be a practical means of vaccinating wildlife against tuberculosis. J Immunol Methods, 2003 Nov, 282(1-2), 45 - 52 Generation of an affinity column for antibody purification by intein-mediated protein ligation; Sun L et al.; Coupling an antigenic peptide to a solid support is a crucial step in the affinity purification of a peptide-specific antibody . Conventional methods for generating reactive agarose, cellulose or other matrices for peptide conjugation are laborious and can result in a significant amount of chemical waste . In this report, we present a novel method for the facile production of a peptide affinity column by employing intein-mediated protein ligation (IPL) in conjunction with chitin affinity chromatography . A reactive thioester was generated at the C-terminal of the chitin binding domain (CBD) from the chitinase A1 of Bacillus circulans WL-2 by thiol-induced cleavage of the peptide bond between the CBD and a modified intein . Peptide epitopes possessing an N-terminal cysteine were ligated to the chitin bound CBD tag . We demonstrate that the resulting peptide columns permit the highly specific and efficient affinity purification of antibodies from animal sera. J Indian Med Assoc, 2003 Mar, 101(3), 198, 200 - 3 Tuberculosis and general practitioners; Tripathy SN; In spite of advancement of knowledge in diagnosis and management, tuberculosis is still the biggest health problem . There are more than 400 million people infected with tuberculous bacillus and more than 14 million cases are suffering from the disease . Prevention, control and care of tuberculosis are possible . General practitioners play a major role in combating the disease . Quick and right diagnosis and treating the cases effectively should be the motto . Sputum microscopy is the backbone of diagnosis of tuberculosis . With the availability of modern chemotherapy the outcome of a tuberculosis case has undergone dramatic changes . Short-course chemotherapy is the standard choice while tackling cases of tuberculosis . Results of directly observed treatment (DOT) strategy are very good provided execution is proper . HIV/AIDS, another dreaded disease if becomes co-infected along with tuberculosis, mortality and morbidity become very high . The general practitioners are the major strength of healthcare system in any society . They should be good enough to know all about the control programmes well implemented in the country. Parasitol Res, 2004 Jan, 92(1), 53 - 7 Epub 2003 Nov 05. Protection of mice infected with Plasmodium berghei by Bacillus thuringiensis crystal proteins; Xu Z et al.; Eight Bacillus thuringiensis strains were used to test their activity against Plasmodium berghei . When crystal proteins extracted from strains 007, 017, 020, 021, 030, 032, and 037 were injected into plasmodium-infected mice through the tail vein at a rate of 0.45-1.5 mg per mouse, the lengths of survival for the mice were extended up to 5 days (from 8.5 days to 13.5-15 days) . Blood-cell staining demonstrated that normal erythrocytes were lightly stained and regularly shaped while the erythrocytes from plasmodia-infected mice swelled, lost shape and even lysed . This means that the crystal proteins could protect erythrocytes from the plasmodium's attack . Proteins analysis revealed that most of the proteins are homologues of classic crystal proteins, with the exception of the 120-kDa protein of strain 020, a surface-layer protein . This study suggested a novel way to control plasmodial infections and even malaria. Microbiology, 2003 Nov, 149(Pt 11), 3213 - 20 Koch's bacillus - a look at the first isolate of Mycobacterium tuberculosis from a modern perspective; Taylor GM et al.; Using molecular methods the authors have studied mycobacterial DNA taken from a 19th century victim of tuberculosis . This was the case from which Robert Koch first isolated and cultured the organism responsible for tuberculosis . The mycobacteria were preserved within five glass culture tubes as abundant bacterial colonies on slopes of a gelatinous culture medium of unknown composition . Originally presented by Koch to surgical laryngologist Walter Jobson Horne in London in 1901, the relic has, since 1983, been in the care of the Royal College of Surgeons of England . Light and electron microscopy established the presence of acid-fast mycobacteria but showed that morphological preservation was generally poor . Eleven different genomic loci were successfully amplified by PCR . This series of experiments confirmed that the organisms were indeed Mycobacterium tuberculosis and further showed that the original strain was in evolutionary terms similar to 'modern' isolates, having undergone the TB D1 deletion . Attempts to determine the genotypic group of the isolate were only partially successful, due in part to the degraded nature of the DNA and possibly also to a truncation in the katG gene, which formed part of the classification scheme . Spoligotyping resulted in amplification of DR spacers consistent with M . tuberculosis but with discrepancies between independent extracts, stressing the limitations of this typing method when applied to poorly preserved material. J Clin Pathol, 2003 Nov, 56(11), 868 - 70 Inflammatory pseudotumour associated with chronic persistent Eikenella corrodens infection: a case report and brief review; Lee SH et al.; Inflammatory pseudotumour is an uncommon mass forming lesion, representing the histological expression of an infective or reactive/reparative process (pseudotumour) in most cases, and a bona fide neoplasm (for example, inflammatory myofibroblastic tumour) in a minority of cases . This report describes the case of an inflammatory pseudotumour with a pathology that unveiled proliferative CD68 positive and actin negative spindle shaped cells, with a mild mixed inflammatory infiltrate, and a culture that yielded an uncommon fastidious bacillus, Eikenella corrodens . The clinical course was indolent but protracted, with insidious progression to multifocal non-contiguous lesions, involving the lungs, liver, spleen, left kidney, and deep neck tissue, all of which responded to medical treatment with appropriate antibiotics . It is of paramount importance that clinicians search for an infective cause of an inflammatory pseudotumour, to ensure appropriate treatment. Toxicol In Vitro, 2003 Oct-Dec, 17(5-6), 745 - 51 A new method for in vitro detection of microbially produced mitochondrial toxins; Hoornstra D et al.; Sperm motility inhibition assay, earlier shown valuable for the detection of food poisoning non-protein toxins of Bacillus species was developed into an assay useful for specific detection of mitochondria damaging toxins . This was done by assessing the dissipation of the mitochondrial inner membrane transmembrane potential, Deltapsim under conditions where the plasma membrane permeability barrier remained intact . The Deltapsim was estimated as the intensity of orange JC-1 fluorescence in the mitochondrial sheath of the exposed spermatozoa . The plasma membrane integrity of the same cells was assessed by observing the exclusion of propidium iodide from the cytoplasm . Three types of mitochondrial toxic responses to microbially made bioactive substances were recognised . Mitochondrial toxicity by gramicidin (A, B, C, D), nigericin, salinomycin, narasin, monensin, calcimycin and antimycin A was characterised by gradual fading of the JC-1 fluorescence in the mitochondria . Dissipation of the Deltapsim by cereulide, valinomycin and enniatin (A, A1, B, B1) was visible as spotwise quenching of the mitochondrial JC-1 fluorescence . In addition these substances caused hyperpolarisation of the plasma membrane . Oligomycin (A, B, C), ionomycin and staurosporine inhibited the spermatozoan motility, but Deltapsim was fully preserved . Surfactin and lichenysin A caused mitochondrial damage at concentrations where the plasma membrane was also damaged. Toxicol In Vitro, 2003 Oct-Dec, 17(5-6), 737 - 44 In vitro assay for human toxicity of cereulide, the emetic