J Bacteriol, 2003 Jan, 185(2), 461 - 5 Bioenergetic properties of the thermoalkaliphilic Bacillus sp . strain TA2.A1; Olsson K et al.; The thermoalkaliphilic Bacillus sp . strain TA2.A1 was able to grow in pH-controlled batch culture containing a nonfermentable growth substrate from pH 7.5 to 10.0 with no significant change in its specific growth rate, demonstrating that this bacterium is a facultative alkaliphile . Growth at pH 10.0 was sensitive to the protonophore carbonyl cyanide m-chlorophenylhydrazone, suggesting that a proton motive force (Deltap) generated via aerobic respiration was an obligate requirement for growth of strain TA2.A1 . Strain TA2.A1 exhibited intracellular pH homeostasis as the external pH increased from 7.5 to 10.0; however, the maximum DeltapH generated over this pH range was only 1.1 units at an external pH of 9.5 . The membrane potential (Deltapsi) was maintained between -114 mV and -150 mV, and little significant change was observed over the pH range for growth . In contrast, the Deltap declined from -164 mV at pH 7.5 to approximately -78 mV at pH 10.0 . An inwardly directed sodium motive force (DeltapNa(+)) of -100 mV at pH 10.0 indicated that cellular processes (i.e., solute transport) dependent on a sodium gradient would not be affected by the adverse Deltap . The phosphorylation potential of strain TA2.A1 was maintained between -300 mV and -418 mV, and the calculated H(+)/ATP stoichiometry of the ATP synthase increased from 2.0 at pH 7.5 to 5.7 at pH 10.0 . Based on these data, vigorous growth of strain TA2.A1 correlated well with the DeltapNa(+), phosphorylation potential, and the ATP/ADP ratio, but not with Deltap . This communication represents the first report on the bioenergetics of an extremely thermoalkaliphilic aerobic bacterium. Int J Syst Evol Microbiol, 2002 Nov, 52(Pt 6), 2127 - 33 Two novel psychrotolerant species, Bacillus psychrotolerans sp . nov . and Bacillus psychrodurans sp . nov., which contain ornithine in their cell walls; Abd El-Rahman HA et al.; Eleven psychrotolerant Bacillus strains with ornithine as diamino acid in position 3 of the peptide side chain of the cell wall and a G+C range of 35.7-38.4 mol% were characterized taxonomically . DNA-DNA hybridization studies confirmed previously physiologically established groups . High DNA-binding values (> 70%) were found within groups I A (consisting of the type strain of Bacillus insolitus DSM 5(T) and Bacillus insolitus DSM 2272), I B (consisting of isolates 3H1(T0, 71H1, 84E1, 87H2 and 4H2) and I C (consisting of isolates 68E39T), 61E1, 4E3 and 67E1) . Low DNA-binding values (< 60%) were revealed between the three groups . Consequently, strains of groups I B and I C were considered as being representatives of new psychrotolerant species . For group I B strains the name Bacillus psychrotolerans sp . nov . is proposed with the type strain 3H1(T) (= DSM 11706(T) = NCIMB 13838(T)) and for group I C strains the name Bacillus psychrodurans sp . nov . is proposed with the type strain 68E3(T) (= DSM 11713(T) = NCIMB 13837(T)). Int J Syst Evol Microbiol, 2002 Nov, 52(Pt 6), 1985 - 9 Bacillus luciferensis sp . nov., from volcanic soil on Candlemas Island, South Sandwich archipelago; Logan NA et al.; Aerobic, endospore-forming bacteria were found in soil taken from an active fumarole on Lucifer Hill, Candlemas Island, South Sandwich archipelago . Amplified rDNA restriction analysis, SDS-PAGE, repetitive element primed-PCR (rep-PCR) and routine phenotypic tests suggested that six of the isolates represent a novel taxon, and 16S rDNA sequence comparisons support the proposal of a novel species, Bacillus luciferensis sp . nov., the type strain of which is strain LMG 18422(T) (= CIP 107105(T)). J Infect Dis, 2003 Jan 1, 187(1), 117 - 23 Epub 2002 Dec 02. Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guérin attenuation; Lewis KN et al.; The tuberculosis (TB) vaccine bacille Calmette-Guerin (BCG) is a live attenuated organism, but the mutation responsible for its attenuation has never been defined . Recent genetic studies identified a single DNA region of difference, RD1, which is absent in all BCG strains and present in all Mycobacterium tuberculosis (MTB) strains . The 9 open-reading frames predicted within this 9.5-kb region are of unknown function, although they include the TB-specific immunodominant antigens ESAT-6 and CFP-10 . In this study, RD1 was deleted from MTB strain H37Rv, and virulence of H37Rv:DeltaRD1 was assessed after infections of the human macrophage-like cell line THP-1, human peripheral blood monocyte-derived macrophages, and C57BL/6 mice . In each of these systems, the H37Rv:DeltaRD1 strain was strikingly less virulent than MTB and was very similar to BCG controls . Therefore, it was concluded that genes within or controlled by RD1 are essential for MTB virulence and that loss of RD1 was important in BCG attenuation. J Mol Biol, 2003 Jan 24, 325(4), 677 - 95 Lysyl-tRNA synthetase from Bacillus stearothermophilus: the Trp314 residue is shielded in a non-polar environment and is responsible for the fluorescence changes observed in the amino acid activation reaction; Takita T et al.; Three Trp variants of lysyl-tRNA synthetase from Bacillus stearothermophilus, in which either one or both of the two Trp residues within the enzyme (Trp314 and Trp332) were substituted by a Phe residue, were produced by site-directed mutagenesis without appreciable loss of catalytic activity . The following two phenomena were observed with W332F and with the wild-type enzyme, but not with W314F: (1) the addition of L-lysine alone decreased the protein fluorescence of the enzyme, but the addition of ATP alone did not; (2) the subsequent addition of ATP after the addition of excess L-lysine restored the fluorescence to its original level . Fluorometry under various conditions and UV-absorption spectroscopy revealed that Trp314, which was about 20A away from the lysine binding site and was shielded in a non-polar environment, was solely responsible for the fluorescence changes of the enzyme in the L-lysine activation reaction . Furthermore, the microenvironmental conditions around the residue were made more polar upon the binding of L-lysine, though its contact with the solvent was still restricted . It was suggested that Trp314 was located in a less polar environment than was Trp332, after comparison of the wavelengths at the peaks of fluorescence emission and of the relative fluorescence quantum yields . Trp332 was thought, based on the fluorescence quenching by some perturbants and the chemical modification with N-bromosuccinimide, to be on the surface of the enzyme, whereas Trp314 was buried inside . The UV absorption difference spectra induced by the L-lysine binding indicated that the state of Trp314, including its electrostatic environment, changed during the process, but Trp332 did not change . The increased fluorescence from Trp314 at acidic pH compared with that at neutral pH suggests that carboxylate(s) are in close proximity to the Trp314 residue. Int J Food Microbiol, 2003 Jan 26, 82(1), 71 - 9 Influence of pH and temperature on growth of Bacillus cereus in vegetable substrates; Valero M et al.; Bacillus cereus is a food-borne pathogen which most often contaminates foods of plant origin . Spores of psychrotrophic strains have the ability to germinate and grow at refrigeration temperatures in different vegetable substrates, such as carrot broth, zucchini broth, and cooked carrot puree . In some circumstances, factors such as pH, heat treatment, and storage temperature play a fundamental role in controlling the growth of these psychrotrophic strains and in extending the shelf life of refrigerated, minimally processed vegetable-based products in relation to pathogenic spore-forming bacteria . The combination of mild acidification (pH 5.0) and refrigeration (</=8 degrees C) inhibited B . cereus growth for at least 60 days in vegetable substrates similar to those mentioned above . This protection was maintained even when the temperature to which the food was exposed reached 12 degrees C . Psychrotrophic strains of B . cereus were inhibited in carrot broth by heating at 90 degrees C for 7.5 min, if the broth was refrigerated at a temperature of 8 degrees C or lower . If the vegetable product was exposed to temperatures of mild abuse (12 degrees C), it was necessary to implement a more drastic heat treatment (90 degrees C for 30 min). Indian J Pediatr, 2002 Nov, 69(11), 1003 - 5 Cutaneous bacillary angiomatosis; Asharaf M et al.; Bacillary angiomatosis is characterized by unique vascular lesions caused by infection with a small Gram staining bacillus of the genus Bartonella . It usually occurs in immunocompromised persons but can also occur in immunocompetent persons . We report a case of cutaneous bacillary angiomatosis in a 5-year-old immunocompetent child . He had infected lesions on the lips, after an injury, which was followed by lesions over the knees, buttocks, near the ankles and the elbows . Diagnosis was proved on histology . The lesions cleared after administration of erythromycin for 3 months . It is well to be aware of this condition in the context of increasing prevalence of AIDS. Expert Rev Anticancer Ther, 2002 Dec, 2(6), 667 - 70 Prognostic value of p53 overexpression in bladder tumors treated with Bacillus Calmette-Guerin; Peyromaure M et al.; Bacillus Calmette-Guerin intravesical therapy is the standard treatment of superficial bladder tumors at high risk of recurrence and progression to muscle-invasion disease . To date, there is no well established predictive factor of response to Bacillus Calmette-Guerin intravesical therapy . The prognostic value of p53 overexpression in bladder tumors is controversial . Most investigators have found no correlation between p53 status assessed before Bacillus Calmette-Guerin intravesical therapy and patient outcome . On the other hand, it is acknowledged that the persistence of p53 overexpression after Bacillus Calmette-Guerin intravesical therapy is predictive of progression in patients treated for carcinoma in situ . Since conflicting data have been reported, further evaluation of the impact of p53 overexpression in patients treated with Bacillus Calmette-Guerin intravesical therapy for bladder carcinoma is required. J Agric Food Chem, 2003 Jan 1, 51(1), 100 - 5 Enterotoxigenicity and cytotoxicity of Bacillus thuringiensis strains and development of a process for Cry1Ac production; Yang CY et al.; Bacillus thuringiensis is indistinguishable from Bacillus cereus except for the production of insecticidal crystal proteins (ICPs) . B . thuringiensis strains may show enterotoxin profiles and toxin levels similar to those of B . cereus strains isolated from food-poisoning cases . It is important for the food industry and farmers to consider that with the application of B . thuringiensis strains to crops, their spores may be introduced into the human food chain . In this study, 59 B . thuringiensis strains were assayed for their hemolysin BL (HBL) using a BCET-RPLA kit and their cytotoxicity to Chinese hamster ovary (CHO) cells . The enterotoxin titer was as high as that of B . cereus diarrheal-type strain ATCC 49064 . In an attempt to obtain a food safety strain for bioinsecticide use, in this study, a 3.5-kb cry1Ac DNA fragment was amplified with PCR from the total DNA of B . thuringiensis subsp . kurstaki CCRC 11502 and cloned into the Bacillus expression vector pHY300PLK . The alpha-amylase promoter, amyE, was then introduced into the promoter region and, afterward, the recombinant plasmid pHYe1Ac35 was introduced into a non-enterotoxigenic and non-cytotoxic B . thuringiensis subsp . kurstaki Tt14 strain . The transformant, without any detectable enterotoxigenicity or cytotoxicity, produced Cry1Ac toxin properly, and its insecticidal activity against Trichoplusia ni larvae