Korean J Intern Med, 2002 Mar, 17(1), 45 - 50 Electron microscopic evaluation of adhesion of Helicobacter pylori to the gastric epithelial cells in chronic gastritis; Chun HJ et al.; BACKGROUND: The adhesion of H . pylori to the gastric epithelial cells may be an essential step for the pathophysiology of various H . pylori-induced gastrointestinal diseases . The purpose of this study was to investigate the ultrastructural relation of H . pylori and gastric epithelial cells in their adhesion . METHODS: Endoscopic biopsy of gastric antrum and body was performed from 15 patients (9 men, 6 women) with chronic gastritis and H . pylori infection . The specimens were processed for electron microscopy and observed with a transmission electron microscope (Hitachi H-600) . RESULTS: On the basis of morphological appearances, the adhesions of H . pylori to the gastric epithelial cells were categorized into three types; filamentous connection, adhesion pedestals and membrane fusion . Coccoid and undetermined forms adhered mainly by the filamentous connection, whereas the bacillary forms adhered primarily by the adhesion pedestals and membrane fusion . CONCLUSION: Various types of adhesion were associated with H . pylori and gastric epithelium . Further studies are needed to evaluate the influence of different types of adhesion to the pathophysiology of H . pylori. Bull Soc Pathol Exot, 2002 Mar, 95(1), 34 - 6 {Bacillary angiomatosis in an adult infected with HIV-1 at an early stage of immunodepression in Abidjan, Côte d'Ivoire}; Minga KA et al.; Human immunodeficiency virus (HIV)-associated bacillary angiomatosis has rarely been described in Africa . We report here the first case in Cote d'Ivoire . Although in industrialised countries bacillary angiomatosis has been described in patients with low CD4 count, this episode occurred in the first year following HIV-seroconversion in an adult patient with more than 500 CD4 cells per cubic millimetre . Symptoms rapidly and totally disappeared under erythromycin treatment, although with a relapse two years after the end of the first episode . In Africa where people living with HIV often present chronic cutaneous lesions, bacillary angiomatosis may be under-diagnosed . Bacillary angiomatosis must be systematically considered in face of lesions similar to Kaposi's sarcoma . Improving knowledge on symptoms of bacillary angiomatosis in Africa should lead to better treatment and a better estimation of its true frequency which may be underestimated. Cancer Immunol Immunother, 2002 Jun, 51(4), 200 - 8 Epub 2002 Apr 09. Anti-metastatic activity of hapten-modified autologous tumor cell vaccine in an animal tumor model; Sojka DK et al.; We used mice from which the primary 410.4 mammary carcinoma had been surgically excised to assess the anti-metastatic activity of low-dose cyclophosphamide (CY) followed by vaccination with dinitrophenyl (DNP)-modified, irradiated, autologous tumor cells (ATC) admixed with bacille Calmette-Guerin (BCG) . Our studies revealed that CY treatment of mice followed by vaccination with DNP-modified . irradiated, ATC admixed with BCG improved the relapse-free survival compared to the survival of mice receiving either CY followed by vaccination with unmodified, irradiated, ATC admixed with BCG, or saline (control group) . In addition, our studies demonstrated the importance of CY administration in eliciting the therapeutic effect of DNP-modified ATC vaccine against metastatic disease . The therapeutic effect of CY followed by DNP-modified ATC vaccine was abrogated by depletion of CD4(+) or CD8(+) T-cells, illustrating the importance of both T-cell subsets for the anti-metastatic effect of this therapeutic protocol . In addition, neutralizing anti-IFN-gamma monoclonal antibody (mAb), or neutralizing anti-tumor necrosis factor (TNF) mAb reduced the relapse-free survival of mice treated with CY followed by DNP-modified ATC vaccine, indicating the importance of both cytokines for the realization of the anti-metastatic effect of this therapeutic protocol . Since the therapeutic protocol used in our studies was similar to that employed by Berd et al . as postsurgical adjuvant therapy in cancer patients and yielded a comparable anti-metastatic effect, the information obtained from the current studies with our clinically relevant experimental tumor model is expected to shed light on the mechanism(s) by which the anti-metastatic effect of this post-surgical adjuvant therapy is realized in cancer patients. Med Sci Monit, 2002 May, 8(5), BR168 - 71 Proteinases from Bacillus intermedius secreted in the late stages of sporulation; Balaban NP et al.; BACKGROUND: Proteinases are widely used in various fields of medicine, such as the treatment of burns, purulent wounds, or decubitus ulcers . On the basis of new microbial proteinases produced by nonpathogenic organisms, a new generation of medical preparations can be developed . Representatives of the Bacillus genera are nonpathogenic and are suitable for producing various proteases in large quantities . B . intermedius is shown to produce a set of alkaline proteases at the early and late stationary phase of growth . MATERIAL/METHODS: The activity of alkaline proteinases was determined using synthetic chromogenous substrates Z-Glu-pNA and Z-Ala-Ala-Leu-pNA . To determine beta-galactosidase activity, 2-nitro-beta-D-galactopyranosid was used . Spores were calculated by phase-contrast microscopy . RESULTS: During the late stages of sporulation B . intermedius 3-19 cells were shown to secrete two proteinases into the medium: glutamyl endopeptidase, with maximum activity at 40 hours of growth, and subtilisin, with maximum activity at 44 hours of growth . Evidence for the secretion of these enzymes into the medium was provided by measuring beta-galactosidase activity . CONCLUSIONS: Our results show that proteinases from B . intermedius (glutamyl endopeptidase 2 and subtilisin 2) in the late stationary phase of growth are secreted enzymes . This suggests that these enzymes play a role in sporulation. Infect Immun, 2002 Jun, 70(6), 3026 - 32 Correlation of ESAT-6-specific gamma interferon production with pathology in cattle following Mycobacterium bovis BCG vaccination against experimental bovine tuberculosis; Vordermeier HM et al.; Vaccine development and the understanding of the pathology of bovine tuberculosis in cattle would be greatly facilitated by the definition of immunological correlates of protection and/or pathology . To address these questions, cattle were vaccinated with Mycobacterium bovis bacillus Calmette-Guerin (BCG) and were then challenged with virulent M . bovis . Applying a semiquantitative pathology-scoring system, we were able to demonstrate that BCG vaccination imparted significant protection by reducing the disease severity on average by 75% . Analysis of cellular immune responses following M . bovis challenge demonstrated that proliferative T-cell and gamma interferon (IFN-gamma) responses towards the M . bovis-specific antigen ESAT-6, whose gene is absent from BCG, were generally low in vaccinated animals but were high in all nonvaccinated calves . Importantly, the amount of ESAT-6-specific IFN-gamma measured by enzyme-linked immunosorbent assay after M . bovis challenge, but not the frequency of responding cells, correlated positively with the degree of pathology found 18 weeks after infection . Diagnostic reagents based on antigens not present in BCG, like ESAT-6 and CFP-10, were still able to distinguish BCG-vaccinated, diseased animals from BCG-vaccinated animals without signs of disease . In summary, our results suggest that the determination of ESAT-6-specific IFN-gamma, while not a direct correlate of protection, constitutes nevertheless a useful prognostic immunological marker predicting both vaccine efficacy and disease severity. Int J Urol, 2002 Apr, 9(4), 219 - 24 Immunological protection induced by bacillus Calmette-Guérin treatment in a murine bladder tumor model; Iwasaki A et al.; BACKGROUND: It has been previously reported that MBT-2 tumor growth is completely inhibited when mice are inoculated with bacillus Calmette-Guerin (BCG) . In this study it was examined whether or not vaccination with a mixture of BCG and MBT-2 cells also induces immunological protection against murine bladder tumors . METHODS: Seven hundred thousand MBT-2 cells and 1 mg of BCG (Tokyo 172 strain) per mouse were injected subcutaneously into female C3H/HeN mice . Four and eight weeks after vaccination with this mixture, animals were reinoculated with MBT-2 cells alone or MBT-2 cells cocultured with BCG . RESULTS: Animals vaccinated with a mixture of BCG and MBT-2 cells showed MBT-2 tumor growth but completely rejected the MBT-2 cells cocultured with BCG . MBT-2 cells cocultured with BCG developed into tumors when they were inoculated into the control animals . Splenocytes prepared from vaccinated animals showed specific cytocidal activity against MBT-2 cells precultured with BCG . CONCLUSIONS: The results suggest that a mixture of BCG and MBT-2 cells induces antitumor immunological protection against BCG- or MBT-2-associated antigens presented on MBT-2 cells precultured with BCG. Biochem J, 2002 Aug 15, 366(Pt 1), 281 - 8 Cysteine-less glycosylphosphatidylinositol-specific phospholipase C is inhibited competitively by a thiol reagent: evidence for glyco-mimicry by p-chloromercuriphenylsulphonate; Stanton JD et al.; Glycosylphosphatidylinositol (GPI)-specific phospholipases are highly valuable for studying the structure and function of GPIs . GPI-specific phospholipase C (GPI-PLC) from Trypanosoma brucei and phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus are the most widely studied of this class of phospholipases C . Inhibition of protein activity by thiol reagents is indicative of the participation of cysteine residues in biochemical events . The thiol reagent p-chloromercuriphenylsulphonate (pCMPS) inhibits T . brucei GPI-PLC, which has eight cysteine residues . Surprisingly, we found that the activity of B . cereus PI-PLC is also blocked by pCMPS, although the protein does not contain cysteine residues . Inhibition of B . cereus PI-PLC was reversed when pCMPS was size-separated from a preformed pCMPS.PI-PLC complex . In contrast, no activity was recovered when T . brucei GPI-PLC was subjected to a similar protocol . Equimolar beta-mercaptoethanol (beta-ME) reversed the inhibition of PI-PLC activity in a pCMPS.PI-PLC complex . For T . brucei GPI-PLC, however, ultrafiltration of the pCMPS.GI-PLC complex and addition of a large excess of beta-ME was necessary for partial recovery of enzyme activity . Thus T . brucei GPI-PLC is susceptible to inactivation by covalent modification with pCMPS, whereas PI-PLC is not . Kinetic analysis indicated that pCMPS was a competitive inhibitor of PI-PLC when a GPI was a substrate . Curiously, with phosphatidylinositol as substrate, inhibition was no longer competitive . These data suggest that pCMPS is a glyco-mimetic that occupies the glycan binding site of PI-PLC, from where, depending on the substrate, it inhibits catalysis allosterically or competitively. J Agric Food Chem, 2002 May 22, 50(11), 3309 - 16 Modification of ascorbic acid using transglycosylation activity of Bacillus stearothermophilus maltogenic amylase to enhance its oxidative stability; Bae HK et al.; Ascorbic acid (1), a natural antioxidant, was modified by employing transglycosylation activity of Bacillus stearothermophilus maltogenic amylase with maltotriose and acarbose as donor molecules to enhance its oxidative stability . The transglycosylation reaction with maltotriose as donor created mono- and di-glycosyl transfer products with an alpha-(1,6)-glycosidic linkage . In addition, two acarviosine-glucosyl transfer products were generated when transglycosylation was performed with acarbose as a donor . All transfer products were observed by TLC and HPLC, and purified by Q-sepharose anion exchange and Biogel P-2 gel permeation chromatographies . LC/MS and (13)C NMR analyses revealed that the structures of the transfer products were 6-O-alpha-D-glucosyl- (2) and 6-O-alpha-D-maltosyl-ascorbic acids (3) in the reaction of maltotriose, and 6-O-alpha-acarviosine-D-glucosyl- (4) and 2-O-alpha-acarviosine-D-glucosyl ascorbic acids (5) in the reaction of acarbose . The stability of the transglycosylated ascorbic acid derivatives was greatly enhanced against oxidation by Cu(2+) ion and ascorbate oxidase . Among them, compound 3 proved to be the most stable against in vitro oxidation . The antioxidant effects of glycosyl-derivatives of ascorbic acid on the lipid oxidation in cooked chicken breast meat patties indicated that they had antioxidant activities similar to that of ascorbic acid . It is suggested that the transglycosylated ascorbic acids can possibly be applied as effective antioxidants with improved stability in food, cosmetic, and other applications. Mol Immunol, 2002 May, 38(12-13), 903 - 9 Inherited disorders of IL-12- and IFNgamma-mediated immunity: a molecular genetics update; Doffinger R et al.; In the last 6 years, considerable advances have been made in the molecular analysis of a rare clinical syndrome: Mendelian susceptibility to mycobacterial disease (MSMD) . Infection with poorly virulent environmental non-tuberculous mycobacteria (NTM) or vaccination with bacillus Calmette-Guerin (BCG) may cause disseminating and even fatal disease in individuals suffering from this syndrome . Mutations in five genes (IFNGR1, IFNGR2, STAT1, IL12B and IL12RB1) have been shown to be responsible for MSMD and further allelic heterogeneity accounts for the existence of nine distinct inherited disorders . All of these disorders are caused by impaired IFNgamma-mediated immunity . These results have important medical and biological implications . In this report, we update the disease-causing mutations reported in the literature. Dis Colon Rectum, 2002 Apr, 45(4), 502 - 7 Activity of HspE7, a novel immunotherapy, in patients with anogenital