mitochondrial toxin produced by food poisoning Bacillus cereus; Jaaskelainen EL et al.; The in vitro boar spermatozoon test was compared with the LC ion trap MS analysis for measuring the cereulide content of a pasta dish, implemented in serious emetic food poisoning caused by Bacillus cereus . Both assays showed that the poisonous food contained approximately 1.6 microg of cereulide g(-1) implying the toxic dose in human as < or =8 microg kg(-1) body weight . The threshold concentration of cereulide provoking visible mitochondrial damage in boar sperm exposed in vitro was 2 ng of cereulide ml(-1) of extended boar sperm . The same threshold value was found for cereulide extracted from the food and from the cultured bacteria . This shows that other constituents of the food did not enhance or mask the effects of cereulide . Exposure of four human cell lines (HeLa, Caco-2, Calu-3 and Paju) to cereulide showed that the threshold concentration for the loss of mitochondrial membrane potential in human cells was similar to that observed in boar sperm . Human cells and boar sperm were equally sensitive to cereulide . The results show that boar spermatozoan assay is useful for detecting cereulide concentrations toxic to humans . Spermatozoa in commercially available extended fresh boar and cryopreserved bull semen were compared, boar sperms were 100 times more sensitive to cereulide than bull sperms. Toxicol In Vitro, 2003 Oct-Dec, 17(5-6), 623 - 8 Tamoxifen induces ultrastructural alterations in membranes of Bacillus Stearothermophilus; Luxo C et al.; Tamoxifen (TAM), a non-steroid antiestrogen, is the mostly used drug for chemotherapy and chemoprevention of breast cancer . However, the mechanisms by which TAM inhibits cell proliferation in breast cancer are not fully understood . TAM strongly incorporates in biomembranes and a variety of effects have been assigned to biophysical and biochemical interactions with membranes . Therefore, a better understanding of the physicochemical basis of interaction of TAM with biomembranes is essential to elucidate the molecular mechanisms of action . A strain of Bacillus stearothermophilus has been used as a model to clarify the interaction of TAM with the cell membrane . TAM effects on the ultrastructure of membranes of this bacterium were evaluated by electron microscopy . Important ultrastructural alterations were observed in B . stearothermophilus treated with TAM, namely change in the geometry of the membrane profile from asymmetric to symmetric, disaggregation of ribosomes, coagulation of the cytoplasmic matrix, occurrence of mesossomes, appearance of fractures in membranes and the alteration of the ultrastructure of cell wall . These ultrastructural alterations confirm that TAM is a membrane-active drug and that membrane damage may be involved in molecular mechanisms of cell death induced by this drug. Rev Clin Esp, 2003 Nov, 203(11), 532 - 5 {High resolution computerized tomography in pulmonary tuberculosis with negative sputum bacilloscopy}; Gonzalez Constan E et al.; CONTEXT: Pulmonary tuberculosis with negative sputum bacilloscopy involves diagnostic difficulties when there are not available liquid culture media due to the slow growth of Mycobacterium tuberculosis in the traditional culture media, and also due to the need for carrying out invasive examinations . OBJECTIVE: Evaluate the high resolution computerized tomography findings (HRCT) in patients with pulmonary tuberculosis and negative bacilloscopy of sputum . METHOD: Prospectively 28 patients (12 with active tuberculosis and 16 with inactive tuberculosis) were evaluated through HRCT, assessing the following findings: centrolobular nodules, multiple branched linear structures, macronodules, cavitation, consolidation, enlargement of interlobular septums, ground-glass change, bronchiectases, emphysema, broncovascular distortion, fibrotic changes, calcified mediastinal adenopathies, parenchymous calcification, pleural enlargement, and pleural effusion . RESULTS: The findings that were associated significantly to the active disease were: consolidation (67%), macronodules (67%) and centrolobular nodules (67%) . The presence of centrolobular nodules and/or consolidation had a sensitivity of 83% and a specificity of 87% . The findings significantly associated to inactive disease were bronchiectases (87%) and broncovascular distortion (62%) . CONCLUSION: Our results support the value of HRCT in patients with pulmonary tuberculosis and negative sputum bacilloscopy, since the finding of centrolobular nodules and/or consolidation has good sensitivity and specificity for the diagnosis of active pulmonary disease. J Med Entomol, 2003 Sep, 40(5), 672 - 7 Susceptibility profile of Culex quinquefasciatus (Diptera: Culicidae) to Bacillus sphaericus on selection with rotation and mixture of B . sphaericus and B . thuringiensis israelensis; Zahiri NS et al.; Several strains of Bacillus sphaericus (Neide) (Bsph), a spore-forming bacterium that produces potent mosquitocidal toxins, have been widely used against mosquito larvae globally . In some locations in the field and in the laboratory, moderate to high levels of resistance in Culex pipiens complex mosquito larvae to Bsph have been reported . Practical strategies for dealing with the resistance problem and more importantly to delay or possibly prevent development of resistance to Bsph in the first place are urgently needed . From published reports, we deduced that Bsph-resistant mosquito larvae are completely susceptible to Bacillus thuringiensis ssp . isreaelensis (Bti) and that there is no appearance of cross-resistance to Bti . On the basis of this information, we hypothesized that Bti offers a good potential for the management of resistance to Bsph, especially when it comes to delaying or preventing emergence of resistance . We conducted laboratory studies to see whether rotation of Bsph and Bti or mixture of both delays or prevents resistance development in Bsph-susceptible California Cx . quinquefasciatus . Development of resistance using Bsph alone increased after F15 and fluctuated slightly on further selections . Rotation of Bsph and Bti resulted in much higher level and rapid emergence of resistance to Bsph . However, selection with mixtures of Bti and Bsph for 36 generations showed no emergence of resistance . On the basis of this information, we believe mixtures have a good potential in delaying or preventing Bsph resistance. J Dairy Sci, 2003 Oct, 86(10), 3075 - 81 Inactivation of spores of Bacillus cereus in cheese by high hydrostatic pressure with the addition of nisin or lysozyme; Lopez-Pedemonte TJ et al.; The objective of this work was to study high hydrostatic pressure (HHP) inactivation of spores of Bacillus cereus ATCC 9139 inoculated in model cheeses made of raw milk, together with the effects of the addition of nisin or lysozyme . The concentration of spores in model cheeses was approximately 6-log10 cfu/g of cheese . Cheeses were vacuum packed and stored at 8 degrees C . All samples except controls were submitted to a germination cycle of 60 MPa at 30 degrees C for 210 min, to a vegetative cells destruction cycle of 300 or 400 MPa at 30 degrees C for 15 min, or to both treatments . Bacillus cereus counts were measured 24 h and 15 d after HHP treatment . The combination of both cycles improved the efficiency of the whole treatment . When the second pressure-cycle was of 400 MPa, the highest inactivation (2.4 +/- 0.1 log10 cfu/g) was obtained with the presence of nisin (1.56 mg/L of milk), whereas lysozyme (22.4 mg/L of milk) did not increase sensitivity of the spores to HHP . For nisin (0.05 and 1.56 mg/L of milk), no significant differences were found between counts at 24 h and 15 d after treatment . Considering that mesophilic spore counts usually range from 2.6 to 3.0 log10 cfu/ml in raw milk, HHP at mild temperatures with the addition of nisin may be useful for improving safety and preservation of soft curd cheeses made from raw milk. Prikl Biokhim Mikrobiol, 2003 Sep-Oct, 39(5), 536 - 41 {Isolation and characterisation of chitosanase from Bacillus sp . 