was found to be satisfactory. J Gen Appl Microbiol, 2002 Oct, 48(5), 243 - 50 Application of the hypervariable region of the 16S rDNA sequence as an index for the rapid identification of species in the genus Alicyclobacillus; Goto K et al.; A comparison of the 16S rRNA gene (rDNA) sequences of seven type strains belonging to different Alicyclobacillus species (i.e., five validated species, one proposed species and one genomic species) suggested that the 5' end hypervariable region (259-273 bases in length) of 16S rDNA was specific for the respective type strains . Further phylogenetic analysis based on DNA sequences of the hypervariable region using 24 Alicyclobacillus strains revealed that the strains could be categorized into five species and the A . acidocaldarius-Alicyclobacillus genomic species 1 group . The hypervariable region was highly conserved among the five species: A . acidiphilus, A . acidoterrestris, A . cycloheptanicus, A . herbarius, and A . hesperidum . The strains in the A . acidocaldarius-Alicyclobacillus genomic species 1 group were subdivided into two clusters (Clusters I and II) based on DNA sequences in the hypervariable region . On the basis of phenotypic characteristics, chemotaxonomic and phylogenetic analyses, and DNA-DNA hybridization data, strains in Cluster I were grouped as Alicyclobacillus genomic species 1 and strains in Cluster II were re-identified as A . acidocaldarius, thereby demonstrating that the hypervariable regions were also highly conserved within these two species . These results suggest that as is the case with Bacillus, the hypervariable region is significantly species-specific in the genus Alicyclobacillus to distinguish Alicyclobacillus species by DNA sequence comparison of the hypervariable region. J Gen Appl Microbiol, 1999 Aug, 45(4), 149 - 153 Production of 2-phenylethylamine by decarboxylation of L-phenylalanine in alkaliphilic Bacillus cohnii; Hamana K et al.; Cellular polyamine fraction of alkaliphilic Bacillus species was analyzed by HPLC . 2-Phenylethylamine was found selectively and ubiquitously in the five strains belonging to Bacillus cohnii within 27 alkaliphilic Bacillus strains . A large amount of this aromatic amine was produced by the decarboxylation of L-phenylalanine in the bacteria and secreted into the culture medium . The production of 2-phenylethylamine may serve for the chemotaxonomy of alkaliphilic Bacillus. J Gen Appl Microbiol, 1999 Dec, 45(6), 283 - 287 In vitro degradation of keratin by two species of Bacillus; Lal S et al.; The ability of two species of Bacillus to degrade child's scalp hair, cow horn, cow hooves, and human nails in vitro under static conditions was studied by the determination of soluble sulphhydryl compounds as cysteine, disulphides as cystine, and release of extracellular keratinase along with changes in alkalinity of the culture filtrate . Child's scalp hair was found to be the most favored keratin substrate for Bacillus spp. J Gen Appl Microbiol, 1997 Dec, 43(6), 341 - 347 Cloning of a chitinase gene into Bacillus thuringiensis subsp . aizawai for enhanced insecticidal activity; Tantimavanich S et al.; Chitinase from a high producing strain (TP-1) of Bacillus licheniformis was used with B . thuringiensis subsp . aizawai (B.t.a.) in a combined larvicidal assay against the pest, Spodoptera exigua . With 10 mU of this chitinase, the LD(50) of B.t.a . was reduced by 7.6, 13.8 and 15 times on days 3, 5 and 7, respectively when compared to use of B.t.a . alone . In addition, a combination of chitinase (10 mU) and B.t.a . at a sub-lethal dose retarded growth and development of S . exigua . In preparation for transformation of B.t.a., the TP-1 chitinase gene was cloned in E . coli DH5alpha and sequenced to reveal a single open reading frame of 1,815 bp . This open reading frame encoded for a protein of 604 amino acids and a characteristic signal peptide sequence of 35 amino acids . The gene was subsequently introduced into B.t.a . where it was expressed constitutively . The transformed strain showed slightly improved activity against S . exigua when compared to the non-transformed strain . This was probably due to the low chitinase activity (15 mU/ml) of the transformant, which might be improved by further gene manipulation to overexpress enzyme production. J Gen Appl Microbiol, 1998 Apr, 44(2), 133 - 138 Seasonal distribution and characterization of Bacillus thuringiensis isolated from sericultural environments in Korea; Kim HS et al.; To isolate naturally occurring novel Bacillus thuringiensis strains, we investigated the distribution and characteristics of B . thuringiensis from samples of sericultural farms in various regions of Korea in the spring and fall . Fifty-four B . thuringiensis strains out of 164 samples and 34 B . thuringiensis strains out of 135 samples were isolated in the spring and fall, respectively . Seventy percent of the isolates in the spring and 15% in the fall were toxic to lepidopteran larvae . Dipteran-active isolates were rare (7% in spring and 3% in fall isolation) . Particularly, B . thuringiensis isolates, which are toxic to both Lepidoptera and Diptera, were widely distributed (19% in spring and 62% in fall isolation) . Non-toxic isolates were also found (4% in spring and 20% in fall isolation) . B . thuringiensis isolates in the sericultural farms represented 11 H serotypes; they were principally B . thuringiensis subsp . aizawai in the spring and kurstaki in the fall . B . thuringiensis isolates of serotypes 1, 3a, 3a3b, 4a4c, 6, 7 and 12 were toxic to Lepidoptera . Seventy isolates produced typical rhomboidal inclusions, and the remainder produced parasporal inclusions with various morphologies . PCR analysis using cryI gene type-specific primers showed that cryIAa and cryIC genes are frequently found in the spring and cryIAa gene is a predominant type in the fall . Toxicity, H serotype and the cryI gene contents of B . thuringiensis isolated from sericultural farms showed that distribution varied depending on the season. Neurology, 2002 Dec 24, 59(12), 1837 - 43 Immunization and MS: a summary of published evidence and recommendations; Rutschmann OT et al.; OBJECTIVE: To review the risk of MS exacerbations after infectious episodes potentially preventable by vaccination, and the risks and benefits of immunizing patients with MS . METHODS: The authors searched MEDLINE (1966 to January 2001; U.S . National Library of Medicine, Bethesda, MD), HealthSTAR, and Cumulative Index to Nursing and Allied Health Literature (CINAHL) database (Cinahl Information Systems, Glendale, CA) for English-language articles . Each study was summarized and rated for quality of evidence . Then feasible data were pooled and analyzed in meta-analysis . RESULTS: The risk of contracting common infectious diseases in patients with MS is not well established . There is strong evidence for an increased risk of MS exacerbations during weeks around an infectious episode . There is strong evidence against an increased risk of MS exacerbation after influenza immunization . There is no evidence that hepatitis B, varicella, tetanus, or Bacille Calmette-Guerin vaccines increase the risk of MS exacerbations . Insufficient evidence was found for other vaccines . CONCLUSIONS: Evidence supports 1) strategies to minimize the risk of acquiring infectious diseases that may trigger exacerbations of MS; and 2) the safety of using influenza, hepatitis B, varicella, tetanus, and Bacille Calmette-Guerin (BCG) vaccines in patients with MS. Antimicrob Agents Chemother, 2003 Jan, 47(1), 383 - 6 gyrA mutations in ciprofloxacin-resistant Bartonella bacilliformis strains obtained in vitro; Minnick MF et al.; We isolated and characterized mutants of Bartonella bacilliformis that are resistant to the fluoroquinolone antibiotic ciprofloxacin, which targets the A subunit of DNA gyrase . Mutants had single point mutations in the gyrA gene that changed either Asp-90 to Gly or Asp-95 to Asn and had 3- or 16-fold higher resistance, respectively, to ciprofloxacin than did wild-type B . bacilliformis . Asp-95 is homologous to Asp-87 of Escherichia coli GyrA and is a common residue mutated in fluoroquinolone-resistant strains of other bacteria . This is the first report of a mutation at an Asp-90 homologue, which corresponds to Asp-82 in E . coli GyrA. Clin Immunol, 2002 Dec, 105(3), 326 - 31 Oral recombinant Mycobacterium bovis bacillus Calmette-Guérin expressing HIV-1 antigens as a freeze-dried vaccine induces long-term, HIV-specific mucosal and systemic immunity; Kawahara M et al.; Induction of HIV-1-specific immune responses was evaluated using a recombinant BCG (rBCG) vector-based vaccine expressing HIV-1 Env V3 peptide (rBCG-pSOV3J1) . rBCG-pSOV3J1 was manufactured as a freeze-dried preparation based on good laboratory practice guidelines . Guinea pigs were immunized with the freeze-dried rBCG vaccine by oral administration to test the effectiveness of what is generally considered the most convenient and practical route for vaccination . While delayed-type hypersensitivity (DTH) skin reactions to purified protein derivative were not detected in any of the animals receiving oral rBCG-pSOV3J1, HIV-1 V3J1 antigen-specific DTH responses were detected in all of the immunized guinea pigs 1.5 years after immunization . In addition, significant proliferative responses against HIV-1 V3J1 antigen were measured in peripheral blood mononuclear cells and splenocytes from all animals receiving oral rBCG . Interestingly, intestinal intraepithelial lymphocytes from the animals also exhibited high levels of proliferative activity against HIV-1 V3J1 antigen . These results suggest that oral vaccination of guinea pigs with freeze-dried rBCG-pSOV3J1 induces high levels of functional T cells specific for HIV-1 antigens in both mucosal and systemic compartments and suggest that this approach has potential for use as a vaccine against HIV-1. Clin Immunol, 2002 Dec, 105(3), 296 - 303 Dendritic cells are decreased in blood and accumulated in granuloma in tuberculosis; Uehira K et al.; Immunity against tuberculosis consists of innate and adaptive immune responses . In this study, we investigated the dynamics of dendritic cells (DC), which are known to elicit a variety of immune responses, in patients with tuberculosis . CD11c(+) peripheral blood DC were decreased in patients with tuberculosis . Immunohistochemical analyses demonstrated that a number of fascin(+), CD11c(+), HLA-DR(+) DC were infiltrating the lymphocyte areas of the tuberculous granulomas (tubercles) . Immunohistochemical analyses also demonstrated that interferon-gamma-producing Th1 cells were increased in the tubercles of the patients, indicating the presence of Th1 polarization at least in the context of inflammatory tissues . In vitro coculture of autologous naive T cells with CD11c(+) or CD11c(-) DC pretreated with Bacillus Calmette Guerin augmented the production of Th1 cells . These findings suggested that the trafficking of DC from the peripheral blood into the tubercles causes a dominant Th1 balance and thus plays an essential role in the immunity against tuberculosis. J Immunol, 2003 Jan 1, 170(1), 463 - 9 Characterization of lung gamma delta T cells following intranasal infection with Mycobacterium bovis bacillus Calmette-Guérin; Dieli F et al.; The lungs are considered to have an impaired capacity to contain infection by pathogenic mycobacteria, even in the presence of effective systemic immunity . In an attempt to understand the underlying cellular mechanisms, we characterized the gammadelta T cell population following intranasal infection with Mycobacterium bovis bacillus Calmette-Guerin (BCG) . The peak of gammadelta T cell expansion at 7 days postinfection preceded the 30 day peak of alphabeta T cell expansion and bacterial count . The expanded population of gammadelta T cells in the lungs of BCG-infected