warts; Goldstone SE et al.; PURPOSE: Human papillomavirus causes anogenital squamous intraepithelial lesions, warts, and cancer . Treatment of squamous intraepithelial lesions to prevent cancer often requires extensive surgery . We tested a human papillomavirus-specific immunotherapy, HspE7, as a potential alternative . METHODS: HspE7 was constructed by fusing heat shock protein Hsp65 from bacille Calmette-Guerin to E7 protein from human papillomavirus-16 . Improvement in pathologic diagnosis of patients with persistent high-grade squamous intraepithelial lesions was studied in an open-label trial (HspE7 500 microg monthly x3) . Anogenital warts were not a trial parameter, but a retrospective review of the medical records of the first 22 patients enrolled at one site was undertaken to estimate the quality and frequency of responses of anogenital warts . Patients with warts by physical examination at baseline were scored at 24 weeks as to the percent reduction in wart size . RESULTS: Fourteen of the 22 patients had warts at baseline . At Week 24, 3 of the 14 patients had complete resolution of their warts, and 10 had warts reduced in size an estimated 70 to 95 percent . The remaining patient's warts increased in size . The reduction in size in most patients greatly diminished the procedure necessary for complete ablation . No serious or severe adverse events were related to HspE7 . CONCLUSIONS: A retrospective review of patients' medical records suggests that HspE7 may be broadly active in anogenital warts . This activity crosses multiple human papillomavirus types . The warts improved substantially but usually did not totally disappear within six months . Patient follow-up continues . A new randomized, placebo-controlled trial is underway to evaluate these findings. Mol Cell Probes, 2002 Feb, 16(1), 41 - 8 In situ identification of mycobacteria in Crohn's disease patient tissue using confocal scanning laser microscopy; Naser SA et al.; The diversity in the methodology employed to investigate Crohn's disease (CD) etiology has added significantly to the controversy of the mycobacterial role in this chronic inflammatory bowel disease . Mycobacterium avium subsp paratuberculosis (MAP), a proposed and suspected agent in many CD patients, is a fastidious and very slow grower bacillus, which causes Johne's disease (JD) in cattle . The methodology that has been widely and successfully used for isolation and identification of MAP from and in JD animals is not reliable and has proven to be unsuccessful in achieving the same objectives for CD diagnosis . In this study, a Confocal Scanning Laser Microscopy (CSLM) system has been employed in an attempt to detect MAP in CD patient . In situ hybridization was performed on full thickness tissue using rabbit anti-MAP polyclonal antibody that was adsorbed with E . coli protein extracts . Consequently, MAP was detected in the microvilli region in tissue specimens from CD patient and not in the controls . In the same CD tissue specimen, MAP was not detected when isotype normal rabbit sera was employed . The polyclonal antibody marker may be replaced with monoclonal antibodies, if available, or with MAP-specific-DNA or RNA probes . This technique adds an additional approach to investigate MAP role in CD etiology especially when the culture approach is long and inconsistent. Structure (Camb), 2002 Mar, 10(3), 311 - 7 Structure and dynamics of the anticodon arm binding domain of Bacillus stearothermophilus Tyrosyl-tRNA synthetase; Guijarro JI et al.; The structure of a recombinant protein, TyrRS(delta4), corresponding to the anticodon arm binding domain of Bacillus stearothermophilus tyrosyl-tRNA synthetase, has been solved, and its dynamics have been studied by nuclear magnetic resonance (NMR) . It is the first structure described for such a domain of a tyrosyl-tRNA synthetase . It consists of a five-stranded beta sheet, packed against two alpha helices on one side and one alpha helix on the other side . A large part of the domain is structurally similar to other functionally unrelated RNA binding proteins . The basic residues known to be essential for tRNA binding and charging are exposed to the solvent on the same face of the molecule . The structure of TyrRS(delta4), together with previous mutagenesis data, allows one to delineate the region of interaction with tRNATyr. Biosci Biotechnol Biochem, 2002 Mar, 66(3), 682 - 4 Requirement for C-terminal extension to the RNA binding domain for efficient RNA binding by ribosomal protein L2; Hayashi T et al.; Ribosomal protein L2 is a primary 23S rRNA binding protein in the large ribosomal subunit . We examined the contribution of the N- and C-terminal regions of Bacillus stearothermophilus L2 (BstL2) to the 23S rRNA binding activity . The mutant desN, in which the N-terminal 59 residues of BstL2 were deleted, bound to the 23S rRNA fragment to the same extent as wild type BstL2, but the mutation desC, in which the C-terminal 74 amino acid residues were deleted, abolished the binding activity . These observations indicated that the C-terminal region is involved in 23S rRNA binding . Subsequent deletion analysis of the C-terminal region found that the C-terminal 70 amino acids are required for efficient 23S rRNA binding by BstL2 . Furthermore, the surface plasmon resonance analysis indicated that successive truncations of the C-terminal residues increased the dissociation rate constants, while they had little influence on association rate constants . The result indicated that reduced affinities of the C-terminal deletion mutants were due only to higher dissociation rate constants, suggesting that the C-terminal region primarily functions by stabilizing the protein L2-23S rRNA complex. Biosci Biotechnol Biochem, 2002 Mar, 66(3), 516 - 22 Active form of dipteran-specific insecticidal protein cryllA produced by Bacillus thuringiensis subsp . israelensis; Yamagiwa M et al.; The nucleotide sequence of the cry11A gene from Bacillus thuringiensis subsp . israelensis strain HD522 was analyzed and the molecular characterization of CryllA toxin was done . The 70-kDa CryllA protoxin was processed in vitro into 36- and 32-kDa fragments by trypsin and into 34- and 32-kDa fragments by gut proteases from C . pipiens . These two processed fragments are associated together to form the heterodimer . The results of the binding assay with BBMV and the bioassay toward C . pipiens larvae suggested that the heterodimer was biologically as active as the non-digested CryllA toxin and the intramolecular cleavage did not promote the insecticidal activity . These results suggested that a probable complex of the 36- or 34-kDa and 32-kDa fragments was also one of the possible active forms of Cry11A, and that the biological functions of CryllA was not essentially affected by the intramolecular cleavage of the 70-kDa protein. Br Poult Sci, 2002 Mar, 43(1), 70 - 7 The potential for the improvement of the nutritive value of soya-bean meal by different proteases in broiler chicks and broiler cockerels; Ghazi S et al.; 1 . The potential for improving the nutritive value of commercial solvent-extracted, heat-treated soya-bean meal (SBM) by protease treatment was measured using growing broiler chicks and tube-fed broiler cockerels . 2 . SBM was pre-treated (50 degrees C for 2 h) with water alone; at alkaline pH (initial pH 8.25) with and without protease P1 (isolated from a Bacillus species) or at acid pH (initial pH 4.5) with and without protease P2 (isolated from an Aspergillus species) and incorporated into diets (290 g SBM/kg diet) for broiler chicks (20 chicks/treatment) . Only protease P2 treatment improved chick performance; from 7 to 28 d of age, chicks fed on treated SBM had greater feed intakes and gained more weight than chicks fed on untreated SBM . Both proteases P1 and P2 significantly reduced chick serum anti-soya antibodies while protease P2 treatment increased apparent ileal nitrogen (N) digestibility and apparent N retention across the whole digestive tract . 3 . Two tube-feeding experiments established that, of the treated SBMs used in experiment 1, only protease P2 treatment improved apparent N digestibility and true metabolisable energy . Also it was shown that increasing the temperature at which treated SBM was dried to 60 degrees C, compared with freeze-drying or drying at 50 degrees C reduced apparent N digestibility and true metabolisable energy of SBM with no significant interactions between enzyme treatment and drying temperature for both apparent N digestibility and TME . 4 . It is concluded that, overall, the nutritional value of SBM assayed in a growth trial and by tube feeding was improved by treatment with protease P2 and not by treatment with protease P1. Res Microbiol, 2002 Apr, 153(3), 165 - 71 Inhibition of 4-nitroquinoline-1-oxide genotoxicity by Bacillus strains; Caldini G et al.; The effect of 16 Bacillus strains from pharmaceutical probiotic preparations (Bacillus spp.) and collection (B . subtilis, B . firmus, B . megaterium, B . pumilus) on genotoxicity induced by the known mutagen 4-nitroquinoline-1-oxide (4-NQO) was studied using the short-term bacterial assay SOS-chromotest . with Escherichia coli PQ37 as the tester organism . It was found that the activity of 0.1 mM 4-NQO was reduced (P < 0.01) after coincubation with Bacillus suspensions (10(8) CFU/ml for 150 min at 37 degrees C) . All isolates showed potential for deactivating 4-NQO, and genotoxicity inhibition ranged from 92.9 to 100% . There were no appreciable differences in behaviour observed among probiotic and collection strains or in relation to species . The observed antigenotoxicity was associated with a clear-cut modification of 4-NQO molecular characteristics. J Vet Med B Infect Dis Vet Public Health, 2002 Mar, 49(2), 89 - 96 Diagnosis of Mycobacterium bovis infection in calves sensitized by mycobacteria of the avium/intracellulare group; Amadori M et al.; Because of the frequent exposure of cattle to mycobacteria of the avium/intracellulare group, an investigation was carried out into the possible repercussions thereof on the diagnosis of bovine tuberculosis . Three calves from a bovine tuberculosis-free herd, scored avian reactors in the gamma-interferon assay for bovine tuberculosis, were sedated and inoculated endotracheally with a virulent Mycobacterium bovis strain . Then, three other avian reactors were housed with the above donor calves . Mycobacterium bovis was isolated from the nasal swabs of the three endotracheally infected, donor calves . On these samples, TB complex-specific polymerase chain reaction (PCR) tests for IS6110 were also positive, albeit with a different time kinetics . The three contact-infected calves showed clear immunological signs of infection; however, their nasal swabs were always PCR-negative and only Mycobacterium avium was isolated . In the endotracheally infected donor calves there was a rise of the gamma-interferon responses to avian and bovine purified protein derivative (PPD) tuberculins, which reached the same stable plateau levels over the whole experiment . The above effect was also observed in the contact-infected calves, even though the response to avian PPD tuberculin always remained at a higher level . By using conventional bovine and avian PPD tuberculins, the comparative intradermal test was generally positive in endotracheally infected, as opposed to contact-infected calves; a positive intradermal test for M . bovis was obtained in two contact-infected calves by different bovine PPD tuberculins based on M . bovis bacillus Calmette-Guerin (BCG) secreted or somatic antigens . It was concluded that M . bovis infection may be concealed for some time in cattle sensitized by mycobacteria of the avium/intracellulare group and that different diagnostic procedures should be adopted for such animals. Environ Monit Assess, 2002 Apr, 75(2), 107 - 19 Effect of an industrial discharge on water quality and periphyton structure in a pampeam stream; Giorgi A et al.; Seasonal sampling was carried out at four sites on a pampean stream that receives industrial effluent from two textile factories . To evaluate water quality, several physical and chemical parameters were examined and the periphyton growing on cattail (Typha latifolia L.) were analyzed . Water quality and periphyton structure differed significantly between sites upstream and downstream of the discharge . Differences in temperature and also in concentrations of phosphate, dissolved oxygen, and phaeopigment were detected . At the same time, changes in the dominant algae groups were observed . Downstream of the industrial discharge, the number of Bacillariophyta decreased, while species of Cianophyta and Euglenophyta were more abundant . This abundance correlated with increased phosphate and organic matter content and decreased oxygen concentration . Although this study did not detect a reduction in the number of species, similarity between stands decreased downstream of the industrial discharge . Changes in community structure were readily detected in this situation because the communities of the polluted and unpolluted zones were qualitatively different . Periphyton growing naturally on Typha latifolia is a useful indicator of the impact of waste waters on the biota and can also be used to evaluate water body recovery. Eur Respir J, 2002 Apr, 19(4), 765 - 75 European framework for tuberculosis control and elimination in countries with a low incidence . Recommendations of the World Health Organization (WHO), International Union Against Tuberculosis and Lung Disease (IUATLD) and Royal Netherlands Tuberculosis Association (KNCV) Working Group; Broekmans JF et al.; As countries approach the elimination phase of tuberculosis, specific problems and challenges emerge, due to the steadily declining incidence in the native population, the gradually increasing importance of the importation of latent tuberculosis infection and tuberculosis from other countries and the emergence of groups at particularly high risk