739 strain}; Aktuganov GE et al.; The specific nature of the chitosanase activity of the strain Bacillus sp . 739 has been determined . Maximum enzyme activity was observed in a medium containing the biomass of the fruiting bodies of the fungus Macrolepiota procera . The chitosanase was purified to homogeneity using chromatography on DEAE-Sephadex A-50 and Toyopearl HW-50 . The molecular weight of the enzyme, assessed by electrophoresis (the Laemmli procedure) approximated 46 kDa . Temperature and pH optima of the purified chitosanase were in the ranges 45-55 degrees C and 6.0-6.5, respectively . Time to half-maximum inactivation of the enzyme at 50 degrees C was equal to 1 h . With colloidal chitosan as the substrate, the value of K(M) of the purified chitosanase was equal to 25 mg/ml . The enzyme also exhibited a weak ability to hydrolyze colloidal chitin. J Infect Dis, 2003 Nov 1, 188(9), 1332 - 5 Epub 2003 Oct 15. Nosocomial Mycobacterium bovis-bacille Calmette-Guérin infections due to contamination of chemotherapeutics: case finding and route of transmission; Vos MC et al.; We studied nosocomial infections due to Mycobacterium bovis bacille Calmette-Guerin (BCG) Onco-TICE bacteria, transmitted by contamination of medication prepared in BCG Onco-TICE-contaminated hoods in the pharmacy, in 5 immunocompromised patients at 3 hospitals . The BCG strains cultured from the patients had the same DNA profile as the BCG Onco-TICE strain used for bladder instillation . To prevent these infections, a change from open to closed preparation was made; strictly separated preparation in time of BCG Onco-TICE instillation and chemotherapy was enforced, the biological safety cabinet was disinfected between preparations, and gloves were changed between preparations. Mycorrhiza, 2003 Oct, 13(5), 249 - 56 Epub 2003 Feb 15. Influence of a Bacillus sp . on physiological activities of two arbuscular mycorrhizal fungi and on plant responses to PEG-induced drought stress; Vivas A et al.; The effects of bacterial inoculation (Bacillus sp.) on the development and physiology of the symbiosis between lettuce and the arbuscular mycorrhizal (AM) fungi Glomus mosseae (Nicol . and Gerd.) Gerd . and Trappe and Glomus intraradices (Schenck and Smith) were investigated . Plant growth, mineral nutrition and gas-exchange values in response to bacterial inoculation after PEG-induced drought stress were also evaluated . In AM plants, inoculation with Bacillus sp . enhanced fungal development and metabolism, measured as succinate dehydrogenase (SDH) and alkaline phosphatase (ALP) activities, more than plant growth . Under non-stressed conditions, G . intraradices colonization increased all plant physiological values to a higher extent when in dual inoculation with the bacterium . Under stress conditions, the bacterium had an important stimulatory effect on G . intraradices development . Under such conditions, the effects of the bacterium on photosynthetic rate, water use efficiency (WUE) and stomatal conductance of lettuce plants differed with the fungus species . Plant-gas exchange was enhanced in G . intraradices- and reduced in G . mosseae-colonized plants when co-inoculated with Bacillus sp . Thus, the effects of each fungus on plant physiology were modulated by the bacterium . Stress was detrimental, particularly in G . intraradices-colonized plants without the bacterium, reducing intra and extraradical mycelium growth and vitality (SDH), as well as plant-gas exchange . Nevertheless, Bacillus sp . inoculation improved all these plant and fungal parameters to the same level as in non-stressed plants . The highest amount of alive and active AM mycelium for both fungi was obtained after co-inoculation with Bacillus sp . These results suggest that selected free-living bacteria and AM fungi should be co-inoculated to optimize the formation and functioning of the AM symbiosis in both normal and adverse environments. Appl Microbiol Biotechnol, 2004 May, 64(4), 497 - 504 Epub 2003 Oct 31. Diglucosyl-glycerolipids from the marine sponge-associated Bacillus pumilus strain AAS3: their production, enzymatic modification and properties; Ramm W et al.; The marine strain Bacillus pumilus strain AAS3, isolated from the Mediterranean sponge Acanthella acuta, produced a diglucosyl-glycerolipid, 1,2-O-diacyl-3-{beta-glucopyranosyl-(1-6)-beta-glucopyranosyl)}glycerol, with 14-methylhexadecanoic acid and 12-methyltetradecanoic acid as the main fatty acid moieties (GGL11) . On a 30 l scale, using artificial seawater supplemented with glucose (20 g/l), yeast extract (10 g/l), and suitable nitrogen/phosphate sources, growth-associated glycoglycerolipid production reached its maximum yield of 90 mg/l after 11 h . Lipase-catalyzed modification of the native substance led to the deacylated parent compound (GG11), which could be reacylated using the same enzyme system to afford a new dipentenoyl-diglucosylglycerol (GGL12) as the major product upon addition of 4-pentenoic acid to the medium . GGL11 decreased the surface tension of water from 72 mN/m to 29 mN/m and the interfacial tension of the water/ n-hexadecane system from 44 to 5 mN/m . Anti-tumor-promoting studies on this class of diglucosyl glycerol products showed that the carbohydrate/glycerol backbone (GG11) has a more potent inhibitory activity than the acylated compounds . The diglucosyl-glycerol GG11 strongly inhibited growth of the tumor cell lines HM02 and Hep G2 (50% inhibition at approximately 1 microg/ml), while the glycerolipids GGL11 and GGL12 were less active or had no effect . Appl Microbiol Biotechnol, 2004 Apr, 64(2), 243 - 9 Epub 2003 Oct 31. Identification and functional characterization of a type I signal peptidase gene of Bacillus megaterium DSM319; Nahrstedt H et al.; The sipM gene of Bacillus megaterium encoding a type I signal peptidase (SPase) was isolated and structurally characterized . RNA analysis revealed a transcript size in accordance with a bicistronic operon comprising sipM and an adjacent open reading frame . Inactivation of sipM by targeted gene disruption could not be achieved, indicating its essential role for cell viability since there might be no other type I SPase of major importance present in B . megaterium . Plasmid-assisted amplification of the gene resulted in an increase in activity of the heterologous glucanase used as an extracellular reporter, suggesting a potential bottleneck for protein secretion within this species. Appl Microbiol Biotechnol, 2004 Apr, 64(2), 187 - 91 Epub 2003 Oct 31. Artificial carrier for oxygen supply in biological systems; Dey ES et al.; Several poly (dimethylsiloxanes) (PDMS) copolymers of dimethylsiloxane (DMS) with ethylene or propylene oxide were tested as artificial carriers for the delivery of oxygen to biological systems . Copolymers with a DMS content of 33% or lower enhanced glucose oxidation by 200% in contrast to the 25% increase produced by the same concentration of perfluorodecalin . When 0.05% of the copolymer with 18% DMS was included in the growth media of Bacillus thuriginensis, the biomass (growth rate) increased 1.5-fold . With 0.1% of this copolymer, actinorhodin production by Streptomyces coelicolor A3 (2) occurred in half the normal time and with an increased yield . In conclusion, these PDMS copolymers are a good alternative to perfluorodecalin as oxygen carriers in biotechnological processes. FEMS Microbiol Lett, 2003 Oct 24, 227(2), 189 - 95 Mosquito larvicidal activity of transgenic Anabaena PCC 7120 expressing toxin genes from Bacillus thuringiensis subsp . israelensis; Khasdan V et al.; Genes encoding the mosquito larvicidal toxins Cry4Aa, Cry11Aa, Cyt1Aa and the regulatory P20 from Bacillus thuringiensis subsp . israelensis were introduced into the nitrogen-fixing, filamentous cyanobacterium Anabaena PCC 7120 for expression under control of two strong promoters P(psbA) and P(A1) . The clone pRVE4-ADRC displayed toxicity against fourth-instar larvae of Aedes aegypti, the highest ever achieved in cyanobacteria . It was about 2.5-fold more toxic than the respective clone without cyt1Aa {Wu et al., Appl . Environ . Microbiol . 