mice represents an expansion of the resident Vgamma2 T cell subset as well as an influx of Vgamma1 and of four different Vdelta gene-bearing T cell subsets . The gammadelta T cells in the lungs of BCG-infected mice secreted IFN-gamma following in vitro stimulation with ionomycin and PMA and were cytotoxic against BCG-infected peritoneal macrophages as well as against the uninfected J774 macrophage cell line . The cytotoxicity was selectively blocked by anti-gammadelta TCR mAb and strontium ions, suggesting a granule-exocytosis killing pathway . Depletion of gammadelta T cells by injection of specific mAb had no effect on the subsequent developing CD4 T cell response in the lungs of BCG-infected mice, but significantly reduced cytotoxic activity and IFN-gamma production by lung CD8 T cells . Thus, gammadelta T cells in the lungs might help to control mycobacterial infection in the period between innate and classical adaptive immunity and may also play an important regulatory role in the subsequent onset of alphabeta T lymphocytes. Infect Immun, 2003 Jan, 71(1), 101 - 8 Oral delivery of Mycobacterium bovis BCG in a lipid formulation induces resistance to pulmonary tuberculosis in mice; Aldwell FE et al.; A lipid-based formulation has been developed for oral delivery of Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccine . The formulated M . bovis BCG was fed to BALB/c mice to test for immune responses and protection against M . bovis infection . The immune responses included antigen-specific cytokine responses, spleen cell proliferation, and lymphocyte-mediated macrophage inhibition of M . bovis . Oral delivery of formulated M . bovis BCG to mice induced strong splenic gamma interferon levels and macrophage inhibition of virulent M . bovis compared with results with nonformulated M . bovis BCG . Formulated oral M . bovis BCG significantly reduced the bacterial burden in the spleen and lungs of mice following aerosol challenge with virulent M . bovis . Our data suggest that oral delivery of formulated M . bovis BCG is an effective means of inducing protective immune responses against tuberculosis . Lipid-based, orally delivered mycobacterial vaccines may be a safe and practical method of controlling tuberculosis in humans and animals. Biophys J, 2002 Dec, 83(6), 3416 - 24 Bidirectional control of sphingomyelinase activity and surface topography in lipid monolayers; Fanani ML et al.; Lipid lateral organization is increasingly found to modulate membrane-bound enzymes . We followed in real time the reaction course of sphingomyelin (SM) degradation by Bacillus cereus sphingomyelinase (SMase) of lipid monolayers by epifluorescence microscopy . There is evidence that formation of ceramide (Cer), a lipid second messenger, drives structural reorganization of membrane lipids . Our results provide visual evidence that SMase activity initially alters surface topography by inducing phase separation into condensed (Cer-enriched) and expanded (SM-enriched) domains . The Cer-enriched phase grows steadily as the reaction proceeds at a constant rate . The surface topography derived from the SMase-driven reaction was compared with, and found to differ from, that of premixed SM/Cer monolayers of the same lipid composition, indicating that substantial information content is stored depending on the manner in which the surface was generated . The long-range topographic changes feed back on the kinetics of Smase, and the onset of condensed-phase percolation is temporally correlated with a rapid drop of reaction rate . These observations reveal a bidirectional influence and communication between effects taking place at the local molecular level and the supramolecular organization . The results suggest a novel biocatalytic-topographic mechanism in which a surface enzymatic activity can influence the function of amphitropic proteins important for cell function. J Food Prot, 2002 Dec, 65(12), 1930 - 6 Altered ability of Bacillus cereus spores to grow under unfavorable conditions (presence of nisin, low temperature, acidic pH, presence of NaCl) following heat treatment during sporulation; Faille C et al.; The effect of thermal treatment on the heat resistance of Bacillus cereus spores and their ability to germinate and grow under more or less adverse conditions during sporulation was investigated . Spores produced by sporulating cells subjected to a mild heat treatment (at a temperature 15 degrees C higher than the growth temperature) were more resistant to heat than were spores produced by untreated cells . Spore germination and growth (the lag time, the maximal growth rate, and the occurrence of a decrease in population) may be greatly affected by adverse environmental conditions brought about by the addition of nisin, low temperatures, acidic pHs, and, to a lesser extent, the addition of NaCl . Furthermore, heat treatments applied to sporulating cells or to mature spores induced a modification of the lag time (interaction of both treatments) . Therefore, mild heat treatments applied during sporulation may affect the heat resistance of spores and the ability of these spores to germinate under adverse conditions and may thus increase the risk associated with the presence of spores in lightly processed foods. J Antimicrob Chemother, 2003 Jan, 51(1), 107 - 12 Emergence of penicillin resistance among Fusobacterium nucleatum populations of commensal oral flora during early childhood; Nyfors S et al.; Penicillin resistance due to beta-lactamase production is surprisingly common among oral bacteria in childhood . Fusobacterium nucleatum, a Gram-negative anaerobic bacillus, is a member of the developing oral commensal flora . As part of the investigation on the emergence of oral bacterial resistance, the aim of the present study was to examine longitudinally the penicillin resistance among salivary F . nucleatum populations as related to age, day care attendance and sibling history, and exposure to antimicrobial agents . Altogether 1492 F . nucleatum isolates from saliva of 44 healthy infants followed at a study clinic at 2, 6, 12, 18 and 24 months of age were tested for beta-lactamase production . Furthermore, the 276 beta-lactamase-positive isolates were examined for their in vitro susceptibility to penicillin G by the NCCLS-approved agar dilution method . Statistical analysis of the associations between penicillin-resistant isolates and infants' age, day care attendance, number of siblings and their ear infections, and exposure to antimicrobial agents was performed by SPSS Windows Version 10 . The prevalence of infants harbouring beta-lactamase-producing F . nucleatum strains increased from 2% to 49% during the follow-up time . In nearly all cases beta-lactamase-producing F . nucleatum isolates were found simultaneously with beta-lactamase-negative isolates . Most beta-lactamase-producing isolates (80%) showed an MIC of > or =8 mg/L . In conclusion, the prevalence of infants harbouring penicillin-resistant F . nucleatum due to beta-lactamase production increased with age and usage of antimicrobial agents during the first year of life. J Appl Microbiol, 2003, 94(1), 60 - 4 Properties of Bacillus thuringiensis isolated from bank voles; Swiecicka I et al.; AIMS: To assess the properties of B . thuringiensis naturally occurring in the intestines of bank voles . METHODS AND RESULTS: Seventeen Bacillus thuringiensis strains, exhibiting typical growth on selective medium for the B . cereus group and characterized by the ability to produce parasporal crystals, were isolated from bank voles trapped in the Lomza Landscape Park of the Narew River Valley (north-east Poland) . All isolates were characterized by pulsed field gel electrophoresis (PFGE) of chromosomal DNA and SDS polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins . Six pulsotypes were found with PFGE typing, using SmaI or NotI as restriction enzymes . Significant differences in chromosome size, ranging from 2.4 to 4.2 Mb for the B . thuringiensis strains studied, were noted . Strain heterogeneity in pulsotypes was also reflected by the similarity of whole-cell protein profiles of the strains . Environmental isolates and reference strains grouped at 71% similarity according to SDS-PAGE data and at 84% on the basis of biochemical tests . CONCLUSIONS: B . thuringiensis from intestines of bank voles demonstrated an important level of heterogeneity . The comparison of PFGE profiles and SDS-PAGE of whole-cell protein patterns may be useful to evaluate the relationship between B . thuringiensis isolates . SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented in this paper may help to explain the diversity of B . thuringiensis. Zhonghua Nan Ke Xue, 2002, 8(4), 266 - 9 {The establishment of testicular fibrosis model in Wistar rats}; Wang T et al.; OBJECTIVES: To establish the testicular fibrosis model in rats . METHODS: Wistar rats were divided into control group(n = 12) and model group(n = 40) randomly . Testicular fibrosis model was built with the classical method of establishing experimental autoimmune orchitis (EAO) combined with injecting Bacille Calmette-Guerin (BCG) into left testis . RESULTS: The incidence rate of EAO and the rate of testicular fibrosis were 100%, 11.1% and 100%, 81.5% at 80, 140 days after the first infection, respectively . CONCLUSIONS: The model of rat testicular fibrosis was established successfully. Clin Infect Dis, 2003 Jan 1, 36(1), 16 - 23 Epub 2002 Dec 13. Improved detection of Mycobacterium tuberculosis in Peruvian children by use of a heminested IS6110 polymerase chain reaction assay; Montenegro SH et al.; A novel heminested IS6110 polymerase chain reaction (PCR) assay was evaluated as a tool for diagnosing tuberculosis in 222 children . In an analysis of 392 specimens (gastric aspirates, nasopharyngeal aspirates, and sputum samples), results of PCR were compared with those of 3 culture methods, acid-fast bacillus (AFB) staining, and clinical assessment by the Stegen-Toledo score . The sensitivity of PCR (67%) was comparable to that of the 3-culture method (71%) and was significantly higher than that of Lowenstein-Jensen culture (54%) or AFB stain (42%) for children with highly probable tuberculosis . PCR detection rates for culture-positive specimens were 100% for smear-positive samples and 76.7% for smear-negative samples . The specificity of PCR was 100% in control children . Compared with culture, PCR demonstrated a sensitivity of 90.4%, a positive predictive value of 89%, a specificity of 94%, and a negative predictive value of 95% (kappa=.85) . With clinical assessment as the standard, PCR had a sensitivity of 71%, a positive predictive value of 92%, a specificity of 95%, and a negative predictive value of 79% (kappa=.67) . PCR is a rapid and sensitive method for the early diagnosis of pediatric tuberculosis. Arch Virol, 2002 Dec, 147(12), 2393 - 404 Sequence analysis of an Australian isolate of sugarcane bacilliform badnavirus; Geijskes RJ et al.; The genome of an Australian isolate of Sugarcane bacilliform virus (SCBV-IM) was cloned, sequenced and analysed . The genome consisted of 7687 nucleotides and contained three open reading frames which were similar in size and organisation to those of other badnaviruses . SCBV-IM was found to be most similar to the SCBV-Morocco isolate with amino acid sequence similarity of 91.4 %, 83.8 % and 85.3 % in the ORF I, II and III coding regions, respectively . Phylogenetic analysis of the SCBV-IM ORF III deduced amino acid sequence showed that SCBV isolates were more closely related to each other than to other badnaviruses . Amplification of SCBV sequences from three different sugarcane varieties revealed considerable variability in the viral populations, both within single infected plants as well as between infected plants, suggesting that the SCBV isolates sequenced to date may not be representative of the range of