of tuberculosis . Therefore, a Working Group of the World Health Organization (WHO), the International Union Against Tuberculosis and Lung Disease (IUATLD) and the Royal Netherlands Tuberculosis Association (KNCV) have developed a new framework for low incidence countries based on concepts and definitions consistent with those of previous recommendations from WHO/IUATLD Working Groups . In low-incidence countries, a broader spectrum of interventions is available and feasible, including: 1) a general approach to tuberculosis which ensures rapid detection and treatment of all the cases and prevention of unnecessary deaths; 2) an overall control strategy aimed at reducing the incidence of tuberculosis infection (risk-group management and prevention of transmission of infection in institutional settings) and 3) a tuberculosis elimination strategy aimed at reducing the prevalence of tuberculosis infection (outbreak management and provision of preventive therapy for specified groups and individuals) . Government and private sector commitment towards elimination, effective case detection among symptomatic individuals together with active case finding in special groups, standard treatment of disease and infection, access to tuberculosis diagnostic and treatment services, prevention (e.g . through screening and bacille Calmette-Gueria immunization in specified groups), surveillance and treatment outcome monitoring are prerequisites to implementing the policy package recommended in this new framework document. J Am Mosq Control Assoc, 2002 Mar, 18(1), 65 - 7 An isolate of Bacillus circulans toxic to mosquito larvae; Darriet F et al.; A new strain of Bacillus circulans isolated from a larva of Culex quinquefasciatus showed larvicidal activity on 3 mosquitoes of medical importance . Compared to Bacillus sphaericus strain 2362, this B . circulans isolate proved less toxic to Cx . quinquefasciatus and Anopheles gambiae but was 107 times more toxic to Aedes aegypti . Moreover, in comparison to other studies, B . circulans was at least as pathogenic as B . thuringiensis var . israelensis in Ae . aegypti . The tests have showed that the toxicity of the bacterial culture of B . circulans resulted from its spores and not from the insecticidal effect of chitinases or exotoxins. J Am Mosq Control Assoc, 2002 Mar, 18(1), 57 - 62 Sterilization of Bacillus thuringiensis israelensis products by gamma radiation; Becker N; This study examines the effect of routine gamma radiation based on cobalt 60 on the viability, mosquito larvicidal activity, and density of bacillus spores in the soil . Although 1 g of unirradiated powder of Bacillus thuringiensis israelensis (Bti) contains on average 6.2 x 10(9) spores, no spores survived radiation doses of 20.6 kGy and higher . Radiation at a dose of 20-25 kGy caused a 20-30% reduction in the effectiveness of Bti powder against mosquito larvae . In areas treated with unirradiated Bti material on average twice a year, soil contained 700,000 to 44 million spores per gram . In areas treated with irradiated Bti products, either no Bti spores or fewer than 100,000 were found per gram of soil . A radiation dose of 25 kGy fulfills the requirements of killing all spores in a Bti product and maintaining the effectiveness of the product in routine treatments . No viable spores remain in water used for household purposes or irrigation of garden areas when irradiated Bti fizzy tablets are used in water containers . Irradiation of Bti products fulfills the requirements of drinking water regulations and thus allows these products to be used widely. J Am Mosq Control Assoc, 2002 Mar, 18(1), 52 - 6 Indoor thermal fogging against vector mosquitoes with two Bacillus thuringiensis israelensis formulations, Vectobac ABG 6511 water-dispersible granules and Vectobac 12AS liquid; Yap HH et al.; Bioefficacy of thermal fogging application of 2 Bacillus thuringiensis israelensis formulations, Vectobac ABG 6511 water-dispersible granules (3,000 international toxic unit {ITU}/mg) and Vectobac 12AS liquid (1,200 ITU/mg), was assessed for larvicidal activities against Aedes aegypti, Aedes albopictus, Anopheles dirus, and Culex quinquefasciatus . Portable Agrofog AF35 sprayers were used to apply the 2 formulations indoors in residential premises on Penang Island, Malaysia . Vectobac ABG 6511 showed good larvicidal effect against all 4 mosquito species at 3 of the higher doses tested (2.91 x 10(9), 1.45 x 10(9), and 0.71 x 10(9) ITU/ha), with more than 96% mortality at 48 h after spraying . As a comparative formulation, Vectobac 12AS also showed good larvicidal activity against all 4 mosquito species at 2 of the higher doses tested (2.87 x 10(9) and 1.46 x 10(9) ITU/ha), with more than 92.5% mortality at 48 h after spraying . Larvae of An . dirus were significantly more susceptible to both water-based Vectobac formulations when compared to the other 3 mosquito species . Both microbial formulations showed better efficacy at higher doses . However, even at the lowest dose tested, Vectobac ABG 6511 and Vectobac 12AS (both at 0.36 x 10(9) ITU/ha) showed larvicidal properties, with more than 66% mortality at 48 h after spraying . Overall, for this bacterial agent, the water-dispersible granule formulation has better prospects than the liquid formulation for the control of larvae of vector mosquitoes. Am Fam Physician, 2002 Apr 15, 65(8), 1599 - 604 Recognition and management of acute pesticide poisoning; Simpson WM Jr et al.; Most poisonings from pesticides do not have a specific antidote, making decontamination the most important intervention . For maximal benefit to the patient, skin, eye, and gastric decontamination should be undertaken while specifics of the poisoning are being determined . As in most illnesses and injuries, the history of the poisoning is of great importance and will determine specific needs for decontamination and therapy, if any exist . Protection of health care workers during the decontamination process is important and frequently overlooked . Skin decontamination is primarily accomplished with large volumes of water, soap, and shampoo . Gastric decontamination by lavage is indicated if ingestion of the poisoning has occurred within 60 minutes of patient presentation . Activated charcoal, combined with a cathartic, is also indicated in most poisonings presenting within 60 minutes of ingestion . With large volume ingestion poisonings, activated charcoal may be used after 60 minutes, but little data exist to support this practice . Syrup of ipecac is no longer recommended for routine use . The cholinergic syndrome "all faucets on" characterizes poisoning by organophosphates and carbamates . Organochlorine insecticides (lindane and other treatments for scabies and lice) can produce seizures with excessive use or use on large areas of nonintact skin . Non-dipyridyl herbicides, biocides (including pyrethrins, pyrethroids, and Bacillus thuringiensis) rarely produce anything other than mild skin, eye, and/or gastrointestinal irritation on topical exposure or ingestion. Tidsskr Nor Laegeforen, 2002 Mar 10, 122(7), 708 - 9 {The discovery of the leprosy bacillus}; Irgens LM; Gerhard Henrik Armauer Hansen (1841-1912) worked on leprosy throughout his career . Following his discovery of the leprosy bacillus in 1873, he proposed legislation that, when enacted in 1877 and 1885, established preventive measures aimed at isolating infectious patients . Around 1920, leprosy was more or less eradicated in Norway after a period of decline starting in 1850 . Over this period, more than 8,000 cases were registered . Armauer Hansen's unique research achievement was based on a scientific and medical infrastructure in place long before he started his work . This context had several implications, though the discovery of the leprosy bacillus holds a particular fascination, with bearings on the interaction between medicine and the community even today. Rev Panam Salud Publica, 2002 Mar, 11(3), 166 - 71 {A mathematical model of the annual risk of tuberculosis infection in Cali, Colombia}; de la Pava E et al.; OBJECTIVE: The annual risk of infection (ARI) for tuberculosis is the probability that an individual without previous contact with the tubercle bacillus has of being infected during the course of a year . The ARI is the most appropriate indicator for estimating the degree of tuberculosis infection in a population . The objective of this paper was to estimate the ARI and its trends in the city of Cali, Colombia, using data provided by the Municipal Secretariat of Health . METHODS: We used a deterministic model of the dynamics of pulmonary tuberculosis . The flows among the population subgroups were based on the natural history of the disease, taking vaccination into account . Using the data from the Municipal Secretariat of Health, we estimated the initial conditions and the values of the parameters . RESULTS: The mean ARI values were 1.24% in the 1970s, 0.93% in the 1980s, and 0.85% in the 1990s . In order to assess trends, we attempted to predict the annual risk, utilizing a nonlinear least-squares adjustment of the data on the overall percentage for each year . With that approach, we projected that the ARI in 2003 would be 1.3%, indicating a return to the patterns found in the 1970s . CONCLUSIONS: The estimated risk of tuberculosis infection in Cali during the decades of the 1970s, the 1980s, and the 1990s was very high in comparison with the risk in countries such as the Netherlands, which in 1985 had an ARI of 0.012% . However, the ARI in Cali is not so high in comparison to indices for other countries of South America, which range from 0.5% to 1.5% . This model and the simulation it produced showed a rising trend in the ARI for Cali, as well as demonstrated that the ARI will tend to continue to rise if control measures are not improved. Biochim Biophys Acta, 2002 Apr 12, 1561(2), 171 - 9 Amino acid and divalent ion permeability of the pores formed by the Bacillus thuringiensis toxins Cry1Aa and Cry1Ac in insect midgut brush border membrane vesicles; Kirouac M et al.; The pores formed by Bacillus thuringiensis insecticidal toxins have been shown to allow the diffusion of a variety of monovalent cations and anions and neutral solutes . To further characterize their ion selectivity, membrane permeability induced by Cry1Aa and Cry1Ac to amino acids (Asp, Glu, Ser, Leu, His, Lys and Arg) and to divalent cations (Mg(2+), Ca(2+) and Ba(2+)) and anions (SO(4)(2-) and phosphate) was analyzed at pH 7.5 and 10.5 with midgut brush border membrane vesicles isolated from Manduca sexta and an osmotic swelling assay . Shifting pH from 7.5 to 10.5 increases the proportion of the more negatively charged species of amino acids and phosphate ions . All amino acids diffused well across the toxin-induced pores, but, except for aspartate and glutamate, amino acid permeability was lower at the higher pH . In the presence of either toxin, membrane permeability was higher for the chloride salts of divalent cations than for the potassium salts of divalent anions . These results clearly indicate that the pores are cation-selective. FEBS Lett, 2002 May 8, 518(1-3), 79 - 82 Action pattern and subsite mapping of Bacillus licheniformis alpha-amylase (BLA) with modified maltooligosaccharide substrates; Kandra L et al.; This study represents the first characterisation of the substrate-binding site of Bacillus licheniformis alpha-amylase (BLA) . It describes the first subsite map, namely, number of subsites, apparent subsite energies and the dual product specificity of BLA . The product pattern and cleavage frequencies were determined by high-performance liquid chromatography, utilising a homologous series of chromophore-substituted maltooligosaccharides of degree of polymerisation 4-10 as model substrates . The binding region of BLA is composed of five glycone, three aglycone-binding sites and a 'barrier' subsite . Comparison of the binding energies of subsites, which were calculated with a computer program, shows that BLA has similarity to the closely related Bacillus amyloliquefaciens alpha-amylase. Appl Biochem Biotechnol, 2002 Feb, 97(2), 123 - 33 Characterization of a keratinolytic metalloprotease from Bacillus sp . SCB-3; Lee H et al.; A keratinolytic protease-producing microorganism was isolated from soybean paste waste and was identified as a strain of Bacillus sp . The keratinase was purified by polyethylene glycol precipitation and two successive column chromatographies with DEAE-Toyopearl 650C and Sephacryl S-200 HR . The purified enzyme had overall 11 purification folds with an 18% yield . The results of sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration on Sephacryl G-200 indicated that the purified enzyme was monomeric and had a molecular weight of 134 kDa . The optimum temperature and pH were 40 degrees C and 7.0, respectively . This enzyme was completely inhibited by EDTA and EGTA, and it was restored by the addition of Ca2+ and Mg2+ . These results suggested that it is a metalloprotease . The stimulated enzyme activity by reducing agents indicated that the reducing condition was important in the expression of the activity. Sao Paulo Med J, 2002 Mar 7, 120(2), 55 - 8 ABO, Lewis, secretor and non-secretor phenotypes in patients infected or uninfected by the Helicobacter pylori bacillus; de Mattos LC et al.; CONTEXT: Epidemiological studies have demonstrated higher frequencies of the O blood group and the non-secretor phenotype of ABH antigens among patients suffering from peptic ulcers . Since Helicobacter pylori has been established as the main etiological factor in this disease, controversies about the associations of the ABO and Lewis blood group phenotypes and secretor and non-secretor phenotypes in relation to susceptibility towards infection by this bacillus have been presented . OBJECTIVE: To verify the frequencies of ABO, Lewis blood group phenotypes, secretor and non-secretor phenotypes in patients infected or uninfected by H . pylori . DESIGN: Cross-sectional study . SETTING: Outpatient clinic . PARTICIPANTS: One hundred and twenty patients with dyspeptic symptoms who underwent endoscopy . MAIN MEASUREMENTS: ABO and Lewis blood group phenotypes were determined by a standard hemagglutination test and the secretor and non-secretor phenotypes were evaluated by saliva samples using the inhibitor hemagglutination test . RESULTS: The diagnosis of infection, made via breath and urea tests and confirmed using polymerase chain reaction (PCR) in gastric biopsy fragments, showed the presence of H . pylori in 61.7% of the patients and absence in 38.3% . The differences between the frequencies of the ABO blood group phenotypes among infected (A 27.0%; B 12.2%; AB 4.0% and O 56.8%) and uninfected patients (A 58.7%; B 13.0%; AB 4.3% and O 24.0%) were significant . The Lewis blood type, secretor and non-secretor phenotypes showed homogeneous distribution between the groups of patients analyzed . CONCLUSIONS: Our results suggest that the infection of H . pylori can be related to ABO blood groups but not to the Lewis blood group nor to secretor and non-secretor phenotypes. Biochemistry, 2002 May 14, 41(19), 6193 - 201 Pyrococcus furiosus alpha-amylase is stabilized by calcium and zinc; Savchenko A et al.; The hyperthermophilic archeon Pyrococcus furiosus produces an extracellular alpha-amylase that belongs to glycosyl hydrolases' family 13 . This enzyme is more thermostable than its bacterial and archaeal homologues (e.g., Bacillus licheniformis TAKA-term and Pyrococcus kodakaraensis KOD1 alpha-amylases, respectively) even without adding Ca(2+) ions . Unlike the TAKA-therm amylase that contains no cysteine, the P . furiosus enzyme contains five cysteines (C152, C153, C165, C387, and C430), only four of which (C152, C153, C387, and C430) are conserved in the P . kodakaraensis alpha-amylase . To test the potential function of cysteines in P . furiosus alpha-amylase stability, these five residues were substituted with Ser or Ala-either one-by-one or in sequence-to produce eight mutant enzymes . Mutation C165S dramatically destabilized P . furiosus alpha-amylase . At the same time, the quadruple mutant enzyme C152S/C153S/C387S/C430A (mutant SSCSA) was as thermostable as the wild-type enzyme . Mutant SSCSA and wild-type alpha-amylases were strongly destabilized by dithiothreitol and ethylenediaminetetraacetic acid, suggesting that metal binding can be involved in this enzyme's thermostability . Inductively coupled plasma-atomic emission spectrometry showed the presence of Ca(2+) and Zn(2+) metal ions in P . furiosus alpha-amylase . Although Ca(2+) is known to contribute to alpha-amylase's stability, the absence of two out of the three conserved Ca(2+) ligands in the P . furiosus enzyme suggests that a different set of amino acids is involved in this enzyme's Ca(2+) binding . We also provide evidence suggesting that Cys165 is involved in Zn(2+) binding and that Cys165 is essential for the stability of P . furiosus alpha-amylase at very high temperatures. Biochemistry, 2002 May 14, 41(19), 6178 - 84 Role of helix 3 in pore formation by the Bacillus thuringiensis insecticidal toxin Cry1Aa; Vachon V et al.; Helix 3 of the Cry1Aa toxin from Bacillus thuringiensis possesses eight charged amino acids . These residues, with the exception of those involved in intramolecular salt bridges (E90, R93, E112, and R115), were mutated individually either to a neutral or to an oppositely charged amino acid . The mutated genes were expressed, and the resultant, trypsin-activated toxins were assessed for their toxicity to Manduca sexta larvae and their ability to permeabilize M . sexta larval midgut brush border membrane vesicles to KCl, sucrose, raffinose, potassium gluconate, and N-methyl-D-glucamine hydrochloride with a light-scattering assay based on osmotic swelling . Most mutants were considerably less toxic than Cry1Aa . Replacing either E101, E116, E118, or D120 by cysteine, glutamine, or lysine residues had only minor effects on the properties of the pores formed by the modified toxins . However, half of these mutants (E101C, E101Q, E101K, E116K, E118C, and D120K) had a significantly slower rate of pore formation than Cry1Aa . Mutations at R99 (R99C, R99E, and R99Y) resulted in an almost complete loss of pore-forming ability . These results are consistent with a model in which alpha-helix 3 plays an important role in the mechanism of pore formation without being directly involved in determining the properties of the pores. Eur J Immunol, 1975 Oct, 5(10), 729 - 31 A leukocyte migration inhibition assay technique using blood clot fragments in vitro; Sandru G; Concomitantly with the capillary tube method, an in vitro leukocyte migration inhibition assay technique using blood clot fragments was performed in chicken egg albumin, Bacillus Calmette Guerin and complete Freund's adjuvant-treated immune guinea pigs . Results of both methods were concordant. J Infect Dis, 2002 May 15, 185(10), 1468 - 75 Epub 2002 Apr 30. Clinical and genetic heterogeneity of inherited autosomal recessive susceptibility to disseminated Mycobacterium bovis bacille calmette-guérin infection; Elloumi-Zghal H et al.; Five patients from 4 unrelated Tunisian families who presented with disseminated neonatal infection by Mycobacterium bovis bacille Calmette-Guerin strain were investigated . Two unrelated patients had different homozygous interleukin-12 receptor beta1 subunit gene splice-site mutations (64+5G-->A and 550-2A-->G) . Two siblings and 1 unrelated patient, all of whom were from the same town, carried the same mutation (297del8) within the interleukin-12p40 gene . This is the first description of familial cytokine deficiency reported so far . All patients had complete lack of expression of the affected polypeptide and a profound deficiency of in vitro interferon-gamma production . The clinical severity of the mycobacterial infection was heterogeneous, even among affected members of the same family, which suggests the intervention of modifying genes. J Urol, 2002 Jun, 167(6), 2408 - 12 Treatment of carcinoma in situ with intravesical bacillus Calmette-Guerin without maintenance; Griffiths TR et al.; PURPOSE: Data concerning the relative efficacy of intravesical bacillus Calmette-Guerin (BCG) on subgroups of carcinoma in situ of the bladder are limited . We report the outcome of primary carcinoma in situ and carcinoma in situ associated with Ta or T1 transitional cell carcinoma of the bladder treated with BCG . MATERIALS AND METHODS: Between 1987 and 1997, 135 patients (median age 70 years) with biopsy proven bladder carcinoma in situ underwent a standard course of 6 BCG instillations . Patients were divided into group 1-23 patients with primary carcinoma in situ, group 2-37 with carcinoma in situ associated with Ta transitional cell carcinoma and group 3-75 with carcinoma in situ associated with T1 transitional cell carcinoma . RESULTS: Median followup was 41 months . For groups 1 to 3, complete response rates at 3 months were 74% (17 of 23 cases), 70% (26 of 37) and 75% (56 of 75), respectively . The overall progression rates at 5 years were 20% (3 of 15 cases), 18% (4 of 22) and 49% (25 of 51) . Cancer specific survival rates were 83% (10 of 12 patients), 86% (12 of 14) and 59% (17 of 29), and the numbers of patients alive with the bladder intact were 60% (9 of 15), 58% (11 of 19) and 30% (12 of 40) . Patients in group 3 treated with BCG had progression significantly earlier than those in groups 1 and 2 (log-rank test p = 0.013) . A complete response to BCG in group 3 patients significantly delayed time to progression (Cox regression p = 0.001) but did not reduce death from transitional cell carcinoma . Indeed, only 38% (8 of 21) of complete responders were alive with the bladder intact at 5 years . CONCLUSIONS: A single course of BCG is remarkably effective for primary carcinoma in situ and carcinoma in situ associated with Ta transitional cell carcinoma but is suboptimal in patients with carcinoma in situ associated with T1 transitional cell carcinoma . Better outcomes in each of the 3 groups may have occurred with maintenance BCG. J Chromatogr A, 2002 Mar 15, 950(1-2), 131 - 7 Purification of a bacterial pullulanase on a fluidized bed of calcium alginate beads; Roy I et al.; Pullulanase from Bacillus acidopullulyticus was purified on a packed bed and a fluidized bed of calcium alginate beads . The binding of enzyme activity to the medium was found to follow Langmuir isotherm pattern . The maximum binding capacity was 1476 U ml(-1) matrix and the dissociation constant was 142 U ml(-1) . The dynamic binding capacities at 5% breakthrough in the packed and fluidized beds were 472 U ml(-1) and 644 U ml(-1), respectively . In the packed bed as well as the fluidized bed, an activity recovery of more than 95% with fold purification in the range of 46-59 was observed . The elution with a competitive inhibitor, viz . maltose, and high-fold purification indicate an affinity-based process . The purification process worked equally well with columns of bed volumes of 3.8 and 10 ml. Nat Prod Lett, 2002 Apr, 16(2), 101 - 6 Microbial transformation of danazol; Choudhary MI et al.; Danazol (17beta-hydroxy-17alpha-pregna-2,4-dien-20-yno-{2,3-d} isoxazole) (1) on fermentation with Fusarium lini, Aspergillus niger, and Cephalosporium aphidicola yielded 17beta-hydroxy-2-(hydroxymethyl)-17-alpha-pregn-4-en-20-yn-3-one (2) and 17beta-hydroxy-2-(hydroxymethyl)-17 alpha-pregna-1,4-dien-20-yn-3-one (3), while the fermentation of 1 with Bacillus cerus yielded compound 2 only. J AOAC Int, 2002 Mar-Apr, 85(2), 501 - 4 Optimized, one-step, recovery-enrichment broth for enhanced detection of Listeria monocytogenes in pasteurized milk and hot dogs; Knabel SJ; A one-step, recovery-enrichment broth, optimized Penn State University (oPSU) broth, was developed to consistently detect low levels of injured and uninjured Listeria monocytogenes cells in ready-to-eat foods . The oPSU broth contains special selective agents that inhibit growth of background flora without inhibiting recovery of injured Listeria cells . After recovery in the anaerobic section of oPSU broth, Listeria cells migrated to the surface, forming a black zone . This migration separated viable from nonviable cells and the food matrix, thereby reducing inhibitors that prevent detection by molecular methods . The high Listeria-to-background ratio in the black zone resulted in consistent detection of low levels of L . monocytogenes in pasteurized foods by both cultural and molecular methods, and greatly reduced both false-negative and false-positive results . oPSU broth does not require transfer to a secondary enrichment broth, making it less laborious and less subject to external contamination than 2-step enrichment protocols . Addition of 150mM D-serine prevented germination of Bacillus spores, but not the growth of vegetative cells . Replacement of D-serine with 12 mg/L acriflavin inhibited growth of vegetative cells of Bacillus spp . without inhibiting recovery of injured Listeria cells . oPSU broth may allow consistent detection of low levels of injured and uninjured cells of L . monocytogenes in pasteurized foods containing various background microflora. Can J Microbiol, 2002 Mar, 48(3), 262 - 7 Larvicidal toxins from Bacillus thuringiensis subspp . kurstaki, morrisoni (strain tenebrionis), and israelensis have no microbicidal or microbiostatic activity against selected bacteria, fungi, and algae in vitro; Koskella J et al.; The insecticidal toxins from Bacillus thuringiensis subspp . kurstaki (antilepidopteran), morrisoni strain tenebrionis (anticoleopteran), and israelensis (antidipteran) did not affect the growth of a variety of bacteria (8 gram-negative, 5 gram-positive, and a cyanobacterium), fungi (2 Zygomycetes, 1 Ascomycete, 2 Deuteromycetes, and 2 yeasts), and algae (primarily green and diatoms) in pure and mixed culture, as determined by dilution, disk-diffusion, and sporulation assays with purified free and clay-bound toxins . The insecticidal crystal proteins from B . thuringiensis subspp . kurstaki and israelensis had no antibiotic effect on various gram-positive bacteria. Can J Microbiol, 2002 Mar, 48(3), 256 - 61 Long-term survival and germination of Bacillus thuringiensis var . kurstaki in a field trial; Hendriksen NB et al.; Long-term survival, dispersal, and germination of Bacillus thuringiensis var . kurstaki DMU67R has been investigated in a field trial . An experimental cabbage plot was sprayed with DMU67R in 1993 and allowed to lie fallow since . The investigations reported here were carried out from 1997 to 2000 in this plot . High persistence of DMU67R for 7 years in the bulk soil of the plot has been demonstrated . The numbers have not significantly reduced since 1994, stabilizing around 6.6 x 10(2) cfu/g from 1996 to 2000 . Horizontal dispersal of DMU67R in the 1994-1999 period was limited . Vertical dispersal occurred from 1994 to 1999, as 77% of the population of DMU67R occurred in the 0-2 cm layer in 1994, while only 22% of the population was found there in 1999 . Most of the population in 1999 was present homogeneously in the upper 6 cm of the soil profile . Germination, as evidenced by the ratio of DMU67R cfu before and after heat treatment, was not observed in the bulk soil . However, in the rhizospheres of dandelion (Taraxacum officinalis) and quackgrass (Agropyron repens), 40 and 50% of DMU67R was present as vegetative germinated cells, respectively . No germination occurred in the rhizosphere of red fescue (Festuca rubra) . The material from the gut of the earthworm species Lumbricus rubellus, Lumbricus terrestris, and Apporrectodea caliginosa and from a tipulid larvae from the plot also contained vegetative cells of DMU67R . Further investigations of A . caliginosa showed that germination seemed to be restricted to the gut and that sporulation occurred after defecation . The germination of DMU67R in rhizospheres and in the gut of nontarget invertebrates suggests