63 (1997) 4971-4975} . Cyt1Aa synergized the combination of Crys by about five-fold . Consistently, the lethal times exerted by pRVE4-ADRC were also reduced (it killed exposed larvae more quickly) . This clone may become a useful biological control agent which reduces the probability of resistance development in the target organisms {Wirth et al., Proc . Natl . Acad . Sci . USA 94 (1997) 10536-10540}. Biochem Biophys Res Commun, 2003 Nov 14, 311(2), 386 - 90 Real-time molecular beacon NASBA reveals hblC expression from Bacillus spp . in milk; Gore HM et al.; Nucleic acid sequence-based amplification (NASBA) was applied in combination with a fluorescein-conjugated molecular beacon specific for a sequence flanked by transcript-specific primers in order to monitor hblC enterotoxin gene expression in real-time from milk separately contaminated with Bacillus amyloliquefaciens, Bacillus cereus, and Bacillus circulans . Maximal enterotoxin expression was noted following 16, 15, and 16 h, respectively, when grown in artificially contaminated nonfat dried milk incubated aerobically at 32 degrees C, corresponding to 1.6 x 10(5), 5 x 10(7), and 9.8 x 10(4)cfu/ml, for B . amyloliquefaciens, B . cereus, and B . circulans, respectively . This RNA amplification assay allows for simultaneous detection and confirmation of target transcripts in a closed tube format and may be performed in a high DNA background . The development of a rapid, sensitive, real-time method to quantitate the expression of virulence genes in pathogenic spore-formers is useful in shelf life determination of foods and other quality assurance measures. Biosci Biotechnol Biochem, 2003 Oct, 67(10), 2304 - 6 Increased production of antioxidative sesaminol glucosides from sesame oil cake through fermentation by Bacillus circulans strain YUS-2; Ohtsuki T et al.; Bacillus circulans strain YUS-2 was isolated as the strongest antioxidant-producer in fermentation of sesame oil cake (SOC, defatted residue yielded from sesame seed oil production) . Two major strong antioxidants from fermented SOC were purified and identified as known sesaminol triglucoside and sesaminol diglucoside, however, our results demonstrated that the fermentation process with B . circulans YUS-2 was highly effective to gain the extraction efficiency of the sesaminol glucosides. Biosci Biotechnol Biochem, 2003 Oct, 67(10), 2132 - 8 Endophytic colonization of balloon flower by antifungal strain Bacillus sp . CY22; Cho SJ et al.; Endophytic Bacillus sp . CY22 was previously isolated from the root interior of the balloon flower (Platycodon grandiflorum) (Cho et al., Biosci . Biotechnol . Biochem., 66, 1270-1275 (2002)) . Three-month-old balloon flower seedlings were inoculated with 10(7) cfu/ml of strain CY22R3, a rifampicin-resistant strain of CY22, and external and internal root colonization was assessed 2 and 4 weeks later . After inoculation, large numbers of bacteria were observed on the root surface by scanning electron microscopy . More detailed studies using optical and transmission electron microscopy confirmed that Bacillus sp . CY22 was endophytically established within intercellular spaces, cortical cells, and aerenchymas of root . Also, Bacillus sp . CY22 showed antibiotic activities against several phytopathogens by producing the antibiotic iturin A . In the pot test, root rot of balloon flower seedlings caused by Rhizoctonia solani was suppressed when the Bacillus sp . CY22R3 was inoculated into the soil. Vaccine, 2003 Dec 1, 21(32), 4722 - 7 Role of L-lysine HCl in immunopotentiation towards development of suitable tuberculosis vaccination; Dasgupta S et al.; L-Lysine HCl is being proposed to be a possible biocompatible adjuvant to enhance immune response by virtue of its probable non-specific bridging action and cellular proliferation properties . This proposal has been tried to be substantiated by carrying out experimentation where L-lysine HCl has been used as an adjuvant (various groups based on mode of application and frequency of booster dose were designed) in tuberculosis vaccination experiments with heat killed Mycobacterium tuberculosis (MTB) and Bacille Calmette Guerin (BCG) . Antibody titre has been followed in all the experiments as a measure of immune response . Amongst the various groups designed, group 1A (L-lysine HCl was given at a separate site as that of the antigen; lysine booster was given to this group intermittently, i.e . lysine given on 0th, 7th, 14th, 21st days of immunization) came out as the stand-alone leader . This mode and frequency of application was then compared with a group which received a standard adjuvant, viz . alhydrogel . Results were obtained which showed the following order in terms of decreasing antibody titre: alhydrogel group > lysine group > control group . Considering the biocompatible nature of lysine in comparison with the reportedly hazardous nature of alum adjuvants, we propose L-lysine HCl as a probable adjuvant in vaccination. J Am Chem Soc, 2003 Nov 5, 125(44), 13442 - 50 Regio- and enantioselective alkane hydroxylation with engineered cytochromes P450 BM-3; Peters MW et al.; Cytochrome P450 BM-3 from Bacillus megaterium was engineered using a combination of directed evolution and site-directed mutagenesis to hydroxylate linear alkanes regio- and enantioselectively using atmospheric dioxygen as an oxidant . BM-3 variant 9-10A-A328V hydroxylates octane at the 2-position to form S-2-octanol (40% ee) . Another variant, 1-12G, also hydroxylates alkanes larger than hexane primarily at the 2-position but forms R-2-alcohols (40-55% ee) . These biocatalysts are highly active (rates up to 400 min(-1)) and support thousands of product turnovers . The regio- and enantioselectivities are retained in whole-cell biotransformations with Escherichia coli, where the engineered P450s can be expressed at high levels and the cofactor is supplied endogenously. J Agric Food Chem, 2003 Nov 5, 51(23), 6823 - 7 Rapid digestion of Cry34Ab1 and Cry35Ab1 in simulated gastric fluid; Herman RA et al.; Two genes were identified in Bacillus thuringiensis Berliner (Bt) that code for the proteins that comprise a Cry34Ab1/Cry35Ab1 binary insecticidal crystal protein . Maize, Zea mays L., plants have been transformed to express the Cry34Ab1/Cry35Ab1 proteins, and as a result, these plants are resistant to attack by western corn rootworm, Diabrotica virgifera virgifera LeConte, a major pest in the Midwestern corn-growing area of the U.S.A . As part of the safety assessment for the proteins, digestibility studies were conducted . Digestion experiments with both proteins demonstrated rapid degradation in simulated gastric fluid, comparable to other registered plant-incorporated protectants . Quantitative and qualitative approaches for determining digestibility are illustrated. Biochim Biophys Acta, 2003 Nov 3, 1652(1), 52 - 63 A proposed mechanism for the thermal denaturation of a recombinant Bacillus halmapalus alpha-amylase--the effect of calcium ions; Nielsen AD et al.; The thermal stability of a recombinant alpha-amylase from Bacillus halmapalus alpha-amylase (BHA) has been investigated using circular dichroism spectroscopy (CD) and differential scanning calorimetry (DSC) . This alpha-amylase is homologous to other Bacillus alpha-amylases where crystallographic studies have identified the existence of three calcium binding sites in the structure . Denaturation of BHA is irreversible with a T(m) of approximately 89 degrees C and DSC thermograms can be described using a one-step irreversible model . A 5 degrees C increase in T(m) in the presence of 10-fold excess CaCl(2) was observed . However, a concomitant increase in the tendency to aggregate was also observed . The presence of 30-40-fold excess calcium chelator (ethylenediaminetetraacetic acid (EDTA) or