virus variability. Gene, 2002 Nov 13, 301(1-2), 13 - 20 Characterization of two regulatory genes of the mercury resistance determinants from TnMERI1 by luciferase-based examination; Huang CC et al.; The broad-spectrum mercury resistance transposon, TnMERI1, of Bacillus megaterium strain MB1, contains three proposed operator/promoter (O/P) transcriptional start sites and two regulatory genes (merR1 and merR2) . A series of luciferase (lux)-based transcriptional fusion plasmids were studied in Escherichia coli to show that both merR1 and merR2 gene products repressed transcription from O/PmerB3, O/PmerR1, and O/PmerR2 under uninduced conditions . Derepression occurred when the merR1 gene was present and Hg(2+) functioned as an inducer . In the presence of organomercurial compounds, basal transcription of merB3 was needed to produce inorganic Hg(2+) as the inducer of expression regulated by MerR1 at O/PmerB3 . The presence of merR2 repressed transcription from all three O/Pmer sites under both non-induced conditions and when inorganic Hg(2+) or organomercurials were added . These results show that MerR1 functions as a repressor in the absence of Hg(2+) and as an activator in the presence of Hg(2+), while MerR2 functions as a repressor. Zhonghua Jie He He Hu Xi Za Zhi, 2002 Oct, 25(10), 595 - 7 {A study on the use of mycobacterium vaccine in the treatment of tuberculous pleurisy}; Zhou X et al.; OBJECTIVE: To study the effects of immunotherapy with mycobacterium vaccine in patients with tuberculous pleurisy . METHODS: The time for effusion resolution and lymphocyte counts in the effusions were observed in patients with tuberculous pleurisy who received anti-tuberculous therapy and thoracentesis (group A) or anti-tuberculous therapy and thoracentesis plus intramuscularly administered polysaccharide nucleic acid fraction of Bacillus Calmette Guerin (PSN-BCG,) (group S) . In situ end-labeling technique of fragment DNA was used to detect the apoptosis of lymphocytes in the effusions . RESULTS: The average resolution times in group A and group S were 21.2 days and 28.7 days respectively (P < 0.01) . The lymphocyte counts in the two groups after 1 week, 2 week, and 3 week therapy were 1.6 x 10(9)/L (A) and 2.1 x 10(9)/L (S) (P < 0.01), 0.83 x 10(9)/L and 1.52 x 10(9)/L (P < 0.01), 0.55 x 10(9)/L and 1.16 x 10(9)/L (P < 0.01) respectively . The average apoptotic half-time was 94 h (A) and 124 h (S) (P < 0.01), 84 h and 123 h (P < 0.01), 79 h and 120 h (P < 0.01) respectively . CONCLUSIONS: Mycobacterium vaccine was found to prolong lymphocyte activation, inhibit lymphocyte apoptosis, and delay the resolution of pleural effusions . It was found not helpful as an adjunct therapy for tuberculous pleurisy. Clin Lab Med, 2002 Dec, 22(4), 937 - 62 Laboratory diagnosis of Bartonella infections; Agan BK et al.; Bartonella species are pathogens of emerging and reemerging significance, causing a wide array of clinical syndromes . In North America and Europe, they are increasingly recognized as a cause of culture negative endocarditis, neuroretinitis, and disease among homeless, HIV-infected, and other immunosuppressed individuals . In South America, bartonellosis continues to plague those in endemic regions and poses a significant threat to travelers in these areas . As the clinician is increasingly faced with these illnesses, which may be difficult to diagnose, laboratory techniques to confirm or refute the diagnosis are becoming increasingly important . Culture methods have improved over the past decade demonstrating increased sensitivity, but still require prolonged periods before isolation of the organism . Specimen handling, media selection, and growth conditions all may affect results and must be optimized in order to provide the highest likelihood of recovering the organism . Pure culture of the bacteria not only provides morphologic information, but also provides material for further diagnostic testing . Work with liquid media, which may provide a more rapid means of cultivation has shown some promise and should continue to be pursued . Improved blood culture techniques were a primary factor in the discovery of Bartonella endocarditis and continued improvements will likely demonstrate further clinical insights . Serologic testing for B henselae infections has become the cornerstone of clinical diagnosis, replacing the skin test that was poorly standardized and posed a potential risk to the patient . Immunofluorescence assays have been well characterized and validated in clinical trials, however they are not universally available . Vero cell cocultivated antigens appear to provide higher sensitivity and specificity when compared with agar-derived antigens . IFA assays are inherently difficult to perform, requiring significant expertise to provide reproducible results . On the contrary, enzyme immunoassays offer ease of use and a high level of reproducibility, however ideal antigens for use in the diagnosis of Bartonella infections have not been clearly identified . Continued work to define antigenic targets of the human response to infection and incorporation of these into a widely available EIA will provide a cost-effective tool for the clinician and epidemiologist alike . Due to the close phylogenetic relationship of B henselae and B quintana, differentiation between these species by serologic means may prove difficult . Molecular techniques including PCR offer high sensitivity and specificity, rapid availability of information, and the ability to differentiate Bartonella organisms at the highest level . Results of studies to date are promising and as methods are refined it will be important to conduct clinical studies to define the role of these assays . In disseminated Bartonella infections such as bacillary angiomatosis, peliosis, endocarditis, and urban trench fever, PCR currently offers the ability to establish the diagnosis when other tests may be unrevealing . For CSD, this technique should be used as a confirmatory technique when the diagnosis is unclear by other means . PCR analysis of blood specimens offers a minimally invasive approach to diagnosis, but clinical data are scarce and further studies are needed . As DNA microarrays move into the clinical arena, specific hybridization probes may allow improved identification and differentiation of Bartonellae at the molecular level. J Med Microbiol, 2003 Jan, 52(Pt 1), 69 - 74 Production by Bacillus pumilus (MSH) of an antifungal compound that is active against Mucoraceae and Aspergillus species: preliminary report; Bottone EJ et al.; A compound produced by Bacillus pumilus (MSH) that inhibits Mucoraceae and Aspergillus species is described . Fungicidal activity was demonstrated by lawn-spotting and by diffusion through 0.45 microm Millipore membranes placed on 5 % sheep-blood agar, nutrient agar, trypticase soy agar and Mueller-Hinton agar, followed by spore inoculation of the bacterium-free underlying agar surface . With either technique, zones of fungal inhibition correlated with the zone of haemolysis produced by B . pumilus (MSH) . The active compound inhibited Mucor and Aspergillus spore germination and aborted elongating hyphae, presumably by inducing a cell-wall lesion . Antifungal activity was stable in agar for a minimum of 8 days, resistant to Pronase degradation, and partially inactivated by chloroform exposure and at pH 5.6 . Its molecular mass was determined by diffusion through dialysis membrane to be 500-3000 Da . Attempts at further isolation of the compound have proven unsuccessful to date. Biochim Biophys Acta, 2002 Dec 23, 1567(1-2), 113 - 22 Estimation of the radius of the pores formed by the Bacillus thuringiensis Cry1C delta-endotoxin in planar lipid bilayers; Peyronnet O et al.; Pore formation constitutes a key step in the mode of action of Bacillus thuringiensis delta-endotoxins and various activated Cry toxins have been shown to form ionic channels in receptor-free planar lipid bilayers at high concentrations . Multiple conductance levels have been observed with several toxins, suggesting that the channels result from the multimeric assembly of a variable number of toxin molecules . To test this possibility, the size of the channels formed by Cry1C was estimated with the non-electrolyte exclusion technique and polyethylene glycols of various molecular weights . In symmetrical 300 mM KCl solutions, Cry1C induced channel activity with 15 distinct conductance levels ranging from 21 to 246 pS and distributed in two main conductance populations . Both the smallest and largest conductance levels and the mean conductance values of both populations were systematically reduced in the presence of polyethylene glycols with hydrated radii of up to 1.05 nm, indicating that these solutes can penetrate the pores formed by the toxin . Larger polyethylene glycols had little effect on the conductance levels, indicating that they were excluded from the pores . Our results indicate that Cry1C forms clusters composed of a variable number of channels having a similar pore radius of between 1.0 and 1.3 nm and gating synchronously. Intern Med, 2002 Nov, 41(11), 990 - 2 Pulmonary sequestration associated by Mycobacterium intracellulare infection; Shiota Y et al.; A case of a 29-year-old woman with intralobar pulmonary sequestration infected with Mycobacterium intracellulare is presented . A chest CT scan revealed a density in the posterior segment of the left lower lobe, and an acid-fast bacillus sputum culture yielded Mycobacterium intracellulare . After 3 months of treatment with clarithromycin, streptomycin, rifampicin and ethambutol, the patient underwent partial resection of the left lower lobe . At the 6-month follow-up the patient's clinical status is excellent . A review of the literature revealed only three case reports of pulmonary sequestration associated with Mycobacterium avium-intracellulare complex infection. J Mol Evol, 2002 Dec, 55(6), 632 - 7 Simultaneous horizontal gene transfer of a gene coding for ribosomal protein l27 and operational genes in Arthrobacter sp; Garcia-Vallve S et al.; Phylogenetic analysis of bacterial L27 ribosomal proteins showed that, against taxonomy, the L27 protein from the Actinobacteria Arthrobacter sp . clusters with protein sequences from the Bacillus group . The L27 gene clusters in the Arthrobacter sp . genome with six genes responsible for creatinine and sarcosine degradation . Phylogenetic analyses of orthologue proteins encoded by three of these genes also showed a phylogenetic relationship with Bacillus species . Comparisons between the synonymous codon usage of the Arthrobacter sp . genes and those from complete genomes showed that Arthrobacter genes encoding the L27 ribosomal protein and the proteins responsible for the degradation of creatinine and sarcosine have a codon usage that is more similar to that of Bacillus species than that of Arthrobacter . We suggest that the Arthrobacter sp . genes encoding the L27 ribosomal protein and the proteins responsible for the degradation of creatinine and sarcosine were acquired simultaneously through horizontal gene transfer from an unknown Bacillus species. Int J Lepr Other Mycobact Dis, 2002 Sep, 70(3), 174 - 81 Combined 12-month WHO/MDT MB regimen and Mycobacterium w . vaccine in multibacillary leprosy: a follow-up of 136 patients; Kaur I et al.; A total of 136 patients with BI > or = 2 having been followed up for at least 2 years or more were included in the analyses . Seventy-seven out of 136 patients had completed three years follow up . All patients were given WHO/MDT MB regimen for 12 months and additionally 4 doses of Mycobacterium w . vaccine at 3-month intervals . The age of the patients varied from 6 to 77 years (mean 34 +/- 11.3 years) and they had the disease varying from 3 months to 7 years (mean = 1.9 +/- 1.4 years) . The mean of the BI before starting treatment was 