that survival in the soil of B . thuringiensis is a dynamic process involving germination, cell divisions, and sporulation in specific microhabitats. Recenti Prog Med, 2002 Apr, 93(4), 247 - 8 {Calmette-Guerin bacillus disseminated infection after intravesical instillation}; Borre S et al.; Local immunotherapy with bacillus Calmette-Guerin (BCG) is an effective and frequently used treatment for superficial bladder cancer . Serious side effects are infrequent but can affect every organ system . We describe a 53 year- old man with a disseminated bacillus Calmette-Guerin (BCG) infection after intravescical instillation for bladder carcinoma . Recent literature is reviewed for this rare complication. Microbiology, 2002 May, 148(Pt 5), 1571 - 9 Use of an arrayed promoter-probe library for the identification of macrophage-regulated genes in Mycobacterium tuberculosis; Hobson RJ et al.; The survival of Mycobacterium tuberculosis within the human host after infection, especially within macrophages, is likely to require the activation of a number of mycobacterial genes . To identify such genes, a promoter-probe library was constructed in which fragments of M . tuberculosis H37Rv DNA were inserted upstream of a lacZ reporter gene, using an Escherichia coli-mycobacterial shuttle vector . Mycobacterium bovis Bacille Calmette-Guerin (BCG) was subsequently transformed with this library and 4800 BCG clones were arrayed in a 96-well microtitre format, enabling the testing of individual clones for promoter activity under a variety of conditions . From preliminary screening, 41 clones were selected that exhibited upregulation of lacZ expression when subjected to acidified sodium nitrite . Subsequent sequence analyses identified 26 of these clones as containing potential promoters . After measuring lacZ expression in BCG clones recovered from a THP-1 macrophage cell line, three genes were selected for assessment of their expression in M . tuberculosis during macrophage infection, by real-time RT-PCR . Two of these genes, Rv1265 (with unknown function) and Rv2711 (encoding the iron-dependent repressor protein IdeR), showed evidence of being upregulated within macrophages. Biochim Biophys Acta, 2002 May 3, 1562(1-2), 63 - 9 Pore formation activity of Cry1Ab toxin from Bacillus thuringiensis in an improved membrane vesicle preparation from Manduca sexta midgut cell microvilli; Bravo A et al.; The pore formation activity of Cry1Ab toxin is analyzed in an improved membrane preparation from apical microvilli structures of Manduca sexta midgut epithelium cells (MEC) . A novel methodology is described to isolate MEC and brush border membrane vesicles (BBMV) from purified microvilli structures . The specific enrichment of apical membrane enzyme markers aminopeptidase (APN) and alkaline phosphatase (APh) were 35- and 22-fold, respectively, as compared to the whole midgut cell homogenate . Ligand-blot and Western-blot experiments showed that Cry1A specific receptors were also enriched . The pore formation activity of Cry1Ab toxin was fourfold higher in the microvilli membrane fraction that showed low intrinsic K+ channels and higher APN and APh activities than in the basal-lateral membrane fraction harboring high intrinsic K+ channels . These data suggest that basal-lateral membrane was separated from apical membrane.This procedure should allow more precise studies of the interaction of Cry toxins with their target membranes, avoiding unspecific interaction with other cellular membranes, as well as the study of the pore formation activity induced by Cry toxins in the absence of endogenous channels from M . sexta midgut cells. Huan Jing Ke Xue, 2002 Jan 30, 23(1), 74 - 7 {Water quality evaluation of the Three Gorges Reservoir area}; Liu H et al.; One year water quality monitoring results of 16 sections alongside the Three Gorges Reservoir area were presented . From the evaluation carried out using the integrated polluted index (P value), some characteristics of the water quality were obtained . The results showed that water quality of the Three Gorges Reservoir area was slightly polluted in 1999 mainly because of the discharge of urban sewerage and industrial wastewater . Main polluted city sections were along the river banks of Chongqing, Changshou, Fuling and Wanzhou . Evaluation also showed that there were 4 indicators that, to different extent, affected the water quality: colon bacillus, TP, non-ionic ammonia and oils. J Ind Microbiol Biotechnol, 2002 May, 28(5), 284 - 90 Physiological and molecular detection of crystalliferous Bacillus thuringiensis strains from habitats in the South Central United States; Ejiofor AO et al.; Gram-positive, endospore-forming Bacillus thuringiensis-like strains were isolated from 95 of 413 samples collected at the 0-5 cm depth of noncultivated soils and stagnant or dried-up ponds as well as from dust from stored grain products in South Central United States . Based on the production of parasporal crystals, 25 isolates were identified as B . thuringiensis after examining 227 B . thuringiensis-like colonies . The greatest proportion of samples yielding B . thuringiensis were from the dust from grain storage . The sodium acetate selective medium, heat processing, and crystal staining used in the initial screening revealed diverse populations of B . thuringiensis, which were categorized into distinct crystal morphological groups . Sugar fermentation, antibiotic sensitivity, growth characteristics and PCR studies showed diversity among the isolates that were distributed among 25 of the 58 known strains . The most frequently isolated strains were kurstaki, aizawai, morrisoni, thuringiensis, sotto and kenyae that together represented more than 90% of the characterized isolates . PCR analysis using 30 family primer pairs for cry and cyt genes showed that the frequency of the cry1 gene (62%) was predominant followed by the cry2 genes (30%), and the rest (8%) were other cry gene types, such as cry3, cry4, cry10, cry11, cry14, cry15, cry20, cry24 and cry26 . Both cyt1 and -2 genes were also detected . Several isolates showed PCR products on the gel that were not consistent with the expected sizes of nucleotides targeted by the primers . These were suggestive of nonspecific amplifications and were not used in the characterization process. J Ind Microbiol Biotechnol, 2002 Apr, 28(4), 193 - 200 Purification and characterization of maltooligosaccharide-forming amylase from Bacillus circulans GRS 313; Dey G et al.; A maltooligosaccharide-forming amylase that hydrolyzes starch into maltotriose and maltopentaose was found in the culture filtrate of a strain of Bacillus circulans GRS 313 isolated from local soil . The enzyme was purified by organic solvent fractionation, Sephadex G-100 gel filtration and CM-Sephadex column chromatography . Optimum pH and temperature of amylase were evaluated using response surface methodology (RSM) and were found to be 48 degrees C and 4.9, respectively . The enzyme was stable up to 60 degrees C and its pH stability was in the range of 5.0-8.0 . The Km and Vmax of the amylase with starch were 11.66 mg/ml and 68.97 U, respectively, and the energy of activation, Ea, was 7.52 kcal/mol . Dextrin inhibited the enzyme competitively, with a Ki of 6.1 mg/ml, and glucose caused noncompetitive inhibition with a Ki of 9.5 mg/ml . The enzyme was inhibited by Hg2+, Mn2+, Fe3+ and Cu2+ and enhanced by Co2+ and Mg2+ . EDTA reversed the inhibitory effect of the metals . Paper chromatographic and high-performance liquid chromatography analysis of the products of the amylolytic reaction showed the presence of maltotriose, maltotetraose, maltopentaose, maltose and glucose in the starch hydrolysate. Clin Exp Immunol, 2002 May, 128(2), 229 - 37 Immunological and pathological comparative analysis between experimental latent tuberculous infection and progressive pulmonary tuberculosis; Arriaga AK et al.; Mycobacterium tuberculosis produces latent infection or progressive disease . Indeed, latent infection is more common since it occurs in one-third of the world's population . We showed previously, using human material with latent tuberculosis, that mycobacterial DNA can be detected by in situ PCR in a variety of cell types in histologically-normal lung . We therefore sought to establish an experimental model in which this phenomenon could be studied in detail . We report here the establishment of such a model in C57Bl/6 x DBA/2 F1 hybrid mice by the intratracheal injection of low numbers of virulent mycobacteria (4000) . Latent infection was characterized by low and stable bacillary counts without death of animals . Histological and immunological study showed granulomas and small patches of alveolitis, with high expression of tumour necrosis factor alpha (TNFalpha), inducible nitiric oxide synthase (iNOS), interleukin 2 (IL-2) and interferon gamma (IFNgamma) . In contrast, the intratracheal instillation of high numbers of bacteria (1 x 106) produced progressive disease . These animals started to die after 2 months of infection, with very high bacillary loads, massive pneumonia, falling expression of TNF-alpha and iNOS, and a mixed Th1/Th2 cytokine pattern . In situ PCR to detect mycobacterial DNA revealed that the most common positive cells in latently-infected mice were alveolar and interstitial macrophages located in tuberculous lesions, but, as in latently-infected human lung, positive signals were also seen in bronchial epithelium, endothelial cells and fibroblasts from histologically-normal areas . Our results suggest that latent tuberculosis is induced and maintained by a type 1 cytokine pattern plus TNFalpha, and that mycobacteria persist intracellularly in lung tissue with and without histological evidence of a local immune response. Proc Natl Acad Sci U S A, 2002 Apr 30, 99(9), 5830 - 5 Loss of the membrane anchor of the target receptor is a mechanism of bioinsecticide resistance; Darboux I et al.; The mosquitocidal activity of Bacillus sphaericus is because of a binary toxin (Bin), which binds to Culex pipiens maltase 1 (Cpm1), an alpha-glucosidase present in the midgut of Culex pipiens larvae . In this work, we studied the molecular basis of the resistance to Bin developed by a strain (GEO) of C . pipiens . Immunohistochemical and in situ hybridization experiments showed that Cpm1 was undetectable in the midgut of GEO larvae, although the gene was correctly transcribed . The sequence of the cpm1(GEO) cDNA differs from the sequence we previously reported for a susceptible strain (cpm1(IP)) by seven mutations: six missense mutations and a mutation leading to the premature termination of translation . When produced in insect cells, Cpm1(IP) was attached to the membrane by a glycosylphosphatidylinositol (GPI) . In contrast, the premature termination of translation of Cpm1(GEO) resulted in the targeting of the protein to the extracellular compartment because of truncation of the GPI-anchoring site . The interaction between Bin and Cpm1(GEO) and the enzyme activity of the receptor were not affected . Thus, Bin is not toxic to GEO larvae because it cannot interact with the midgut cell membrane, even though its receptor site is unaffected . This mechanism contrasts with other known resistance mechanisms in which point mutations decrease the affinity of binding between the receptor and the toxin. Clin Exp Immunol, 2002 Apr, 128(1), 52 - 8 Prostaglandin E2 down-regulates viable Bacille Calmette-Guérin-induced macrophage cytotoxicity against murine bladder cancer cell MBT-2 in vitro; Yamada H et al.; The regulatory effect of prostaglandin (PG) E2 and a cyclooxygenase (COX) inhibitor on Bacille Calmette-Guerin (BCG)-induced macrophage cytotoxicity in a bladder cancer cell, MBT-2, was studied in vitro . BCG stimulated thioglycollate-elicited murine peritoneal exudate cells (PEC) to induce cytotoxic activity and to produce cytokines such as interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and PGE2 . NS398, a specific COX-2 inhibitor, and indomethacin (IM), a COX-1 and COX-2 inhibitor, enhanced viable BCG-induced cytotoxic activity and IFN-gamma and TNF-alpha production of PEC . However, NS398 and IM did not enhance these activities induced by killed BCG . Enhanced cytotoxicity was mediated by increased amounts of IFN-gamma and TNF-alpha . Exogenous PGE2 reduced cytotoxic activity and IFN-gamma and TNF-alpha production of PEC . These results suggest that PGE2 produced by BCG-activated macrophages has a negative regulatory effect on the cytotoxic activity of macrophages . Accordingly, a PG synthesis inhibitor may be a useful agent to enhance BCG-induced antitumour activity of macrophages. J Agric Food Chem, 2002 May 8, 50(10), 2812 - 7 Cooperative action of alpha-glucanotransferase and maltogenic amylase for an improved process of isomaltooligosaccharide (IMO) production; Lee HS et al.; Maltogenic amylase and alpha-glucanotransferase (alpha-GTase) were employed in an effort to develop an efficient process for the production of isomaltooligosaccharides (IMOs) . Bacillus stearothermophilus maltogenic amylase (BSMA) and alpha-GTase from Thermotoga maritima were overexpressed in Escherichia coli using overexpression vectors . An IMO mixture containing 58% of various IMOs was produced from liquefied corn syrup by the hydrolyzing and transglycosylation activities of BSMA alone . When BSMA and alpha-GTase were reacted simultaneously, the IMO content increased to 68% and contained relatively larger IMOs compared with the products obtained by the reaction without alpha-GTase . Time course analysis of the IMO production suggested that BSMA hydrolyzed maltopentaose and maltohexaose most favorably into maltose and maltotriose and transferred the resulting molecules simultaneously to acceptor molecules to form IMOs . alpha-GTase transferred donor sugar molecules to the hydrolysis products such as maltose and maltotriose to form maltopentaose, which was then rehydrolyzed by BSMA as a favorable substrate. Redox Rep, 2002, 7(1), 29 - 33 Phospholipase C-dependent hydrolysis of phosphatidylcholine hydroperoxides to diacylglycerol hydroperoxides and its reduction by phospholipid hydroperoxide glutathione peroxidase; Kambayashi Y et al.; We have shown that 1,2-diacylglycerol hydroperoxides activate protein kinase C (PKC) as efficiently as does phorbol ester {Takekoshi S, Kambayashi Y, Nagata H, Takagi T, Yamamoto Y, Watanabe K . Activation of protein kinase C by oxidized diacylglycerol . Biochem Biophys Res Commun 1995; 217: 654-660} . 