ethylene glycol-bis{beta-aminoethyl ether} N,N,N',N'-tetraacetic acid (EGTA)) results in a large destabilization of BHA, corresponding to about 40 degrees C lower T(m) as determined by both CD and DSC . Ten-fold excess EGTA reveals complex DSC thermograms corresponding to both reversible and irreversible transitions, which probably originate from different populations of BHA/calcium complexes . Combined interpretation of these observations and structural information on homologous alpha-amylases forms the basis for a suggested mechanism underlying the inactivation mechanism of BHA . The mechanism includes irreversible thermal denaturation of different BHA/calcium complexes and the calcium binding equilibria . Furthermore, the model accounts for a temperature-induced reversible structural change associated with calcium binding. J Biotechnol, 2003 Nov 6, 105(3), 255 - 60 Magnetic alginate microparticles for purification of alpha-amylases; Safarikova M et al.; Spherical magnetic alginate microparticles (25-60 microm in diameter) were prepared using the microemulsion system, with water-saturated 1-pentanol as the organic phase . The limited solubility of 1-pentanol in water enabled simple removal of the organic solvent from the prepared beads with water solution . The prepared alginate microparticles were used as magnetic affinity adsorbents for specific purification of alpha-amylases . Enzyme activity was eluted by 1.0 M maltose . alpha-Amylases from Bacillus amyloliquefaciens and porcine pancreatic acetone powder were purified 9- and 12-fold with 88 and 96% activity recovery, respectively. J Transl Med . 2003 Oct 10;1(1):7. Interferon-gamma Added During Bacillus Calmette-Guerin Induced Dendritic Cell Maturation Stimulates Potent Th1 Immune Responses; Shankar G et al.; Dendritic cells (DC) are increasingly prepared in vitro for use in immunotherapy trials . Mature DC express high levels of surface molecules needed for T cell activation and are superior at antigen-presentation than immature DC . Bacillus Calmette-Guerin (BCG) is one of several products known to induce DC maturation, and interferon (IFN)-gamma has been shown to enhance the activity of DC stimulated with certain maturation factors . In this study, we investigated the use of IFN-gamma in combination with the powerful maturation agent, BCG . The treatment of immature DC with IFN-gamma plus BCG led to the upregulation of CD54, CD80, and CD86 in comparison with BCG treatment alone . In MLR or recall immune responses, the addition of IFN-gamma at the time of BCG-treatment did not increase the number of antigen-specific T cells but enhanced the development of IFN-gamma-producing Th1 cells . In primary immune responses, on the other hand, BCG and IFN-gamma co-treated DC stimulated higher proportions of specific T cells as well as IFN-gamma secretion by these T cells . Thus the use of IFN-gamma during BCG-induced DC maturation differentially affects the nature of recall versus naive antigen-specific T-cell responses . IFN-gamma co-treatment with BCG was found to induce IL-12 and, in some instances, inhibit IL-10 secretion by DC . These findings greatly enhance the potential of BCG-matured dendritic cells for use in cancer immunotherapy. Proteins, 2003 Nov 15, 53(3), 597 - 606 Unfolding of the cold shock protein studied with biased molecular dynamics; Morra G et al.; The cold shock protein from Bacillus caldolyticus is a small beta-barrel protein that folds in a two-state mechanism . For the native protein and for several mutants, a wealth of experimental data are available on stability and folding, so that it is an optimal system to study this process . We compare data from unfolding simulations (trajectories of 5 and up to 12 ns) obtained with a bias potential at room temperature and from unbiased thermal unfolding simulations with experimental data . The unfolding patterns derived from the trajectories starting from different native-like conformations and subject to different unfolding conditions agree . The transition state found in the simulations of unfolding is close to the native structure in agreement with experiment . Moreover, a lower value of the free energy barrier of unfolding was found for the mutant R3E than for the mutant E46A and the native protein, as indicated by experimental data . The first unfolding event involves the three-stranded beta-sheet whose decomposition corresponds to the transition state . In contrast to conclusions drawn from experiments, we found that the two-stranded beta-strand forms the most stable substructure, which decomposes very late in the unfolding process . However, assuming that this structure forms very early in the folding process, our findings would not contradict the experiments but require a different interpretation of them . Mol Pharmacol, 2003 Nov, 64(5), 1189 - 98 Ceramide is involved in r(+)-methanandamide-induced cyclooxygenase-2 expression in human neuroglioma cells; Ramer R et al.; Cannabinoids have recently been shown to induce the expression of the cyclooxygenase-2 (COX-2) isoenzyme in H4 human neuroglioma cells . Using this cell line, the present study investigates the contribution of the second messenger ceramide to this signaling pathway . Incubation of cells with the endocannabinoid analog R(+)-methanandamide (R(+)-MA) was associated with an increase of intracellular ceramide levels . Enhancement of ceramide formation by R(+)-MA was abolished by fumonisin B1, a ceramide synthase inhibitor, whereas inhibitors of neutral sphingomyelinase (spiroepoxide, glutathione) and serine palmitoyltransferase (l-cycloserine, ISP-1) were inactive in this respect . R(+)-MA caused a biphasic activation of the p38 and p42/44 mitogen-activated protein kinases (MAPKs), with phosphorylation peaks occurring after 15-min and 4- to 8-h treatments, respectively . Inhibition of ceramide synthesis with fumonisin B1 was associated with a suppression of R(+)-MA-induced delayed phosphorylations of p38 and p42/44 MAPKs and subsequent COX-2 expression . The involvement of ceramide in COX-2 expression was corroborated by findings showing that C2-ceramide and neutral sphingomyelinase from Bacillus cereus caused concentration-dependent increases of COX-2 expression that were suppressed in the presence of 4-(4-fluorophenyl)-2-(4-methylsulfonylphenyl)-5-(4-pyridyl)imidazol (SB203580, a p38 MAPK inhibitor) or 2'-amino-3'-methoxyflavone (PD98059, a p42/44 MAPK activation inhibitor) . In contrast, dihydro-C2-ceramide being used as a negative control did not induce MAPK phosphorylation and COX-2 expression . Collectively, our results demonstrate that R(+)-MA induces COX-2 expression in human neuroglioma cells via synthesis of ceramide and subsequent activation of p38 and p42/44 MAPK pathways . Induction of COX-2 expression via ceramide represents a hitherto unknown mechanism by which cannabinoids mediate biological effects within the central nervous system. Infect Immun, 2003 Nov, 71(11), 6411 - 9 Revaccination of neonatal calves with Mycobacterium bovis BCG reduces the level of protection against bovine tuberculosis induced by a single vaccination; Buddle BM et al.; Cattle may provide a suitable model for testing ways of improving tuberculosis vaccine efficacy in human infants . A vaccination and challenge study was undertaken in calves to determine the optimal time to vaccinate neonatal animals with Mycobacterium bovis bacillus Calmette-Guerin (BCG) for protection against tuberculosis and to determine whether revaccination with BCG was beneficial . Calves (10 per group) were vaccinated with BCG within 8 h of birth or at 6 weeks of age, when immune responses to antigens of environmental mycobacteria were detectable, or vaccinated at birth and revaccinated at 6 weeks . A control group was not vaccinated . BCG vaccination at birth induced strong antigen-specific gamma interferon (IFN-gamma) and interleukin-2 (IL-2) responses and antigen-specific activation in CD4(+), CD8(+), and WC1(+) gammadelta T-cell subsets from blood . The proportions of