3.6 +/- 1.3 . At the end of 2 years follow-up, a total of 54 patients out of the 136 (39.7%) had become smear-negative . A larger proportion of patients, 39/46 (84.8%) with BI of < or = 3 had become smear-negative, whereas, only 10/32 (31.3%) patients with BI between 3.1 to 4 and 5/58 (8.6%) highly bacillated patients having initial BI > 4 had become smear-negative at the end of 2 years . Out of the 77 patients who were available for follow up at 3 years, 30/33 (90.9%) patients with BI of < or = 3, 15/20 (75%) patients with BI between 3.1 to 4 and 13/24 (54.2%) patients having initial BI > 4, respectively, had attained smear negativity . Reactions occurred more frequently after 6 months of therapy and over a period of time their frequency gradually decreased, however, they continued to occur even two years after RFT . During the course of MDT and thereafter in follow up 4.6% and 1.3% of the patients developed new deformities or an increase in the existing grade of deformities, respectively . Three relapses (2 in LL and 1 in BL) occurred in patients having initial BI of > 4 . One patient relapsed in the second year and the other two relapsed in the third year of follow up and were successfully treated with reintroduction of the same MDT MB regimen . Local ulceration healing with scar formation and regional lymphadenopathy were the only local reactions to the vaccine seen in 47/136 (34.5%) patients . All the patients showed histopathological improvement in the form of a gradual reduction of granuloma fraction . Although the results of this limited period follow up are satisfactory, a long-term follow-up in larger number of patients will settle the issue of safety and efficacy of shortened MDT MB regimen and the place of immunotherapy with M . w . vaccine in multibacillary patients. J Gen Appl Microbiol, 2001 Oct, 47(5), 263 - 267 Production and characterization of tannase from Bacillus cereus KBR9; Mondal KC et al.; A tannase-producing soil bacteria has been isolated and identified as Bacillus cereus . It can degrade tannic acid and produce maximum tannase (0.22 U/ml) at stationary phases of growth (24 h) . Maximum growth and enzyme production occurred with initial medium pH of 4.5-5.0 . Partial purified tannase showed optimum activity at pH 4.5 and 40 degrees C . It remains stable up to 30 degrees C and pH 4.5 to 5.0 . The enzyme is salt tolerant, stable up to 2 m of NaCl and retains 82% original activity in 3 m. J Gen Appl Microbiol, 2000 Feb, 46(1), 1 - 8 Application of the partial 16S rDNA sequence as an index for rapid identification of species in the genus Bacillus; Goto K et al.; A comparison of 16S rRNA gene (rDNA) sequences was made among type strains of 69 Bacillus species approved in the International Journal of Systematic Bacteriology (IJSB) until 1998 . The results suggested that 5' end region (approx . 275 bp) was the hypervariant region (HV region) in the gene and was highly specific for each type strain . Furthermore, a sequence analysis of the HV region of Bacillus strains revealed that this region was highly conserved within the species . These results indicate that the HV region is a useful index for the identification or grouping of Bacillus species. J Eur Acad Dermatol Venereol, 2002 Nov, 16(6), 612 - 4 Bacillary angiomatosis of the scalp in a human immunodeficiency virus-negative patient; Kayaselcuk F et al.; Bacillary angiomatosis (BA) is a recently recognized bacterial infectious disease that is mainly seen in patients with the acquired immunodeficiency syndrome . BA has rarely been reported in immunocompetent individuals . A case of BA of the scalp in an immunocompetent patient, who was human immunodeficiency virus seronegative, is reported . The importance of differential diagnosis of this lesion is emphasized. Parazitologiia, 2002 Sep-Oct, 36(5), 337 - 44 {Cytopathological effect of Bacillus thuringiensis israelensis endotoxins on the intestines of Aedes aegypti mosquito larvae}; Zalunin IA et al.; The dynamics of pathological changes in the intestine of Aedes aegypti larvae under the influence of toxins Cry11A and Cry4B produced by Bacillus thuringiensis israelensis was studied by means of electron microscope . Most significant ultrastructure changes in the intestine of the second instar larvae were observed in the midgut . The cytoplasm of cells disintegrated, and elongated lacunae appeared . The number of microvilli decreased, or they disappeared in the result of destruction . The peritrophic membrane displaced to the lumen of midgut . Any changes in epithelial cells and cuticle in time of foregut and hindgut were not observed in a comparison to control . The toxin Cry4B caused the most effective destruction of the midgut epithelium. Biochem Biophys Res Commun, 2003 Jan 3, 300(1), 102 - 6 Differential expression of CYP102 in Bacillus megaterium by 17-beta-estradiol and 4-sec-butylphenol; Rowley CW et al.; Previously we have reported the induction of CYP102 in Bacillus megaterium by 17beta-estradiol (E2) and 4-sec-butylphenol (4-sBP) . Electrophoretic mobility shift assay analyses demonstrated that E2 and 4-sBP both cause a dose-dependent disassociation of the Bm3R1 repressor protein from its binding site on the operator sequence of the CYP102 gene . Equimolar combinations of E2 and 4-sBP demonstrated additive induction of CYP102 compared to equivalent samples of E2 and 4-sBP added alone . Two gene constructs were used in this investigation . One construct designated BMC143 contained the entire regulatory region of CYP102 . The other gene construct, designated BMA45, had the "Barbie box" sequence deleted . While the induction of CYP102 by 4-sBP was much higher in the BMC 143 construct, E2 induced CYP102 in both constructs to the same extent . This difference in induction of CYP102 by these two inducers indicates that they act at different sites, either on the Bm3R1 repressor protein or on positive regulatory sites, or that they act, in part, through different mechanisms. FEMS Microbiol Lett, 2002 Dec 17, 217(2), 263 - 7 Analysis of Bacillus megaterium lipolytic system and cloning of LipA, a novel subfamily I.4 bacterial lipase; Ruiz C et al.; The lipolytic system of Bacillus megaterium 370 was investigated, showing the existence of at least two secreted lipases and a cell-bound esterase . A gene coding for an extracellular lipase was isolated and cloned in Escherichia coli . The cloned enzyme displayed high activity on short to medium chain length (C(4)-C(8)) substrates, and poor activity on C(18) substrates . On the basis of amino acid sequence homology, the cloned lipase was classified into subfamily I.4 of bacterial lipases . Water Sci Technol, 2002, 46(10), 247 - 54 Simultaneous production of biopesticide and alkaline proteases by Bacillus thuringiensis using sewage sludge as a raw material; Tyagi RD et al.; The simultaneous production of Bacillus thuringiensis (Bt) based biopesticide and proteases was studied using synthetic medium and wastewater sludge as a raw material . The studies were conducted in shake flask and computer controlled 15-L capacity fermentors . Measuring viable cell and spore counts, entomotoxicity and protease activity monitored the progress of the biopesticide production process . A higher viable cell count and spore count was observed in synthetic Soya medium, however, higher entomotoxicity and protease activity were observed in wastewater sludge medium . Thus, the wastewater sludge is a better raw material than commercial Soya medium for the biopesticides and enzyme production . The maximum entomotoxicity and protease activity observed in the fermentor was 9,332 IU/microL and 4.58 IU/mL, respectively . The proteases produced by Bt were also characterised . Two types of proteases were detected; neutral proteases with pH optimum 7.0 and alkaline proteases with pH optimum 10-11 . Further, two types of alkaline proteases were detected; one having a pH and temperature optimum at 10 and 50 degrees C while the other at 11 and 70 degrees C . The protease thermal stability was found to increase in the presence of CaCl2, indicating the proteases were metalloproteases. Chem Commun (Camb), 2002 Dec 7, (23), 2860 - 1 Biosynthesis of aminoglycoside antibiotics: cloning, expression and characterisation of an aminotransferase involved in the pathway to 2-deoxystreptamine; Huang F et al.; The gene btrR from Bacillus circulans has been cloned and expressed and shown to produce a protein which catalyses the transamination of 2-deoxy-scyllo-inosose to give 2-deoxy-scyllo-inosamine, an intermediate in the biosynthesis of 2-deoxystreptamine. J Urol, 2003 Jan, 169(1), 357 - 60 A phase I study of intravesical suramin for the treatment of superficial transitional cell carcinoma of the bladder; Uchio EM et al.; PURPOSE: Suramin is a polysulfonated naphthylurea that inhibits proliferation and DNA synthesis of transitional cell carcinoma cell lines . Its large molecular size and negative charge inhibit bladder absorption, making suramin an excellent candidate for intravesical chemotherapy . Intravesical suramin was evaluated in a phase I study to define dose limiting toxicity and systemic absorption, determine a starting dose and regimen for phase II studies and provide a preliminary assessment of in vivo antitumor activity . MATERIALS AND METHODS: Intravesical suramin treatment was administered in 9 patients with histologically identified transitional cell carcinoma (Tcis, Ta or T1) in whom at least 1 course of standard intravesical chemotherapy (bacillus Calmette-Guerin, thiotepa or mitomycin C) had failed . Suramin was administered once weekly for 6 weeks . Patients were treated in groups of 3 using a 60 cc volume and intrapatient dose escalation schedule . Suramin doses of 0.3 to 614.4 mg./ml . were administered intravesically . The last group was treated with the same weekly dose for 6 weeks . RESULTS: The 9 patients underwent 54 treatments with suramin . Plasma suramin concentration after treatment was 1.9 to 38.0 microg./ml . and was not related to treatment dose . The dose escalation phase was limited by the solubility of suramin in solution . Complications included self-limited bladder spasms (less than 24 hours) in 4 of 54 treatments (7%) and new or worsening vesicoureteral reflux in 3 ureters (17%) . Another patient who was treated after the Foley balloon was inflated in the urethra experienced bladder spasms, skin flushing and fever (39C) . Mean bladder capacity before and after treatment was 600 and 540 ml., respectively . At followup 7 patients had stage Ta tumors and 2 had carcinoma in situ . CONCLUSIONS: An intravesical suramin dose of 153 mg./ml was defined as a safe treatment parameter with acceptable plasma concentrations and minimal side effects . Phase II studies are needed to assess the antitumor activity of suramin in patients with transitional cell carcinoma of the bladder. J Urol, 2003 Jan, 169(1), 96 - 100; discussion 100 A retrospective analysis of 153 patients treated with or without intravesical bacillus Calmette-Guerin for primary stage T1 grade 3 bladder cancer: recurrence, progression and survival; Shahin O et al.; PURPOSE: We retrospectively evaluated the long-term outcome in patients with newly diagnosed stage T1 grade 3 bladder cancer treated with transurethral resection with or without intravesical bacillus Calmette-Guerin (BCG) . MATERIALS AND METHODS: Of 153 patients with a median age of 67 years (range 36 to 88) and a male-to-female ratio of 4:1 we treated 92 with transurethral bladder resection and additional BCG, and 61 with transurethral bladder resection alone . BCG was administered intravesically as 120 mg . BCG Pasteur F dissolved in 50 ml . saline, retained for up to 2 hours weekly for 6 weeks and repeated as necessary . RESULTS: Median followup was 5.3 years (range 0.4 to 18.2) . Disease recurred in 70% of the patients treated with BCG and in 75% treated