1,2-Diacylglycerol hydroperoxides also stimulate human neutrophils to release superoxide whereas their hydroxides do not {Yamamoto Y, Kambayashi Y, Ito T, Watanabe K, Nakano M . 1,2-Diacylglycerol hydroperoxides induce the generation and release of superoxide anion from human polymorphonuclear leukocytes . FEBS Lett 1997; 412: 461-464} . One of the proposed mechanisms for the formation of 1,2-diacylglycerol hydroperoxides is the hydrolysis of phosphatidylcholine hydroperoxides by phospholipase C (PLC) . To confirm this hypothesis, we incubated 1-palmitoyl-2-linoleoyl-phosphatidylcholine (PLPC) liposomes containing PLPC hydroperoxides (PLPC-OOH) with Bacillus cereus PLC and found 1-palmitoyl-2-linoleoylglycerol (PLG) and its hydroperoxide (PLG-OOH) were produced . PLC hydrolyzed the two substrates without preference, as the yields of PLG and PLG-OOH were the same even though cholesterol was incorporated into liposomes to increase bilayer integrity . Phospholipid hydroperoxide glutathione peroxidase (PHGPX) reduced PLG-OOH to its hydroxide in the presence of glutathione while the conventional cytosolic glutathione peroxidase did not . These data suggest that PLC hydrolyzes oxidized biomembranes to give 1,2-diacylglycerol hydroperoxides for PKC stimulation but PHGPX may prevent neutrophil stimulation by reducing 1,2-diacylglycerol hydroperoxides to their hydroxides. Braz J Infect Dis, 2002 Feb, 6(1), 40 - 4 Epub 2003 Mar 18. Reiter's syndrome associated with the Acquired Immunodeficiency Syndrome: a case report; Malta JB et al.; The association of Reiter's Syndrome (RS) with the Acquired Immunodeficiency Syndrome (AIDS) is seldom mentioned in the medical literature . This report illustrates this relationship in a 46 years old male patient suffering from AIDS (CD(4)(+) = 240 cells/mm(3), CD(8)(+) = 1,301 cells/mm(3) and viral load = 330,000 copies/ml), pulmonary tuberculosis (positive catarrhal bacilluscopy), and RS . The diagnosis of RS was based on the combination of dermatological and articular alterations . The patient s cutaneous lesions were characterized by exfoliation and the formation of crusts located on the face, scalp, genitals, hands, and feet; onychodystrophy with opacity; yellowish coloring; and hyperkeratosis of the nails . Articular lesions led to progressive deformity of phalangeal joints of the hands, and intensive arthralgia, mainly of the larger joints (shoulders, elbows, hips and knees) . AIDS treatment was administered with anti-retroviral drugs (zidovudine and didanosine); for tuberculosis (isoniazid, rifampicine, and pyrazinamide); and (prednisone and inometacine) for the RS . The patient recovered with the improvement of articular symptoms; however, on the eighth day of treatment, the patient showed significant hemoptysis and hypovolemic shock, and died . The association of RS and HIV infection is reviewed. Acta Crystallogr D Biol Crystallogr, 2002 May, 58(Pt 5), 744 - 50 Epub 2002 Apr 26. The structure of substrate-free microbial ribonuclease binase and of its complexes with 3'GMP and sulfate ions; Polyakov KM et al.; The structures of Bacillus intermedius ribonuclease (binase), an extracellular 109-residue enzyme, and its complexes with 3'GMP and sulfate ions were solved at 1.65 and 2.0 A, respectively . The structures were refined using REFMAC . The crystal of free binase belongs to the space group C2, whereas the crystals of complexes belong to the space group P2(1)2(1)2(1) . In both crystal lattices the asymmetric unit contains two molecules which form an identical dimer . The structure of the dimer is such that only one of its subunits can bind the nucleotide in the 3'GMP-binase complex, where the guanyl base is located in the recognition loop of the enzyme . In both binase complex structures the phosphate group of 3'GMP or one of the sulfate ions make an electrostatic interaction with the binase molecule at the catalytic site . A second phosphate-binding site was found in the structures of the complexes at the cleft formed by the loop 34-39, the main chain of Arg82 and the side chain of Trp34 . Comparison of the complex and unliganded enzyme crystal structures shows that there are some small but distinct differences in the specificity loop (56-62) and in the loops 34-39 and 99-104 associated with the binding of the nucleotide and sulfate ions. Appl Environ Microbiol, 2002 May, 68(5), 2479 - 83 Quantitative analysis of cereulide, the emetic toxin of Bacillus cereus, produced under various conditions; Haggblom MM et al.; This paper describes a quantitative and sensitive chemical assay for cereulide, the heat-stable emetic toxin produced by Bacillus cereus . The methods previously available for measuring cereulide are bioassays that give a toxicity titer, but not an accurate concentration . The dose of cereulide causing illness in humans is therefore not known, and thus safety limits for cereulide cannot be indicated . We developed a quantitative and sensitive chemical assay for cereulide based on high-performance liquid chromatography (HPLC) connected to ion trap mass spectrometry . This chemical assay and a bioassay based on boar sperm motility inhibition were calibrated with purified cereulide and with valinomycin, a structurally similar cyclic depsipeptide . The boar spermatozoan motility assay and chemical assay gave uniform results over a wide range of cereulide concentrations, ranging from 0.02 to 230 microg ml(-1) . The detection limit for cereulide and valinomycin by HPLC-mass spectrometry was 10 pg per injection . The combined chemical and biological assays were used to define conditions and concentrations of cereulide formation by B . cereus strains F4810/72, NC7401, and F5881 . Cereulide production commenced at the end of logarithmic growth, but was independent of sporulation . Production of cereulide was enhanced by incubation with shaking compared to static conditions . The three emetic B . cereus strains accumulated 80 to 166 microg of cereulide g(-1) (wet weight) when grown on solid medium . Strain NC7401 accumulated up to 25 microg of cereulide ml(-1) in liquid medium at room temperature (21 +/- 1 degrees C) in 1 to 3 days, during the stationary growth phase when cell density was 2 x 10(8) to 6 x 10(8) CFU ml(-1) . Cereulide production at temperatures at and below 8 degrees C or at 40 degrees C was minimal. Appl Environ Microbiol, 2002 May, 68(5), 2106 - 12 Denaturation of either Manduca sexta aminopeptidase N or Bacillus thuringiensis Cry1A toxins exposes binding epitopes hidden under nondenaturing conditions; Daniel A et al.; The effect of polypeptide denaturation of Bacillus thuringiensis Cry1A toxins or purified Manduca sexta 120-kDa aminopeptidase N on the specificities of their interactions was investigated . Ligand and dot blotting experiments were conducted with (125)I-labeled Cry1Ac, Cry1Ac mutant (509)QNR-AAA(511) (QNR-AAA), or 120-kDa aminopeptidase N as the probe . Mutant QNR-AAA does not bind the N-acetylgalactosamine moiety on the 120-kDa aminopeptidase . Both (125)I-Cry1Ac and (125)I-QNR-AAA bound to 210- and 120-kDa proteins from M . sexta brush border membrane vesicles and purified 120-kDa aminopeptidase N on ligand blots . However, on dot blots (125)I-QNR-AAA bound brush border vesicles but did not bind purified aminopeptidase except when aminopeptidase was denatured . In the reciprocal experiment, (125)I-aminopeptidase bound Cry1Ac but did not bind QNR-AAA . (125)I-aminopeptidase bound Cry1Ab to a limited extent but not the Cry1Ab domain I mutant Y153D or Cry1Ca . However, denatured (125)I-aminopeptidase detected each Cry1A toxin and mutant but not Cry1Ca on dot blots . The same pattern of recognition occurred with native (nondenatured) (125)I-aminopeptidase probe and denatured toxins as the targets . The broader pattern of toxin-binding protein interaction is probably due to peptide sequences being exposed upon denaturation . Putative Cry toxin-binding proteins identified by the ligand blot technique need to be investigated under native conditions early in the process of identifying binding proteins that may serve as functional toxin receptors. Life Sci Space Res, 1967, 5, 174 - 86 Effect of reduced barometric pressure on water availability related to microbial growth; Hawrylewicz EJ et al.; Data obtained from Mariner IV indicate that the barometric pressure on Mars is considerably lower than previously estimated . Current estimates from Mariner IV indicate a range from 4 to 7 mb and by near infrared spectroscopy 33-56 mb . Inasmuch as the pressure has a marked influence on availability of water, this should affect the existence of Martian life . At the maximum temperatures recorded on Mars, namely 25 degrees C, a barometric pressure of 30 mb is required for the retention of free water . The lower pressure, 4 mb, would suggest that the moisture is present as a vapor above the freezing point and consequently it is not available for utilizing by living cells . The lower estimates of barometric pressure also inversely affect the carbon dioxide concentration in the Martian atmosphere . Our previous studies have demonstrated that spores of Bacillus cereus survive, germinate and grow in a simulated Martian environment (2.4% CO2, 98 mb) supplemented with moisture . The studies described in this paper were designed to determine the effect of low barometric pressures (10 to 98 mb Hg) and high concentrations of carbon dioxide (37 to 100%) in the simulated Martian environment on survival and growth of B cereus . The organism was inoculated into a felsite-limonite soil at 8% moisture level . The temperature cycles used were 8 hr at -65 degrees C and 16 hr at 25 degrees C, or 20 hr at -65 degrees C and 4 hr at 25 degrees C . The data suggest that the organism after achieving maximum growth in the simulated Martian environment (2.4% CO2, 98 mb) immediately enters into the growth phase upon reinoculation into fresh soil . These data reflect upon the possibility of contamination through air movements . Based upon currently available Martian environmental data, the probability of contamination of Mars by terrestrial micro-organisms will be discussed. J Appl Microbiol, 2002, 92(5), 983 - 91 Polymerase chain reaction identification of Bacillus sporothermodurans from dairy sources; Scheldeman P et al.; AIMS: A new polymerase chain reaction (PCR) method for the identification of Bacillus sporothermodurans strains from sterilized or ultrahigh temperature-treated milk and milk products and from other non-milk sources and environments, including the dairy farm . METHODS AND RESULTS: Two strains from raw milk and feed concentrate could be allocated to B . sporothermodurans based on 16S rDNA sequencing and DNA-DNA hybridization results . Two specific PCR primers were derived from the 16S rRNA gene of B . sporothermodurans . CONCLUSIONS: The PCR identification method was validated using a collection of B . sporothermodurans strains from different sources and on a large collection of dairy and non-dairy Bacillus spp . and other relevant taxa . SIGNIFICANCE AND IMPACT OF THE STUDY: This PCR method was used as a screening method for strains with very heat-resistant endospores, isolated at the dairy farm level after heat treatment for 30 min at 100 degrees C . Seventeen strains isolated at the dairy farm were identified as B . sporothermodurans . They originated mainly from feed concentrate and also from soy, pulp and silage . The PCR identification method described here can, therefore, contribute to a better understanding of the route by which B . sporothermodurans contaminates raw and/or heat-treated milk. J Appl Microbiol, 2002, 92(5), 912 - 9 Discrimination of Bacillus cereus and Bacillus thuringiensis with 16S rRNA and gyrB gene based PCR primers and sequencing of their annealing sites; Chen ML et al.; AIMS: To evaluate the possibility for discrimination of Bacillus cereus and B . thuringiensis using 16S rRNA and gyrB gene based PCR methods, and to obtain the sequences of the primer annealing sites so that the PCR results may be explained . METHODS AND RESULTS: Based on the sequence difference in the variable region (V1) of 16S rRNA and in the gyrB gene between B . cereus and B . thuringiensis, PCR primers specific to these Bacillus spp . were designed . When these primers were used to discriminate B . cereus and B . thuringiensis, six of 82 B . cereus strains were identified as B . thuringiensis while 67 of 73 B . thuringiensis strains were identified as B . cereus . Sequence analysis of the primer annealing sites showed that there is no clear-cut difference in the V1 region of 16S rRNA, and in the gyrB gene, between B . cereus and B . thuringiensis strains . CONCLUSIONS: Although 16S rDNA based probes and gyrB gene based PCR primers have been suggested for the discrimination of B . cereus and B . thuringiensis strains, when a large number of Bacillus strains was tested, results showed that discrimination between B . cereus and B . thuringiensis is difficult . Therefore, to distinguish B . thuringiensis from B . cereus, a single feature, such as the presence of a parasporal crystal protein or cry gene, may sometimes be reliable . SIGNIFICANCE AND IMPACT OF THE STUDY: Discrimination between B . cereus and B . thuringiensis is a challenging debate to which this paper makes a contribution. J Appl Microbiol, 2002, 92(5), 879 - 84 Bacillus megaterium spore germination is influenced by inoculum size; Caipo ML et al.; AIMS: The effect of spore density on the germination (time-to-germination, percent germination) of