animals per group with macroscopic tuberculous lesions after challenge were 0/10 for BCG at birth, 1/9 for BCG at 6 weeks, 4/10 for the revaccinated group, and 10/10 for the nonvaccinated group . There was no significant difference in the levels of protection between groups vaccinated at birth or at 6 weeks, while animals vaccinated both at birth and at 6 weeks had significantly less protection than those vaccinated only at birth . The revaccinated calves that subsequently developed tuberculous lesions had significantly stronger IFN-gamma and IL-2 responses to bovine purified protein derivative after the BCG booster than those in the same group that did not develop lesions . The results indicated that BCG vaccination at birth induced a high level of immunity and that the sensitization of very young animals to antigens of environmental mycobacteria by 6 weeks of age did not affect the effectiveness of BCG . However, BCG revaccination of these young animals was contraindicated. Biotechnol Lett, 2003 Sep, 25(18), 1555 - 62 Sequence analysis of cyclodextrin glycosyltransferase from the alkaliphilic Bacillus agaradhaerens; Martins RF et al.; The gene encoding an alkaline active cyclodextrin glycosyltransferase (CGTase) from the alkaliphilic B . agaradhaerens LS-3C was cloned and sequenced . It encodes a mature polypeptide of 679 amino acids with a molecular mass of 76,488 Da . The deduced amino acid sequence of the mature CGTase revealed 99 and 95% identity to the CGTase sequences from the other B . agaradhaerens strains, DSM 8721T and 9948, respectively . The next closest identity was of 59% with B . clarkii enzyme . CGTases from B . agaradhaerens, B . clarkii, and B . firmus/lentus formed a phylogenetically separated cluster from the other CGTases of Bacillus spp . origin . A number of usually conserved residues in the CGTases were found to be replaced in the sequence of B . agaradhaerens enzyme . The sequence analysis indicated the enzyme to be close to the so-called 'intermediary enzymes' in the alpha-amylase family. Biotechnol Lett, 2003 Sep, 25(18), 1537 - 43 Integrated immobilized cell reactor-adsorption system for beta-cyclodextrin production: a model study using PVA-cryogel entrapped Bacillus agaradhaerens cells; Martins RF et al.; Production of cyclodextrins (CDs) by immobilized cells of the alkaliphilic Bacillus agaradhaerens LS-3C with integrated product recovery was studied . The microorganism was entrapped in polyvinyl alcohol-cryogel beads and used as a convenient source of immobilized cyclodextrin glycosyltransferase (CGTase) . On activation by incubation in the cultivation medium containing 1% (w/v) starch, the entrapped cells multiplied and secreted CGTase with an activity of 2-3 mg beta-cyclodextrin h(-1) g(-1) beads . The immobilized biocatalyst exhibited maximum activity at pH 9 and 50 degrees C, and formed cyclodextrins comprising 92-94% beta-CD and remaining alpha-CD . The cyclodextrin product from the immobilized cell bioreactor was continuously recovered by adsorption to Amberlite XAD-4 in a recycle batch mode . The product adsorption was facilitated at low temperature while hot water was used for elution. Arch Mal Coeur Vaiss, 2003 Sep, 96(9), 923 - 6 {Cardiobacterium hominis endocarditis . A case report}; Balcou-Leroy E et al.; We report the case of a Cardiobacterium hominis endocarditis causing an acute mitral insufficiency complicated of left heart failure . The patient has been treated after a few days by surgical valvuloplasty . Cardiobacterium hominis is a bacteria of the HACCEK group, bacille gram-negative, sometimes anaerobic, difficult to isolate . Recently, Polymerase Chain Reaction analysis appears to be effective for the the diagnosis in the identification of fastidious micro-organisms like Cardiobacterium hominis . We have reviewed in the literature 71 cases of Cardiobacterium hominis endocarditis; clinical presentation is often sub-acute, the bacteriological diagnosis is based on hemocultures for which the culture is slow and require enriched environments . Hemodynamic and thrombo-embolic complications are frequent because of the high pathogenicity of the bacteria which provides big and friable vegetations . Despite a high sensibility to antibiotherapy, surgical intervention is often required. AIDS, 2003 Nov 7, 17(16), 2351 - 5 Immunogenicity of an inactivated mycobacterial vaccine for the prevention of HIV-associated tuberculosis: a randomized, controlled trial; Vuola JM et al.; OBJECTIVE: Prior to the widespread use of Mycobacterium bovis, Bacille Calmette-Guerin (BCG), inactivated whole cell mycobacterial vaccines had been shown effective in the prevention of tuberculosis . The present study was conducted to determine the safety and immunogenicity of an inactivated whole cell mycobacterial vaccine in persons with HIV infection.DESIGN Randomized, controlled trial . METHODS: A total of 39 HIV-positive patients with prior BCG immunization and CD4 cell counts >/= 200 x 10(6) cells/l were randomized to five doses of inactivated Mycobacterium vaccae (MV) vaccine or control vaccine (CV) . Lymphocyte proliferation (LPA) and interferon gamma (IFN-gamma) responses to mycobacterial antigens were assayed at baseline, after three and five doses of vaccine and > 1 year later . Parallel studies were conducted in 10 HIV-negative subjects with prior BCG immunization . RESULTS: Among HIV-positive patients, 19 MV recipients had higher LPA and IFN-gamma responses to MV sonicate than 20 CV recipients after three and five doses of vaccine and > 1 year later . LPA responses to Mycobacterium tuberculosis whole cell lysate increased over time in both groups consistent with prior BCG immunization and current antiretroviral therapy; after three doses, responses were boosted to higher levels in MV subjects than CV subjects . LPA responses to WCL were also boosted in HIV-negative MV recipients . Immunization was safe and had no adverse effects on HIV viral load or CD4 cell count . CONCLUSIONS: In BCG-primed, HIV-positive and HIV-negative subjects, MV induces durable cellular immune responses to a new mycobacterial antigen and boosts pre-existing responses to WCL . MV is a candidate for clinical trials for the prevention of HIV-associated tuberculosis. Curr Microbiol, 2003 Sep, 47(3), 226 - 30 Toxicity of Bacillus sphaericus LP1-G against susceptible and resistant Culex quinquefasciatus and the cloning of the mosquitocidal toxin gene; Shi YX et al.; Bacillus sphaericus LP1-G, belonging to flagellar serotype H3, has been found to have moderate toxicity against two resistant Culex quinquefasciatus colonies (RLCq1 and RLCq2) and the susceptible contrast (SLCq) . With an aim of screening mosquitocidal acting factor, a partial genome library was prepared from a partial HindIII digest of the total DNA from Bacillus sphaericus LP1-G . Two thousand twenty Escherichia coli clones were screened for toxicity against susceptible SLCq, and a toxic clone, designated E-UL68, was chosen for further study . The recombinant E-UL68 performed toxicity against both susceptible and two resistant colonies, having the same level of toxicity as that of wide-type strain LP1-G . Sequence analysis revealed that the inserted fragment was composed of 3876 nucleotides and contained a complete gene, whose sequence was identical to that of the mtx gene from B . sphaericus SSII-1 . Because the binary toxin produced during sporulation of strain LP1-G has no activity against the target mosquitoes, this indicates that the Mtx toxin or other active factors might perhaps be responsible for the toxicity of LP1-G against different colonies of mosquito larvae. Am J Clin Pathol, 2002 Dec, 118 Suppl, S3 - 17 Tuberculosis . Advances in laboratory diagnosis and drug susceptibility testing; Eichbaum Q et al.; Laboratory diagnosis of mycobacterial infection is complicated by the fastidious growth requirements of the bacillus organism . Delays in diagnosis can impede effective treatment and surveillance of the disease . Control of Mycobacterium tuberculosis