with transurethral resection alone . Median time to recurrence was 38 and 22 months for BCG and resection alone (p = 0.19) . Tumor progressed in 33% of patients with BCG and in 36% with resection alone . Deferred cystectomy was performed in 29% of the patients with BCG and in 31% with resection alone . Overall and disease specific survival did not differ significantly . CONCLUSIONS: Our results suggest that intravesical BCG therapy after transurethral bladder resection for stage T1 grade 3 bladder cancer may delay the time to recurrence and cystectomy but it does not substantially alter the final outcome . Our findings reflect the rule of 30% for stage T1 grade 3 cancer, namely approximately 30% of patients never have recurrence, 30% ultimately die of metastatic disease and 30% require deferred cystectomy. J Urol, 2003 Jan, 169(1), 90 - 5 Intravesical bacillus Calmette-Guerin versus mitomycin C for superficial bladder cancer: a formal meta-analysis of comparative studies on recurrence and toxicity; Bohle A et al.; PURPOSE: We compare the therapeutic efficacy and toxicity of intravesical bacillus Calmette-Guerin (BCG) with mitomycin C on recurrence of stages Ta and T1 bladder carcinoma . MATERIALS AND METHODS: Combined published and unpublished data from comparative studies on BCG versus mitomycin C for superficial bladder carcinoma considering possible confounding factors were analyzed . Odds ratio (OR) and its 95% CI were used as primary effect size estimate . Toxicity data were evaluated descriptively . RESULTS: In 11 eligible clinical trials 1,421 patients were treated with BCG and 1,328 were treated with mitomycin C . Within the overall median followup time of 26 months 38.6% of the patients in the BCG group and 46.4% of those in the mitomycin C group had tumor recurrence . In 7 of 11 studies BCG was significantly superior to mitomycin C, in 3 studies no significant difference was found, while in 1 study mitomycin C was significantly superior to BCG . An overall statistically significant superiority of BCG versus mitomycin C efficacy in reducing tumor recurrence was detected (OR 0.56, 95% CI 0.38 to 0.84, p = 0.005) . In the subgroup treated with BCG maintenance all 6 individual studies showed a significant superiority of BCG over mitomycin C (OR 0.43, 95% CI 0.35 to 0.53, p <0.001) . In 4 of the 5 studies with reported data on toxicity BCG associated cystitis was significantly more frequent than in the mitomycin C group (53.8% versus 39.2%) . The combined cystitis OR was 1.81 (95% CI 1.48 to 2.23, p <0.001) . The OR for cystitis in the BCG maintenance group did not significantly differ from that in the nonmaintenance therapy group . CONCLUSIONS: The results suggest superiority of BCG over mitomycin C for prevention of tumor recurrences in the combined data and particularly in the BCG maintenance treatment subgroup, irrespective of the actual (intermediate or high) tumor risk status . The toxicity with BCG is higher but does not differ between BCG maintenance and nonmaintenance groups. Eur Urol, 2002 Dec, 42(6), 542 - 6 Beware the BCG failures: a review of one institution's results; Lockyer CR et al.; OBJECTIVE: To review the outcome of all superficial transitional cell (TCC) bladder cancer treated with intravesical Bacille Calmette-Guerin (BCG) at one institution and, in particular, the prognosis for those patients who gained little benefit from BCG therapy . PATIENTS AND METHODS: The notes of 122 patients treated with BCG over a nine-year period were reviewed . The following details were recorded: time of diagnosis; time of decision to treat with BCG; results of cystoscopies before and after BCG; duration of follow up; time of progression if occurred, mortality and cause of death . RESULTS: Complete follow up data was available for 112 patients . At a median follow up of 23 months (range 3-107) 57 patients (51%) remained free of tumour, 30 (27%) had progressed and 18 (16%) had died of transitional cell carcinoma . There was a significant association between a positive initial check cystoscopy and subsequent progression (p<0.001) and disease specific mortality (p<0.001) . Of the 35 patients who had a positive cystoscopy after BCG treatment 21 (60%) progressed and 14 (40%) died of transitional cell carcinoma compared with 9 (12%) and 4 (5%) of the 77 with a negative cystoscopy . Adjusted odds ratios for progression and death from TCC for patients with a positive initial check cystoscopy were 21 and 13, respectively . CONCLUSION: In our series the patients found to have tumour at the initial check cystoscopy following intravesical BCG had a poor prognosis . This should be remembered when considering treatment options and counselling patients . Follow up of all BCG patients need to be rigorous and protocols would help to unify the treatment patients receive. J Steroid Biochem Mol Biol, 2002 Oct, 82(2-3), 257 - 61 Influence of substituents at C11 on hydroxylation of progesterone analogs by Bacillus sp; Das S et al.; Transformation of progesterone analogs viz., progesterone, 11 alpha-, 11 beta-hydroxyprogesterones and 11-ketoprogesterone by Bacillus sp . is reported . Both progesterone and 11-ketoprogesterone were hydroxylated while the C(11) epimeric alcohols of progesterone remained unaltered under the conditions used . The major bioconverted products obtained from progesterone and 11-ketoprogesterone were characterized as 6 beta- and 14 alpha-hydroxyprogesterones and 14 alpha-hydroxy-11-ketoprogesterone respectively by mass and NMR spectra . The conversion of 11-ketoprogesterone to its 14 alpha-hydroxy derivative by microbe is unprecedented and novel . Moreover, hydroxylation at 6 beta- and 14 alpha-positions of progesterone by Bacillus sp . is significant . In conclusion, the present data showed that the substituents at 11-position of steroid play important role on hydroxylation by microbe. J Biol Chem, 2003 Feb 28, 278(9), 7663 - 73 Epub 2002 Dec 09. Crystal structure of Bacillus sp . GL1 xanthan lyase, which acts on the side chains of xanthan; Hashimoto W et al.; Xanthan lyase, a member of polysaccharide lyase family 8, is a key enzyme for complete depolymerization of a bacterial heteropolysaccharide, xanthan, in Bacillus sp . GL1 . The enzyme acts exolytically on the side chains of the polysaccharide . The x-ray crystallographic structure of xanthan lyase was determined by the multiple isomorphous replacement method . The crystal structures of xanthan lyase and its complex with the product (pyruvylated mannose) were refined at 2.3 and 2.4 A resolution with final R-factors of 17.5 and 16.9%, respectively . The refined structure of the product-free enzyme comprises 752 amino acid residues, 248 water molecules, and one calcium ion . The enzyme consists of N-terminal alpha-helical and C-terminal beta-sheet domains, which constitute incomplete alpha(5)/alpha(5)-barrel and anti-parallel beta-sheet structures, respectively . A deep cleft is located in the N-terminal alpha-helical domain facing the interface between the two domains . Although the overall structure of the enzyme is basically the same as that of the family 8 lyases for hyaluronate and chondroitin AC, significant differences were observed in the loop structure over the cleft . The crystal structure of the xanthan lyase complexed with pyruvylated mannose indicates that the sugar-binding site is located in the deep cleft, where aromatic and positively charged amino acid residues are involved in the binding . The Arg(313) and Tyr(315) residues in the loop from the N-terminal domain and the Arg(612) residue in the loop from the C-terminal domain directly bind to the pyruvate moiety of the product through the formation of hydrogen bonds, thus determining the substrate specificity of the enzyme. Urol Oncol, 2002 Mar-Apr, 7(2), 67 - 72 Differential C-erbB-2 and VEGF expression following BCG immunotherapy in superficial papillary transitional cell carcinoma of the bladder; Morgan BE et al.; Bacillus Calmette Guerin (BCG) is generally regarded as an effective immunotherapy for superficially invasive papillary transitional cell carcinoma of the bladder . The exact mechanism(s) which underlie its efficacy are unknown . As C-erbB-2 oncoprotein and vascular endothelial growth factor (VEGF) have been shown to be over-expressed in TCC of the bladder, it has been postulated that they may be important in its pathogenesis . The purpose of this study was to 1.) differentially evaluate the effect of BCG immunotherapy in treated and untreated cohorts on the immunohistochemical expression of C-erbB-2 and VEGF in formalin-fixed paraffin-embedded sections of superficial and superficially invasive (Stage Ta-T1) transitional cell carcinoma of the bladder . Immunolabeling intensity was assessed independently by two pathologists and reported as a mean labeling index . The results confirm previous studies that 1.) both c-erbB-2 and VEGF are over-expressed in these tumors MLI = 90.1 and 45.7 respectively, 2.) that VEGF is an early and sensitive indicator of TCC, and 3.) that BCG has a salutary effect on papillary TCC, 66% vs . 89% recurrence rate, P = .04 . Our findings show that 1.) C-erbB-2 expression is decreased in patients tumors which show response to BCG (45.7 to 38.5), P = 0.15, 2.) that BCG administration has no effect on the expression of VEGF . While the decrement in c-erbB-2 immunostaining observed in those patients who received BCG contrasts with the increase in c-erbB-2 immunolabeling observed in patients who did not receive BCG, the differences were not statistically significant and could reflect tumor grade or stage regression associated with BCG therapy . However, this study suggests that BCG differentially influences the expression of C-erbB-2 and VEGF. Trans R Soc Trop Med Hyg, 2002 Sep-Oct, 96(5), 549 - 50 High rate of Bartonella henselae infection in HIV-positive outpatients in Johannesburg, South Africa; Frean J et al.; The emerging opportunistic pathogen Bartonella henselae has a wide range of clinical presentation, which includes, particularly, bacillary angiomatosis . This non-random pilot survey of outpatients attending HIV clinics in Johannesburg, South Africa, sampled 188 patients, in whom there was a 10% prevalence of Bartonella bacteraemia, as determined by nested polymerase chain reaction. J Biol Chem, 2003 Feb 28, 278(9), 7310 - 9 Epub 2002 Dec 06. Tracking the putative biosynthetic precursors of oxygenated mycolates of Mycobacterium tuberculosis . Structural analysis of fatty acids of a mutant strain deviod of methoxy- and ketomycolates; Dinadayala P et al.; Disruption of the mma4 gene (renamed hma) of Mycobacterium tuberculosis has yielded a mutant strain defective in the synthesis of both keto- and methoxymycolates, with an altered cell-wall permeability to small molecules and a decreased virulence in the mouse model of infection (Dubnau, E., Chan, J., Raynaud, C., Mohan, V . P., Laneelle, M . A., Yu, K., Quemard, A., Smith, I., and Daffe, M . (2000) Mol . Microbiol . 