Bacillus megaterium spores on tryptic soy agar was determined using direct microscopic observation . METHODS AND RESULTS: Inoculum size varied from approximately 10(3) to 10(8) cfu ml(-1) in a medium where pH=7 and the sodium chloride concentration was 0.5% w/v . Inoculum size was measured by global inoculum size (the concentration of spores on a microscope slide) and local inoculum size (the number of spores observed in a given microscope field of observation) . Both global and local inoculum sizes had a significant effect on time-to-germination (TTG), but only the global inoculum size influenced the percentage germination of the observed spores . CONCLUSIONS: These results show that higher concentrations of Bacillus megaterium spores encourage more rapid germination and more spores to germinate, indicating that low spore populations do not behave similarly to high spore populations . SIGNIFICANCE AND IMPACT OF THE STUDY: A likely explanation for the inoculum size-dependency of germination would be chemical signalling or quorum sensing between Bacillus spores. Int J Urol, 2002 Jan, 9(1), 29 - 35 Surface antigen expression on bladder tumor cells induced by bacillus Calmette-Guérin (BCG): A role of BCG internalization into tumor cells; Ikeda N et al.; BACKGROUND: The antitumor mechanisms of bacillus Calmette-Guerin (BCG) against bladder cancer is still unclear . We previously reported that BCG was internalized by and survived within murine bladder tumor cells (MBT-2) for at least 40 days . In the present study, we investigated the effect of BCG on the surface antigen expression of bladder tumor cells and the characteristics of these cells as antigen-presenting cells in vitro . METHODS: Surface antigen (major histocompatibility complex (MHC) Class II, CD1, CD80 and intercellular adhesion molecule-1 (ICAM-1)) expression on BCG-treated murine (MBT-2) and human (T-24, J82) bladder tumor cells were analyzed using flow cytometry . The production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) from murine lymphocytes sensitized with BCG or BCG-treated tumor cells were also investigated . RESULTS: The expressions of MHC Class II, CD1, CD80 and ICAM-1 were augmented in all of the bladder tumor cell lines used; however, they were augmented to varying degrees among the cell lines that were treated with live BCG . Heat-killed BCG had little or no effect . When murine lymph node cells sensitized with BCG or BCG-treated MBT-2 cells were cocultured with BCG-treated MBT-2 cells, significant amounts of IL-2 and IFN-gamma were produced in the culture medium . CONCLUSIONS: BCG induced the augmented expression of surface antigens, such as MHC Class II, CD1, CD80 and ICAM-1, of bladder tumor cells . Furthermore, BCG-treated MBT-2 cells could stimulate BCG-sensitized lymphocytes to produce IL-2 and IFN-gamma . These results strongly suggest that bladder tumor cells gained the characteristics and functions of antigen-presenting cells (APC). Ecotoxicol Environ Saf, 2002 Mar, 51(3), 177 - 86 Effect of insecticides on plankton and fish of Ignacio Ramirez reservoir (Mexico): a biochemical and biomagnification study; Favari L et al.; Studies on the limnology, plankton, and biomagnification of pesticides at Ignacio Ramirez (IR) reservoir (Mexico) were undertaken . The reservoir is located in central Mexico, in an agricultural zone with high soil erosion . Acetylcholinesterase (AchE), gamma-glutamyl transpeptidase (GGTP), and lipid peroxidation were assayed in fish . Organochlorines (0.024-0.279 mg/liter) and organophosphates (0.02 x 10(-3)-0.21 x 10(-3) mg/liter) were present at high concentrations in water and the biota assayed . In the IR dam the plankton fluctuated depending on the dry and wet seasons . The dominant group of phytoplankton was Bacillariophyta (20-85%) in May, Cyanophyta (22-65%) in September, and Cyanophycean (10-65%), Chlorophycean (10-60%), and Bacyllariophycean (5-80%) species in March . The zooplankton were dominated by cladoceran species (40-70%) . Organochlorine and organophosphate insecticides were bioconcentrated (2- to 10-fold) from water to algae, 10- to 25-fold in zooplankton, and 8- to 140-fold in fish . GGTP activity and lipid peroxidation increased and AchE activity in fish decreased in response to the environmental stress caused by the elevated biomagnification of pesticides . The bioaccumulation of these contaminants in fish and the potential for biomagnification in humans are perceived as threats . (c) 2002 Elsevier Science (USA). Plant Cell, 2002 Apr, 14(4), 795 - 803 Rice TATA binding protein interacts functionally with transcription factor IIB and the RF2a bZIP transcriptional activator in an enhanced plant in vitro transcription system; Zhu Q et al.; TATA binding protein (TBP) and transcription factor IIB (TFIIB) are key factors for the assembly of eukaryotic transcription initiation complexes . We used a rice whole-cell extract in vitro transcription system to characterize the functional interactions of recombinant plant TBP and TFIIB . Bacterially expressed rice TBP (OsTBP2) bound to the TATA box of the rice pal gene encoding phenylalanine ammonia-lyase, caused DNA bending, and enhanced basal transcription from the pal promoter in a TATA box-dependent manner . Recombinant rice TFIIB (OsTFIIB) stimulated the DNA binding and bending activities of OsTBP2 and synergistically enhanced OsTBP2-mediated transcription from the pal promoter and the promoter of Rice tungro bacilliform virus but not from the barley pr1 promoter . We also demonstrate a physical interaction between OsTBP2 and RF2a, a rice bZIP transcription factor that bound to the box II cis element of the promoter of Rice tungro bacilliform virus, resulting in enhanced transcription from the viral promoter . Enhancement of rice whole-cell extracts with recombinant transcription factors thus provides a powerful tool for the in vitro determination of plant gene regulation mechanisms . We conclude that OsTBP2 undergoes promoter-specific functional interactions with both the basal transcription factor OsTFIIB and the accessory transcription factor RF2a. Ann Urol (Paris), 2002 Mar, 36(2), 120 - 31 {Classification, favorable characteristics, prevention and treatment of adverse side-effects associated with Bacillus Calmette-Guerin in the treatment of superficial bladder cancer}; Saint F et al.; The efficacy of Bacillus Calmette-Guerin (BCG) in the treatment of superficial bladder cancer was first reported by Morales in 1976 . Several authors have since demonstrated the efficacy of BCG in the prophylaxis and treatment of high-risk superficial bladder tumors (pT1G3, CIS) . Although BCG is now recommended as an adjunctive treatment for superficial bladder tumors, the optimal treatment schedule remains to be defined . Results reported by Lamm suggest that an initial induction cycle of six weekly intravesical BCG instillations is suboptimal unless maintenance therapy (three consecutive weekly instillations) is given 3, 6, 12, 18, 24, 30 and 36 months later . However, the use of maintenance therapy is hindered by troublesome adverse reactions . This article reviews adverse reactions associated with BCG treatment, proposed a classification and discusses their prevention and treatment. Protein Sci, 2002 May, 11(5), 1091 - 100 Thermodynamic analysis of the binding of component enzymes in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus; Jung HI et al.; The peripheral subunit-binding domain (PSBD) of the dihydrolipoyl acetyltransferase (E2, EC 2.3.1.12) binds tightly but mutually exclusively to dihydrolipoyl dehydrogenase (E3, EC 1.8.1.4) and pyruvate decarboxylase (E1, EC 1.2.4.1) in the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus . Isothermal titration calorimetry (ITC) experiments demonstrated that the enthalpies of binding (DeltaH degrees ) of both E3 and E1 with the PSBD varied with salt concentration, temperature, pH, and buffer composition . There is little significant difference in the free energies of binding (DeltaG degrees = -12.6 kcal/mol for E3 and = -12.9 kcal/mol for E1 at pH 7.4 and 25 degrees C) . However, the association with E3 was characterized by a small, unfavorable enthalpy change (DeltaH degrees = +2.2 kcal/mol) and a large, positive entropy change (TDeltaS degrees = +14.8 kcal/mol), whereas that with E1 was accompanied by a favorable enthalpy change (DeltaH degrees = -8.4 kcal/mol) and a less positive entropy change (TDeltaS degrees = +4.5 kcal/mol) . Values of DeltaC(p) of -316 cal/molK and -470 cal/molK were obtained for the binding of E3 and E1, respectively . The value for E3 was not compatible with the DeltaC(p) calculated from the nonpolar surface area buried in the crystal structure of the E3-PSBD complex . In this instance, a large negative DeltaC(p) is not indicative of a classical hydrophobic interaction . In differential scanning calorimetry experiments, the midpoint melting temperature (T(m)) of E3 increased from 91 degrees C to 97.1 degrees C when it was bound to PSBD, and that of E1 increased from 65.2 degrees C to 70.0 degrees C . These high T(m) values eliminate unfolding as a major source of the anomalous DeltaC(p) effects at the temperatures (10-37 degrees C) used for the ITC experiments. J Biol Chem, 2002 Jul 5, 277(27), 24142 - 7 Epub 2002 Apr 19. Substrate-activated zinc binding of metallo-beta -lactamases: physiological importance of mononuclear enzymes; Wommer S et al.; We have investigated the influence of substrate binding on the zinc ion affinity of representatives from the three metallo-beta-lactamase subclasses, B1 (BcII from Bacillus cereus and BlaB from Chryseobacterium meningosepticum), B2 (CphA from Aeromonas hydrophila), and B3 (L1 from Stenotrophomonas maltophilia) . By competition experiments with metal-free apoenzymes and chromophoric zinc chelators or EDTA, we determined the dissociation constants in the absence and presence of substrates . For the formation of the monozinc enzymes we determined constants of 1.8, 5.1, 0.007, and 2.6 nm in the absence and 13.6, 1.8, 1.2, and 5.7 pm in the presence of substrates for BcII, BlaB, CphA, and L1, respectively . A second zinc ion binds in the absence (presence) of substrates with considerably higher dissociation constants, namely 1.8 (0.8), 0.007 (0.025), 50 (1.9), and 0.006 (0.12) microm for BcII, BlaB, CphA, and L1, respectively . We have concluded that the apo form might be the prevailing state of most of the metallo-beta-lactamases under physiological conditions in the absence of substrates . Substrate availability induces a spontaneous self-activation due to a drastic decrease of the dissociation constants, resulting in the formation of active mononuclear enzymes already at picomolar free zinc ion concentrations . In the presence of substrates, the binuclear state of the enzymes only exists at unphysiologic high zinc concentrations and might be of no biological relevance . From the competition experiments with EDTA it is further concluded that the reactivation rate does not depend on the pool of free zinc ions but proceeds via the EDTA-Zn(II)-enzyme ternary complexes. Lett Appl Microbiol, 2002, 34(5), 317 - 22 Improved polygalacturonase production from Bacillus sp . MG-cp-2 under submerged (SmF) and solid state (SSF) fermentation; Kapoor M et al.; AIMS: To investigate the effect of amino acids, vitamins and surfactants on polygalacturonase production from Bacillus sp . MG-cp-2 under submerged (SmF) and solid state fermentation (SSF) . METHODS AND RESULTS: Bacillus sp . MG-cp-2 was isolated from the outer covering of the seeds of Celastrus paniculatus . Out of the various surfactants, amino acids and vitamins, Tween-60, DL-serine and folic acid maximally enhanced polygalacturonase production by 2.7-fold (240.0 U x ml(-1)), 4.0-fold (360.0 U x ml(-1)) and 3.8-fold (342.0 U x ml(-1)) respectively, under submerged fermentation (SmF) . In solid state fermentation (SSF), Tween-80, pyridoxine and DL-ornithine monohydrochloride induced highest enzyme production up to 1.73-fold (6956.5 U x g(-1)), 5.3-fold (21224.4 U x g(-1)) and 5.74-fold (23076.9 U x g(-1)), respectively . CONCLUSION: Amino acids and their analogues, vitamins and surfactants effect significantly polygalacturonase production by Bacillus sp . MG-cp-2 when grown under submerged (SmF) and solid state fermentation (SSF) conditions . SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides useful information about regulation of polygalacturonase biosynthesis in Bacillus sp . MG-cp-2, which appears to be an interplay of nutritional and physical factors . Alkaline polygalacturonase from Bacillus sp . MG-cp-2 will be extremely useful in the treatment of alkaline pectic waste waters from vegetable and fruit processing industries and in degumming of bast fibres. J Appl Microbiol, 2002, 92(4), 776 - 83 The influence of nutritional and environmental conditions on the accumulation of poly-beta-hydroxybutyrate in Bacillus mycoides RLJ B-017; Borah B et al.; AIMS: To optimize the nutritional and environmental conditions for growth of and poly-beta-hydroxybutyrate (PHB) accumulation in Bacillus mycoides RLJ B-017 . METHODS AND RESULTS: An isolate, identified as B . mycoides, was grown on different sources of carbon and nitrogen . Among these, sucrose, beef extract and di-ammonium sulphate were found to be the most suitable for growth and PHB accumulation . The overall maximum value of PHB (%) in cells, PHB yield (Yp/s) and productivities (Qp and qp) were 69.4 +/- 0.4% dry cell weight (DCW), 0.21 gp gS(-1), 0.104 +/- 0.012 gp l(-1) h(-1) and 0.03 gp gx(-1) h(-1), respectively when grown in a medium containing 20 gs l(-1) sucrose, supplemented with di-ammonium sulphate . The addition of beef extract increased the value of PHB (%) in cells, PHB yield and productivities by 17.58 +/- 0,3, 23.8, 19.23 +/- 0.3 and 13.8 +/- 0.2% , respectively . The overall maximum values of PHB (% DCW), PHB yield and productivities were obtained at pH 7.0 +/- 0 .1, temperature 30 +/- 0.5 degrees C, agitation 650 rev min(-1) and