has also been aggravated by the emergence of multidrug-resistant tuberculosis, and enhanced methods of antibiotic susceptibility testing are needed . Molecular laboratory techniques and advances in rapid culture methods have led to the development of diagnostic and drug susceptibility tests that are rapid and reliable . This article describes recent innovations in the molecular diagnosis and antibiotic susceptibility testing of M tuberculosis. J Immunol, 2003 Nov 1, 171(9), 4689 - 99 CD4+ T cells mediate IFN-gamma-independent control of Mycobacterium tuberculosis infection both in vitro and in vivo; Cowley SC et al.; Although IFN-gamma is necessary for survival of Mycobacterium tuberculosis infection in people and animal models, it may not be sufficient to clear the infection, and IFN-gamma is not a reliable correlate of protection . To determine whether IFN-gamma-independent mechanisms of immunity exist, we developed a murine ex vivo culture system that directly evaluates the ability of splenic or lung lymphocytes to control the growth of M . tuberculosis within infected macrophages, and that models in vivo immunity to tuberculosis . Surprisingly, CD4(+) T cells controlled >90% of intracellular M . tuberculosis growth in the complete absence of IFN-gamma stimulation of macrophages, via a NO-dependent mechanism . Furthermore, bacillus Calmette-Guerin-vaccinated IFN-gamma-deficient mice exhibited significant protection against M . tuberculosis challenge that was lost upon depletion of CD4(+) T cells . These findings demonstrate that CD4(+) T cells possess IFN-gamma-independent mechanisms that can limit the growth of an intracellular pathogen and are dominant in secondary responses to M . tuberculosis. J Mol Biol, 2003 Oct 31, 333(4), 721 - 45 Optimal determination of particle orientation, absolute hand, and contrast loss in single-particle electron cryomicroscopy; Rosenthal PB et al.; A computational procedure is described for assigning the absolute hand of the structure of a protein or assembly determined by single-particle electron microscopy . The procedure requires a pair of micrographs of the same particle field recorded at two tilt angles of a single tilt-axis specimen holder together with the three-dimensional map whose hand is being determined . For orientations determined from particles on one micrograph using the map, the agreement (average phase residual) between particle images on the second micrograph and map projections is determined for all possible choices of tilt angle and axis . Whether the agreement is better at the known tilt angle and axis of the microscope or its inverse indicates whether the map is of correct or incorrect hand . An increased discrimination of correct from incorrect hand (free hand difference), as well as accurate identification of the known values for the tilt angle and axis, can be used as targets for rapidly optimizing the search or refinement procedures used to determine particle orientations . Optimized refinement reduces the tendency for the model to match noise in a single image, thus improving the accuracy of the orientation determination and therefore the quality of the resulting map . The hand determination and refinement optimization procedure is applied to image pairs of the dihydrolipoyl acetyltransferase (E2) catalytic core of the pyruvate dehydrogenase complex from Bacillus stearothermophilus taken by low-dose electron cryomicroscopy . Structure factor amplitudes of a three-dimensional map of the E2 catalytic core obtained by averaging untilted images of 3667 icosahedral particles are compared to a scattering reference using a Guinier plot . A noise-dependent structure factor weight is derived and used in conjunction with a temperature factor (B=-1000A(2)) to restore high-resolution contrast without amplifying noise and to visualize molecular features to 8.7A resolution, according to a new objective criterion for resolution assessment proposed here. J Mol Biol, 2003 Oct 31, 333(4), 711 - 20 Solution structural study of BlaI: implications for the repression of genes involved in beta-lactam antibiotic resistance; Melckebeke HV et al.; beta-Lactamase and penicillin-binding protein PBP2' mediate staphylococcal resistance to beta-lactam antibiotics, which are otherwise highly clinically effective . Two repressors (BlaI and MecI) regulate expression of these inducible proteins . Here, we present the first solution structure of the 82 amino acid residue DNA-binding domain of Bacillus licheniformis BlaI which is very similar in primary sequence to the medically significant Staphyloccocal BlaI and MecI proteins . This structure is composed of a compact core of three alpha-helices and a three-stranded beta-sheet typical of the winged helix protein (WHP) family . The protein/DNA complex was studied by NMR chemical shift comparison between the free and complexed forms of BlaI . Residues involved in DNA interaction were identified and a WHP canonical model of interaction with the operators is proposed . In this model, specific contacts occur between the base-pairs of the TACA motif and conserved amino acid residues of the repressor helix H3 . These results help toward understanding the repression and induction mechanism of the genes coding for beta-lactamase and PBP2'. Med Parazitol (Mosk), 2003 Jul-Sep, (3), 18 - 20 {Indication of Borrelia burgdorferi sensu lato in the taiga tick larvae obtained from spontaneously infected females}; Nefedova VV; Hungry larvae from 17 clutches by spontaneously infected Ixodes persulcatus females were examined by inoculation of the BSK II medium, by microscopy of fixed smears, and by polymerase chain reaction (PCR) with the primers specific toward the conserved sequences of Borrelia burgdorferi 16S ribosomal RNA gene . A study of 781 larvae individually or as pools could not reveal Borrelia DNA by PCR . Inoculations of the BSK II medium with 600 larvae yielded no positive results either; immobile spiral forms of Bacillus spp . were detected in 16 (26.7 +/- 5.7%) cultures of the larvae obtained from 7 females . Microscopy of 1416 fixed smears showed typical Borrelia in 7 (0.5 +/- 0.4%) specimens of larvae resulting from the same hatch; spirochaete-like cells were present in 13 (0.9 +/- 0.5%) cases . The single female from those of the 17 hatches transmitted Borrelia through eggs to filial-generation larvae . The infection rate of the larvae emerged from the eggs of this hatch was 7% and their individual infection rates were 0.4-0.8 Borrelia per 100 fields of vision . The findings suggest that there may a transovarian transmission of Borrelia in I . persulcatus ticks, but its likelihood is very little. J Ind Microbiol Biotechnol, 2003 Oct, 30(10), 606 - 12 Epub 2003 Oct 16. Solubilization of cobalt from ocean nodules at neutral pH-a novel bioprocess; Mukherjee A et al.; A marine organism ( Bacillus M1) isolated from Indian Ocean manganese nodules was characterized . The organism grew well in artificial seawater medium, at near neutral pH, 30 degrees C and 0.25 M NaCl, and showed MnO(2)-reducing activity . Growing cultures of Bacillus M1 as well as cell-free spent liquor from fully-grown cultures were employed to extract metals from the nodules . The spent liquor of cultures of the organism could dissolve around 45% cobalt (Co) at a pH of 8.2 in 2 h . Co recovery by this treatment was comparable to that in acidic leaching with 2.5 M hydrochloric acid solutions, and was independent of pulp density (w/v ratio) . The amount of Co dissolved was beyond the thermodynamic solubility limit in aqueous solution at a pH of 8.2 . It is inferred that the metabolites present in the spent liquor played a pivotal role in complexing the Fe (III) phase, solubilizing Co in the process . Partial characterization of spent liquor by spot tests, UV visible spectroscopy and FTIR spectroscopy, showed the presence of siderophore-like phenolic compound(s) with an attached carboxyl group that might form soluble organic complexes with Fe (III). Mol Immunol, 2003 Dec, 40(8), 