oxygen transfer rate 3.8 mmol O(2) l(-1) h(-1) . CONCLUSIONS: Sucrose, glucose and fructose were found to be more suitable for cell growth and PHB accumulation, but sucrose was less expensive than glucose . Among the nitrogen sources, beef extract and di-ammonium sulphate promoted PHB synthesis . The accumulation of PHB was observed to be growth associated . SIGNIFICANCE AND IMPACT OF THE STUDY: Gram-positive bacteria have not been reported to accumulate large amounts of polyhydroxyalkanoate and hence have not been considered as potent candidates for industrial production . A number of Bacillus spp . have been reported to accumulate 9-44.5% DCW PHB . By comparison, Bacillus RLJ B-017 contained 69.4 +/- 0.4% DCW PHB . Therefore, this strain has been considered as a potent organism for industrial interest . A relatively high yield of PHB was obtained in this wild strain and PHB synthesis was independent of nutrient limitation . The conditions for the higher PHB yield and productivity will be optimized in the next phase using fed-batch culture. J Appl Microbiol, 2002, 92(4), 745 - 52 Contents of cry genes and insecticidal toxicity of Bacillus thuringiensis strains from terrestrial and aquatic habitats; Martinez C et al.; AIMS: Two Bacillus thuringiensis collections from terrestrial and aquatic habitats were compared in order to study the possible interrelationships between habitat and biological characteristics (serovar, cry genes content and toxicity) . METHODS AND RESULTS: Bacillus thuringiensis strains were characterized by serology, PCR, and one-dose treatment against the noctuids Helicoverpa armigera and Spodoptera exigua, and the dipteran Tipula oleracea . A total of 12 and 10 different serovars were identified within terrestrial and aquatic strains, respectively . The number of non-toxic strains was greater in aquatic (41.6%) than in terrestrial habitats (5.3%) . The genes cry1C, cry1D and cry1E were significantly more frequent in the terrestrial habitat . The cry1B gene was very frequent within thuringiensis strains . CONCLUSIONS: A high diversity was found in terms of serovars present and cry genes content in both collections . The relative frequency of individual cry genes was different in both collections, and a serovar-dependent distribution of the cry1B gene was found . Some strains sharing the same set of cry genes differed in their toxicity, suggesting important differences in gene expression . SIGNIFICANCE AND IMPACT OF THE STUDY: The inter-relationships between serology, cry gene content and toxicity may allow a better understanding of B . thuringiensis ecology. BJU Int, 2002 May, 89(7), 671 - 80 Long-term follow-up of a randomized prospective trial comparing a standard 81 mg dose of intravesical bacille Calmette-Guérin with a reduced dose of 27 mg in superficial bladder cancer; Martinez-Pineiro JA et al.; OBJECTIVES: To determine the efficacy of a three-fold reduced dose (RD, 27 mg) of intravesical bacille Calmette-Guerin (BCG) against the standard dose (81 mg) in patients with superficial bladder cancer, assessing recurrence, progression and differences in toxicity . PATIENTS AND METHODS: Five hundred patients with superficial bladder cancer (Ta, T1, Tis) were enrolled and randomly assigned to be treated after transurethral resection of all visible lesions with intravesical BCG Connaught strain (weekly x six and thereafter fortnightly x six) either with the standard or RD instillation . RESULTS: All but one of the 500 patients were evaluable for efficacy and toxicity (252 in the standard arm and 247 in the RD arm) . The median follow-up was 69 months (maximum 104); 71 (28%) patients in the standard arm and 76 (31%) in the RD arm developed recurrences; the median time to recurrence has not yet been attained, but at 5 years the mean (sd) percentage of recurrence-free patients was 70.5 (3.12) and 70.4 (3.1) for the standard and RD arms, respectively . In patients presenting with multifocal tumours, the standard dose was more effective against recurrences than the RD (P=0.0151) . In those with G3 and high-risk tumours overall, the superiority of the standard dose was marginal (P=0.060 and P=0.082) . Twenty-nine (11.5%) tumours in the standard arm and 33 (13.3%) in the RD arm progressed to invasive disease; the median time to progression has not yet been attained, but the percentage of progression-free patients at 5 years was 88.8 (2.23) and 86.9 (2.31) for the standard and RD arms, respectively . The standard dose was more effective than the RD against progression only in patients with multifocal disease (P=0.048) . Twelve (4.8%) cystectomies were performed in the standard and 15 (6.1%) in the RD arm . Currently, 106 (21.2%) patients have died, but only 38 (7.6%) from bladder cancer, i.e . 20 (7.9%) in the standard and 18 (7.5%) in RD arm . Overall the disease-specific death rate was lower for those patients who completed the scheduled treatment . The cause-specific survival at 5 years did not differ between the arms (P=0.76) but there was a trend toward better cause-specific survival for patients with multifocal tumours in the standard arm . Toxicity differed between the arms, significantly more patients having no toxicity in the RD arm, and fewer having delayed instillations or withdrawing . However, severe systemic toxicity occurred even in patients treated with the RD, in a similar proportion to those receiving the standard dose . CONCLUSION: Overall, the RD gave similar results for recurrence and progression but with significantly less toxicity . However, patients with multifocal tumours fared better with the standard dose and there was a trend towards better recurrence rates in patients with high-risk tumours . We recommend continuing to use the standard dose for high-risk tumours, while we consider the reduced dose safe and effective for intermediate-risk lesions and for maintenance schedules. Curr Opin Allergy Clin Immunol, 2001 Dec, 1(6), 503 - 11 Mycobacterial diseases in primary immunodeficiencies; Reichenbach J et al.; Primary immunodeficiency diseases comprise over 100 conditions, each associated with a variety of viral, bacterial, fungal and protozoan infections . M . tuberculosis and less virulent mycobacteria, such as bacille Calmette-Guerin vaccines and environmental non-tuberculous mycobacteria, may cause severe disease in patients with primary immunodeficiency diseases . However, no previous review has dealt with the issue of which primary immunodeficiency diseases predispose affected individuals to mycobacterial disease . This information is very useful, not only increasing our understanding of human immunity to mycobacteria, but also for the diagnostic investigation of patients with mycobacteriosis . We review here the medical literature on cases of mycobacterial disease in patients with primary immunodeficiency diseases. Biochem J, 2002 May 1, 363(Pt 3), 687 - 96 Mutational analysis of the two zinc-binding sites of the Bacillus cereus 569/H/9 metallo-beta-lactamase; de Seny D et al.; The metallo-beta-lactamase BcII from Bacillus cereus 569/H/9 possesses a binuclear zinc centre . The mono-zinc form of the enzyme displays an appreciably high activity, although full efficiency is observed for the di-zinc enzyme . In an attempt to assign the involvement of the different zinc ligands in the catalytic properties of BcII, individual substitutions of selected amino acids were generated . With the exception of His(116)-->Ser (H116S), C221A and C221S, the mono- and di-zinc forms of all the other mutants were poorly active . The activity of H116S decreases by a factor of 10 when compared with the wild type . The catalytic efficiency of C221A and C221S was zinc-dependent . The mono-zinc forms of these mutants exhibited a low activity, whereas the catalytic efficiency of their respective di-zinc forms was comparable with that of the wild type . Surprisingly, the zinc contents of the mutants and the wild-type BcII were similar . These data suggest that the affinity of the beta-lactamase for the metal was not affected by the substitution of the ligand . The pH-dependence of the H196S catalytic efficiency indicates that the zinc ions participate in the hydrolysis of the beta-lactam ring by acting as a Lewis acid . The zinc ions activate the catalytic water molecule, but also polarize the carbonyl bond of the beta-lactam ring and stabilize the development of a negative charge on the carbonyl oxygen of the tetrahedral reaction intermediate . Our studies also demonstrate that Asn(233) is not directly involved in the interaction with the substrates. Biochem J, 2002 May 1, 363(Pt 3), 437 - 47 Characterization of a putative alpha-mannosyltransferase involved in phosphatidylinositol trimannoside biosynthesis in Mycobacterium tuberculosis; Kremer L et al.; Phosphatidyl-myo-inositol mannosides (PIMs), lipomannan (LM) and lipoarabinomannan (LAM) are an important class of bacterial factors termed modulins that are found in tuberculosis and leprosy . Although their structures are well established, little is known with respect to the molecular aspects of the biosynthetic machinery involved in the synthesis of these glycolipids . On the basis of sequence similarity to other glycosyltransferases and our previous studies defining an alpha-mannosyltransferase from Mycobacterium tuberculosis, named PimB {Schaeffer, Khoo, Besra, Chatterjee, Brennan, Belisle and Inamine (1999) J . Biol . Chem . 274, 31625-31631}, which catalysed the formation of triacyl (Ac(3))-PIM(2) (i.e . the dimannoside), we have identified a related gene from M . tuberculosis CDC1551, now designated pimC . The use of a cell-free assay containing GDP-{(14)C}mannose, amphomycin and membranes from Myobacterium smegmatis overexpressing PimC led to the synthesis of a new alkali-labile PIM product . Fast-atom-bombardment MS established the identity of the new enzymically synthesized product as Ac(3)PIM(3) (i.e . the trimannoside) . The results indicate that pimC encodes an alpha-mannosyltransferase involved in Ac(3)PIM(3) biosynthesis . However, inactivation of pimC in Myobacterium bovis Bacille Calmette-Guerin (BCG) did not affect the production of higher PIMs, LM and LAM when compared with wild-type M . bovis BCG, suggesting the existence of redundant gene(s) or an alternate pathway that may compensate for this PimC deficiency . Further analyses, which compared the distribution of pimC in a panel of M . tuberculosis strains, revealed that pimC was present in only 22% of the clinical isolates examined. Cell Mol Life Sci, 2002 Mar, 59(3), 410 - 6 Roles of Bacillus endospores in the environment; Nicholson WL; The occurrence and diverse roles of Bacillus spp . and their endospores in the environment is reviewed, with particular emphasis on soil ecology, host-symbiont and host-parasite interactions, and human exploitation of spores as biological control agents and probiotics. Appl Biochem Biotechnol, 2001 Spring, 91-93, 643 - 54 Characterization of cyclodextrin glycosyltransferase from Bacillus firmus strain no . 37; Matioli G et al.; The enzyme cyclodextrin glycosyltransferase (CGTase), EC 2.4.1.19, which produces cyclodextrins (CDs) from starch, was obtained from Bacillusfirmus strain no . 37 isolated from Brazilian soil and characterized in the soluble form using as substrate 100 g/L of maltodextrin in 0.05 M Tris-HCl buffer, 5 mM CaCl2, and appropriate buffers . Enzymatic activity and its activation energy were determined as a function of temperature and pH . The activation energy for the production of beta- and gamma-CD was 7.5 and 9.9 kcal/mol, respectively . The energy of deactivation was 39 kcal/mol . The enzyme showed little thermal deactivation in the temperature range of 35-60 degrees C, and Arrhenius-type equations were obtained for calculating the activity, deactivation, and half-life as a function of temperature . The molecular weight of the enzyme was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, giving 77.6 kDa . Results for CGTase activity as a function of temperature gave maximal activity for the production of beta-CD at 65 degrees C, pH 6.0, and 71.5 mmol of beta-CD/(min x mg of protein), whereas for gamma-CD it was 9.1 mmol of gamma-CD/(min mg of protein) at 70 degrees C and pH 8.0 . For long contact times, the best use of the enzymatic activity occurs at 60 degrees C or at a lower temperature, and the reaction pH may be selected to increase the yield of a desired CD. Prikl Biokhim Mikrobiol, 2002 Mar-Apr, 38(2), 161 - 5 {Protein and peptide factors from Bacillus sp.739 inhibit the growth of phytopathogenic fungi}; Shirokov AV et al.; Thermolabile peptides inhibiting the growth of Helminthosporium sativum, a facultative phytopathogen, have been isolated from the low-molecular-weight fraction of extracellular metabolites of the strain Bacillus sp . 739 . Paper chromatography of the fraction, followed by bioautography, revealed the presence of three components exhibiting antifungal activity . These components were separated by gel chromatography on Toyopearl HW-40 . SDS-PAGE (the Laemmli procedure) demonstrated that only one component was a protein (MW, approximately 14 kDa) . The other two substances were polypeptides with molecular weights less than 6 kDa each . The protein factor inhibited the growth of H . sativum with a minimum effective concentration of 0.1 to 0.2 mg/ml. Prikl Biokhim Mikrobiol, 2002 Mar-Apr, 38(2), 140 - 4 {Entomopathogenic bacteria Bacillus thuringiensis as producers of restriction endonucleases}; Puchkova LI et al.; A total of 115 collection strains of Bacillus thuringiensis, belonging to various subspecies, have been studied for the presence of DNA restriction-modification systems . Restriction endonucleases of 13 strains have been isolated and characterized . No considerable correlations between the taxonomic positions of the bacteria and the specificities of the endonucleases isolated have been detected . It is concluded that the enzymes with identical specificities are present in both the crystalliferous and acry