555 - 62 Major linear IgE epitopes of mountain cedar pollen allergen Jun a 1 map to the pectate lyase catalytic site; Midoro-Horiuti T et al.; Resolution of the 3D structures and IgE epitopes of allergens may identify common or conserved features of allergens . Jun a 1, the predominant allergen in mountain cedar pollen, was chosen as a model for identifying common structural and functional features among a group of plant allergens . In this study, synthetic, overlapping peptides of Jun a 1 and sera from patients allergic to mountain cedar pollen were used to identify linear epitopes . A 3D model of Jun a 1 was produced using the Bacillus subtiles pectate lyase (PL) as a template and validated with biophysical measurements . This allowed mappings of four IgE binding sites on Jun a 1 . Two of the epitopes mapped to turns or loops on the surface of the model structure . The other two epitopes mapped to the beta-sheet region, homologous to the catalytic site of PL . This region of Jun a 1 is highly conserved in the group 1 allergens from other cedar trees as well as microbial PLs . The finding that two out of three major IgE epitopes map to highly conserved catalytic regions of group 1 cedar allergens may help to explain the high degree of cross-reactivity between cedar pollen allergens and might represent a pattern of reactivity common to other allergens with catalytic activity. J Antibiot (Tokyo), 2003 Aug, 56(8), 716 - 20 SNA-60-367 components, new peptide enzyme inhibitors of aromatase: structure of the fatty acid side chain and amino acid sequence by mass spectrometry; Esumi Y et al.; SNA-60-367 components, new peptide enzyme inhibitors of aromatase, were isolated from the culture broth of soil bacterium, Bacillus sp . SNA-60-367 . These inhibitors are a family of acylated decapeptides that differ from each other in terms of amino acid composition and the nature of the fatty acid side chain . The structures of the fatty acid moieties were shown to be (3-hydroxy)heptadecanoic acid and (3-hydroxy)hexadecanoic acid that possess normal-, iso- or anteiso-type alkyl groups . The amino acid sequence of the open form of the lactone ring of the acylpeptides is RCO-L-Glu-D-Orn-L(or D)-Tyr3-D-allo-Thr-L-Glu-D-X1 (Ala, Aba or Val)-L-Pro-L-Gln-D(or L)-Tyr-L-X2(10)(Ile or Val)-OH . The lactone ring of SNA-60-367 components is formed between Tyr3 and X2(10). Hum Pathol, 2003 Sep, 34(9), 929 - 38 Pseudomonas pneumonia in infants: an autopsy study; Bonifacio SL et al.; Pseudomonas pneumonia is an uncommon but serious infection in infants, occurring mainly in infants of low birth weight . In this retrospective clinicopathologic correlation study, we reviewed the clinical records and analyzed postmortem lung pathology in 8 infants with pneumonia due to P . aeruginosa . From the histopathology, 2 different pneumonic patterns emerged: a distinctive paucicellular coagulative confluent bronchopneumonia with perivascular bacillary infiltration (7 cases) and a more usual cellular pneumonia without evidence of perivascular organisms (1 case) . Clinically, infants with the first type could be considered immunocompromised and had a precipitous course characterized by signs of sepsis, whereas the infant with the second type (who likely had a more normal immune system) had a relatively protracted course with respiratory failure . We conclude that (1) the pattern of pneumonic inflammation correlates with the immune state of infants, similar to what has been reported in adults; (2) among immunocompromised infants, histopathologic signs of bacteremia are prevalent; and (3) the clinical signs do not correlate with the severity of the pathology at autopsy and may reflect sepsis rather than pneumonia . We speculate that the histopathology in this population reflects the virulence of the organism, as well as the immune status of the host. Protein Pept Lett, 2003 Oct, 10(5), 521 - 4 Purification, crystallization and preliminary X-ray studies of a p-nitrophenylphosphatase from Bacillus stearothermophilus; Ji CN et al.; Thermostable p-nitrophenylphosphatase from Bacillus stearothermophilus has been expressed in Escherichia coli, purified and crystallized . The crystals belong to space group C(2), with unit-cell parameters a = 67.17 A, b = 57.84 A, c = 62.49 A and alpha = 90.0 degrees, beta = 95.4 degrees, gamma = 90.0 degrees . Diffraction data were collected to 1.40 A resolution with a completeness of 94.7% (96.6% for the last shell), an R(fac) value of 0.074 (0.341) and an I/sigma (I) value of 30.1 (2.67). J Theor Biol, 2003 Nov 7, 225(1), 91 - 7 Growth dynamics of Bacillus circulans colony; Komoto A et al.; We have investigated the growth dynamics of Bacillus circulans colony exhibiting the knotted-branching pattern by swarming on a hard agar medium . The knotted-branching pattern consists of many circular clusters, so-called subcolonies, and their trajectories . We analysed the processes of a subcolony because they are presumably the key elements for the formation of knotted-branching pattern . It was found that a subcolony has three processes, i.e . "generation", "growth", and "migration" by microscopic and time-resolved observations . An embryonic small subcolony (child subcolony) formed around an existing subcolony (parent subcolony) grows larger and migrates away from the parent subcolony . We proposed a simple model to explain the migration and the growth processes . It is assumed that the internal part of the subcolony is unfavorable for the bacteria and that the motion of the child subcolony on the agar medium can be modeled using a frictional force . The experimental data were quantitatively analysed in order to compare with models . Our models are consistent with the experimental results on following three points: (1) the radius of a subcolony increases linearly with the incubation time, (2) a subcolony stops just after formation and then starts to migrate suddenly, and (3) the trajectory of a subcolony predicted by the model agrees with the experimental one. Clin Infect Dis, 2003 Nov 1, 37(9), 1226 - 33 Epub 2003 Oct 06. Danish bacille Calmette-Guérin vaccine-induced disease in human immunodeficiency virus-infected children; Hesseling AC et al.; An analysis of isolates of Mycobacterium tuberculosis complex was performed to determine the prevalence of bacille Calmette-Guerin (BCG) disease among human immunodeficiency virus (HIV)-infected children . Speciation was done with polymerase chain reaction; 183 isolates from mycobacterial cultures for 49 HIV-infected patients were analyzed . The Danish Mycobacterium bovis BCG strain was isolated from 5 patients . No cases of Tokyo M . bovis BCG strain disease were detected . All patients were asymptomatic at birth, <12 months of age, and severely immunodeficient at presentation . Four patients had regional axillary adenitis ipsilateral to the vaccination site, and 2 had pulmonary BCG disease . Two patients with regional BCG disease had simultaneous pulmonary M . tuberculosis infection . Although chest radiographic features were similar to those seen in patients with tuberculosis, BCG disease should be considered in HIV-infected infants with right axillary adenitis ipsilateral to the vaccination site . Young, symptomatic, HIV-infected infants are at risk for BCG-related complications . Controlled, population-based studies are needed to assess the risk of BCG in HIV-infected children. Proc Natl Acad Sci U S A, 2003 Oct 14, 100(21), 12420 - 5 Epub 2003 Oct 13. The primary mechanism of attenuation of bacillus Calmette-Guerin is a loss of secreted lytic function required for invasion of lung interstitial tissue; Hsu T et al.; Tuberculosis remains a leading cause of death worldwide, despite the availability of effective chemotherapy and a vaccine . Bacillus Calmette-Guerin (BCG), the tuberculosis vaccine, is an attenuated mutant of Mycobacterium bovis that was isolated after serial subcultures, yet the functional basis for this attenuation has never been elucidated . A single region (RD1), which is abse