36, 630-637) . Assuming that the mutant would accumulate the putative precursors of the oxygenated mycolates of M . tuberculosis, a detailed structural analysis of mycolates from the hma-inactivated strain was performed using a combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, proton NMR spectroscopy, and chemical degradation techniques . These consisted most exclusively of alpha-mycolates, composed of equal amounts of C(76)-C(82) dicyclopropanated (alpha(1)) and of C(77)-C(79) monoethylenic monocyclopropanated (alpha(2)) mycolates, the double bond being located at the "distal" position . In addition, small amounts of cis-epoxymycolates, structurally related to alpha(2)-mycolates, was produced by the mutant strain . Complementation of the hma-inactivated mutant with the wild-type gene resulted in the disappearance of the newly identified mycolates and the production of keto- and methoxymycolates of M . tuberculosis . Introduction of the hma gene in Mycobacterium smegmatis led to the lowering of diethylenic alpha mycolates of the recipient strain and the production of keto- and hydroxymycolates . These data indicate that long-chain ethylenic compounds may be the precursors of the oxygenated mycolates of M . tuberculosis . Because the lack of production of several methyltransferases involved in the biosynthesis of mycolates is known to decrease the virulence of the tubercle bacillus, the identification of the substrates of these enzymes should help in the design of inhibitors of the growth of M . tuberculosis. Mem Inst Oswaldo Cruz, 2002 Oct, 97(7), 1041 - 8 Study of the safety, immunogenicity and efficacy of attenuated and killed Leishmania (Leishmania) major vaccines in a rhesus monkey (Macaca mulatta) model of the human disease; Amaral VF et al.; We have compared the efficacy of two Leishmania (Leishmania) major vaccines, one genetically attenuated (DHFR-TS deficient organisms), the other inactivated {autoclaved promastigotes (ALM) with bacillus Calmete-Guerin (BCG)}, in protecting rhesus macaques (Macaca mulatta) against infection with virulent L . (L.) major . Positive antigen-specific recall proliferative response was observed in vaccinees (79% in attenuated parasite-vaccinated monkeys, versus 75% in ALM-plus-BCG-vaccinated animals), although none of these animals exhibited either augmented in vitro gamma interferon (IFN-gamma) production or positive delayed-type hypersensitivity (DTH) response to the leishmanin skin test prior to the challenge . Following challenge, there were significant differences in blastogenic responses (p < 0.05) between attenuated-vaccinated monkeys and naive controls . In both vaccinated groups very low levels of antibody were found before challenge, which increased after infective challenge . Protective immunity did not follow vaccination, in that monkeys exhibited skin lesion at the site of challenge in all the groups . The most striking result was the lack of pathogenicity of the attenuated parasite, which persisted in infected animals for up to three months, but were incapable of causing disease under the conditions employed . We concluded that both vaccine protocols used in this study are safe in primates, but require further improvement for vaccine application. Anal Biochem, 2002 Dec 15, 311(2), 103 - 18 Radical-generating coordination complexes as tools for rapid and effective fragmentation and fluorescent labeling of nucleic acids for microchip hybridization; Kelly JJ et al.; DNA microchip technology is a rapid, high-throughput method for nucleic acid hybridization reactions . This technology requires random fragmentation and fluorescent labeling of target nucleic acids prior to hybridization . Radical-generating coordination complexes, such as 1,10-phenanthroline-Cu(II) (OP-Cu) and Fe(II)-EDTA (Fe-EDTA), have been commonly used as sequence nonspecific "chemical nucleases" to introduce single-strand breaks in nucleic acids . Here we describe a new method based on these radical-generating complexes for random fragmentation and labeling of both single- and double-stranded forms of RNA and DNA . Nucleic acids labeled with the OP-Cu and the Fe-EDTA protocols revealed high hybridization specificity in hybridization with DNA microchips containing oligonucleotide probes selected for identification of 16S rRNA sequences of the Bacillus group microorganisms.We also demonstrated cDNA- and cRNA-labeling and fragmentation with this method . Both the OP-Cu and Fe-EDTA fragmentation and labeling procedures are quick and inexpensive compared to other commonly used methods . A column-based version of the described method does not require centrifugation and therefore is promising for the automation of sample preparations in DNA microchip technology as well as in other nucleic acid hybridization studies. Mycorrhiza, 2002 Dec, 12(6), 313 - 6 Epub 2002 Sep 24. Inoculation of field-established mulberry and papaya with arbuscular mycorrhizal fungi and a mycorrhiza helper bacterium; Mamatha G et al.; The effects of soil inoculation with arbuscular mycorrhizal (AM) fungi and a mycorrhiza helper bacterium (MHB) were investigated on mulberry and papaya plants already established in the field . Ten-year-old mulberry plants (var . M5) were inoculated with Glomus fasciculatum and 1.5-year-old papaya plants (var . Solo) were inoculated with a mixed culture of G . mosseae and G . caledonium with or without Bacillus coagulans at two levels of P fertilizer . Growth, P uptake, yield and AM colonization levels were monitored . Leaf yield in mulberry and fruit yield in papaya were minimal in uninoculated plants given 50% recommended P . However, crop yields of both mulberry and papaya inoculated with AM fungi alone or together with the bacterium and given 50% recommended P were statistically on a par with that of uninoculated plants given 100% recommended P . As inoculation of B . coagulans increased mycorrhiza levels in AM fungal-inoculated plants, this may be included in the class of MHB . Thus, mulberry and papaya already established in the field may respond to AM inoculation and MHB may increase symbiosis development by efficient AM fungi. Appl Microbiol Biotechnol, 2002 Dec, 60(4), 381 - 95 Epub 2002 Oct 12. An overview on fermentation, downstream processing and properties of microbial alkaline proteases; Gupta R et al.; Microbial alkaline proteases dominate the worldwide enzyme market, accounting for a two-thirds share of the detergent industry . Although protease production is an inherent property of all organisms, only those microbes that produce a substantial amount of extracellular protease have been exploited commercially . Of these, strains of Bacillus sp . dominate the industrial sector . To develop an efficient enzyme-based process for the industry, prior knowledge of various fermentation parameters, purification strategies and properties of the biocatalyst is of utmost importance . Besides these, the method of measurement of proteolytic potential, the selection of the substrate and the assay protocol depends upon the ultimate industrial application . A large array of assay protocols are available in the literature; however, with the predominance of molecular approaches for the generation of better biocatalysts, the search for newer substrates and assay protocols that can be conducted at micro/nano-scale are becoming important . Fermentation of proteases is regulated by varying the C/N ratio and can be scaled-up using fed-batch, continuous or chemostat approaches by prolonging the stationary phase of the culture . The conventional purification strategy employed, involving e.g., concentration, chromatographic steps, or aqueous two-phase systems, depends on the properties of the protease in question . Alkaline proteases useful for detergent applications are mostly active in the pH range 8-12 and at temperatures between 50 and 70 degrees C, with a few exceptions of extreme pH optima up to pH 13 and activity at temperatures up to 80-90 degrees C . Alkaline proteases mostly have their isoelectric points near to their pH optimum in the range of 8-11 . Several industrially important proteases have been subjected to crystallization to extensively study their molecular homology and three-dimensional structures. Proc Natl Acad Sci U S A, 2002 Dec 24, 99(26), 16581 - 6 Epub 2002 Dec 03. Bacillus thuringiensis-toxin resistance management: stable isotope assessment of alternate host use by Helicoverpazea; Gould F et al.; Data have been lacking on the proportion of Helicovera zea larvae that develop on noncotton host plants that can serve as a refuge from selection pressure for adaptation to transgenic cotton varieties that produce a toxin from the bacterium Bacillus thuringiensis . We found that individual H . zea moths that develop as larvae on cotton and other plants with C3 physiology have a different ratio of 13C to 12C than moths that develop on plants with C4 physiology, such as corn . We used this finding in determining the minimum percentage of moths that developed on noncotton hosts in two cotton-growing areas . Our results indicate that local corn can serve as a refuge for H . zea in midsummer . Our results contrast dramatically with the prevailing hypothesis that the large majority of late-season moths are produced from larvae feeding on cotton, soybean, and other C3 plants . Typically, <50% of moths captured in August through October have isotope ratios indicative of larval feeding on C3 plants . In one October sample, 100% of the moths originated from C4 hosts even though C4 crops were harvested at least 1 mo earlier, and no common wild C4 hosts were available . These findings support other research indicating that many late-season H . zea moths captured in Louisiana and Texas are migrants whose larvae developed on corn in more northern locations . Our isotope data on moths collected in Texas early in the season indicate that the majority of overwintering H . zea do not originate from cotton-feeding larvae and may be migrants from Mexico . Non-Bt corn in Mexico and the U.S . corn belt appears to serve as an important refuge for H . zea. Cell, 2002 Nov 27, 111(5), 747 - 56 Sensing small molecules by nascent RNA: a mechanism to control transcription in bacteria; Mironov AS et al.; Thiamin and riboflavin are precursors of essential coenzymes-thiamin pyrophosphate (TPP) and flavin mononucleotide (FMN)/flavin adenine dinucleotide (FAD), respectively . In Bacillus spp, genes responsible for thiamin and riboflavin biosynthesis are organized in tightly controllable operons . Here, we demonstrate that the feedback regulation of riboflavin and thiamin genes relies on a novel transcription attenuation mechanism . A unique feature of this mechanism is the formation of specific complexes between a conserved leader region of the cognate RNA and FMN or TPP . In each case, the complex allows the termination hairpin to form and interrupt transcription prematurely . Thus, sensing small molecules by nascent RNA controls transcription elongation of riboflavin and thiamin operons and possibly other bacterial operons as well. Lett Appl Microbiol, 2002, 35(6), 523 - 7 Optimization of a fermentation medium for the production of Penicillin G acylase from Bacillus sp; Rajendhran J et al.; AIMS: Optimization of Penicillin G acylase (PAC) production from a novel isolate of Bacillus sp . METHODS: Fermentation medium for PAC production was optimized using a two-level fractional factorial design with seven components . RESULTS: A maximum production of 9.5 U ml(-1) of PAC was obtained in an optimized medium containing (g l(-1): K2HPO4, 1.0; MgSO4.7H2O, 0.1; CaCl2.2H2O, 0.1; PAA, 2.0; tryptone, 5.0; yeast extract, 3.0; and sucrose, 50.0 . SIGNIFICANCE AND IMPACT OF THE STUDY: The two-step medium optimization resulted in a twofold increase in PAC production . Since the strain Bacillus sp . PGS10 produces a high level of PAC, it could be a potential candidate for industrial production of PAC. Immunology, 2002 Dec, 107(4), 500 - 6 Phagocytosis of bacille Calmette-Guérin-infected necrotic macrophages induces a maturation phenotype and evokes antigen-presentation functions in dendritic cells; Goldmann O et al.; The interaction of pathogens with dendritic cells (DCs) seems to play a critical role in the initiation of the immune response . Tissue damage and induction of an inflammatory reaction are events frequently associated with the progression of the infection . Although DCs are very efficient at phagocytosing pathogens, the capacity of these cells to uptake microbes from a necrotic environment has not yet been proven . Here we have investigated the ability of murine bone marrow-derived DCs to maturate and acquire antigen-presentation functions when cocultured with bacille Calmette-Guerin (BCG)-infected necrotic macrophages . Immature DCs exhibited a prominent capacity to ingest necrotic material as demonstrated by flow cytometry analysis and confocal microscopy . Furthermore, after exposure to BCG-infected necrotic macrophages, DCs underwent phenotypic changes, including the up-regulation of maturation specific markers (major histocompatibility complex class II, CD40, CD80, and CD86) and the capacity to stimulate antigen-specific CD4+ T cells with higher efficiency than when they were directly infected with a similar number of bacteria . Antigen presentation following phagocytosis of BCG-infected necrotic macrophages was demonstrated by their ability to stimulate in vitro proliferation and interferon-gamma production of antigen-specific CD4+ T cells . These results suggest that the functional changes occurring in DCs after interaction with a pathogen can be favoured when the encounter takes place in a necrotic environment and it may constitute an important mechanism for the amplification of class II-restricted immune responses induced during infection. Virus Res, 2002 Dec, 90(1-2), 269 - 74 Genomic instability of prawn white spot bacilliform virus (WSBV) and its association to virus virulence; Lan Y et al.; Prawn White Spot Bacilliform Virus (WSBV) is a major pathogen that causes prawn diseases . In this study, we examined the sequence of WSBV genome DNA in the shrimp Penaeus japonicus, P . vannamei, P . Monodon, P . chinensis and Metapenaeus ensis through successive PCR amplification of the DNA fragments in the whole WSBV genome . We found a sequence deletion hotspot in the WSBV genome that is 305107 bp in length . The sizes of the deleted fragments were 4.6, 4.8 or 8.1 kbp depending on the species of prawn . Since the mortality of shrimp infected by the intact WSBV was always significantly higher than that of shrimp infected by DNA fragment-deleted WSBV, we suggest that this deletion be somehow linked to the virulence of the virus itself . This result may lead to the discovery of the molecular mechanism of the pathogenicity of WSBV in shrimps. Res Microbiol, 2002 Nov, 153(9), 605 - 9 Regulation of crystal protein biosynthesis by Bacillus thuringiensis: II . Effects of carbon and nitrogen sources; Icgen Y et al.; The regulation of crystal protein production in Bacillus thuringiensis 81 by sources of carbon and nitrogen was investigated . The highest titers of toxin were obtained on sucrose, lactose and inulin which also supported sporulation . Whey and molasses were also potential carbon substrates for toxin production . Other carbohydrates including glucose, glycerol, maltose, starch and dextrin yielded lower amounts of toxin . Nitrogen sources were found to exert the most profound controls . Peptone was the best organic nitrogen source, supporting optimum production and sporulation as well as high cell density . The formation of CryI and CryII toxin proteins was found to be differentially regulated by the inorganic nitrogenous compounds incorporated into the medium. Res Microbiol, 2002 Nov, 153(9), 599 - 604 Regulation of crystal protein biosynthesis by Bacillus thuringiensis: I . Effects of mineral elements and pH; Icgen Y et al.; Crystal protein synthesis by a local isolate of Bacillus thuringiensis was monitored and compared in association with growth and sporulation in media differing in mineral element content . Mg and Cu were the most important metals for the biosynthesis of 135 kDa and 65 kDa toxin components in that the former was essential and the latter was greatly stimulatory at 10(-6) to 10(-7) M concentration . Also the inclusion of Mn favored toxin production at concentrations ranging from 3 x 10(-4) to 10(-5) M . The omission of Zn and Ca had no effect on toxin formation . Crystal protein synthesis and sporulation did not generally seem to be co-regulated by the minerals as these processes responded differently to mineral levels . There was no evidence for suppression of biosynthesis by inorganic phosphate over a range of 3 to 100 mM . Crystal protein production was more efficient in buffered medium, especially when the initial pH was adjusted to 6.5. Acta Crystallogr D Biol Crystallogr, 2002 Dec, 58(Pt 12), 2198 - 200 Epub 2002 Nov 23. The 2.2 A resolution structure of thermolysin (TLN) crystallized in the presence of potassium thiocyanate; Gaucher JF et al.; A new crystallization protocol for thermolysin (EC 3.4.24.27) from Bacillus thermoproteolyticus is presented . After dissolving the protein in the presence of KSCN, which avoids the use of DMSO and CsCl, crystals were obtained following the salting-in method . Crystal cell parameters are isomorphous with those previously reported from DMSO/CsCl mixtures . The new SCN(-) crystal structure has been analyzed . It shows the presence of one thiocyanate ion in the catalytic site and several rearrangements in the S(1) and S(2) subsites . These results are in agreement with the measurements of Inouye et al . {(1998), J . Biochem . (Tokyo), 123, 847-852}, who observed in solution that the solubility of TLN, which is particularly poor in low ionic strength solutions, increases dramatically in the presence of several neutral salts . The results reported here suggest possible explanations for the solubility increase and for the inhibitory effects of high SCN(-) concentrations on thermolysin activity. Acta Crystallogr D Biol Crystallogr, 2002 Dec, 58(Pt 12), 2175 - 6 Epub 2002 Nov 23. Crystallization and preliminary X-ray diffraction analysis of a thermostable D-hydantoinase from the mesophilic Bacillus sp . AR9; Agrawal V et al.; D-hydantoinase catalyzes the conversion of DL-hydantoin derivatives to the corresponding optically pure N-carbamoyl amino acids, the first step in the industrial preparation of optically pure amino acids using biological catalysts . A thermostable D-hydantoinase from the mesophilic bacteria Bacillus sp . AR9 has been crystallized in three different crystal forms . The hexagonal faced crystals were the best looking, but did not diffract . One of the crystal forms is star-shaped and appeared to be twinned, but diffracted as a single crystal to a resolution of 2.3 A . These crystals belong to space group P6(4) and have unit-cell parameters a = b = 129.55, c = 102.86 A, alpha = beta = 90, gamma = 120 degrees. J Clin Microbiol, 2002 Dec, 40(12), 4750 - 2 Performance of the microscopic observation drug susceptibility assay in drug susceptibility testing for Mycobacterium tuberculosis; Park WG et al.; The drug susceptibility testing performance of a broth-based method with microscopic reading of bacillary growth, the microscopic observation drug susceptibility (MODS) assay, was compared to that of the reference 7H10 agar method of proportion by using 53 isolates of Mycobacterium tuberculosis from persons at risk for multidrug-resistant TB . For isoniazid (0.1 micro g/ml) and rifampin (2.0 micro g/ml), there was 100% agreement between MODS results read at day 11 and the reference method . Levels of agreement for ethambutol tested at 2.5 and 7.5 micro g/ml were 70 and 58%, respectively . Levels of agreement for streptomycin tested at 2.0 and 6.0 micro g/ml were 77 and 51%, respectively . For isoniazid and rifampin drug susceptibility testing, MODS is as accurate as and more rapid than the reference method. Emerg Infect Dis, 2002 Nov, 8(11), 1327 - 33 Tuberculosis-related deaths within a well-functioning DOTS control program; Garcia-Garcia Mde L et al.; To describe the molecular epidemiology of tuberculosis (TB)-related deaths in a well-managed program in a low-HIV area, we analyzed data from a cohort of 454 pulmonary TB patients recruited between March 1995 and October 2000 in southern Mexico . Patients who were sputum acid-fast bacillus smear positive underwent clinical and mycobacteriologic evaluation (isolation, identification, drug-susceptibility testing, and IS6110-based genotyping and spoligotyping) and received treatment from the local directly observed treatment strategy (DOTS) program . After an average of 2.3 years of follow-up, death was higher for clustered cases (28.6 vs . 7%, p=0.01) . Cox analysis revealed that TB-related mortality hazard ratios included treatment default (8.9), multidrug resistance (5.7), recently transmitted TB (4.1), weight loss (3.9), and having less than 6 years of formal education (2) . In this community, TB is associated with high mortality rates. J Appl Microbiol, 2002, 93(6), 937 - 43 Sterilization of plastic containers using electron beam irradiation directed through the opening; Cleghorn DA et al.; AIMS: Studies were performed to demonstrate the efficacy of a novel electron beam irradiation system for the on-line sterilization of polymeric containers using energetic electrons directed through the container opening . METHODS AND RESULTS: The distribution of dosage delivered during conveyance beneath the electron beam treatment system was determined for two sizes (8 or 16 ounces) of high-density polyethylene (HDPE) blow-moulded bottles . The biological effects of treatment were then determined using Bacillus pumilus ATCC 27142 spores inoculated as 10 microl droplets dried (i) onto the surface of flat coupons of the high-density polyethylene bottle material, and (ii) onto the region of lowest delivered dose (the bottom side wall) within each bottle . The inactivation obtained was determined by examining the level of survival after swab recovery of the inoculated spores, with reference to the level recovered from untreated control samples . CONCLUSIONS: The inactivation of B . pumilus spores during treatment was reproducible and proportional to the applied dose . In each instance, the logarithm of the surviving fraction of spores was well fitted by linear regression models and in good general agreement with values reported for inactivation of the same organism using gamma irradiation . SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this work demonstrate the potential of a new physical method capable of high throughput for in-line sterilization of polymeric containers . The ability of the process to eliminate the washing step involved with traditional chemical sterilants, such as hydrogen peroxide, greatly reduces the size and environmental impact of these systems. J Appl Microbiol, 1997 Feb, 82(2), 253 - 8 Natural occurrence of Bacillus thuringiensis on cabbage foliage and in insects associated with cabbage crops; Damgaard PH et al.; Bacillus thuringiensis was isolated from the phylloplane of organically grown cabbage in one field during two growth seasons (1992-93) . The frequency of B . thuringiensis varied between 0.02 and 0.67 of the total B . cereus/B . thuringiensis population, with an average of 0.11 . Characterization of the B . thuringiensis isolates from foliage showed that the majority (64% of 150 isolates) belonged to serovar kurstaki, had bipyramidal crystals and toxicity towards Pieris brassicae and/or Trichoplusia ni . Other serovars were also found on the foliage but occurred at very low frequencies (one to three isolates of each serovar) . Bacillus thuringiensis was also isolated from insects associated with the cabbage crop (Pieris rapae (Lep.), Delia radicum (Dip.), Syrphidae ribesii (Dip.) and Aleochara bilineata (Col.)), which were collected alive at different developmental stages in the same field . Serologically these isolates were assigned to the serovars kurstaki, aizawai, tochigiensis, colmeri and indiana/colmeri. J Gen Physiol, 2002 Dec, 120(6), 845 - 53 The cation selectivity filter of the bacterial sodium channel, NaChBac; Yue L et al.; The Bacillus halodurans voltage-gated sodium-selective channel (NaChBac) (Ren, D., B . Navarro, H . Xu, L . Yue, Q . Shi, and D.E . Clapham . 2001b . SCIENCE: 294:2372-2375), is an ideal candidate for high resolution structural studies because it can be expressed in mammalian cells and its functional properties studied in detail . It has the added advantage of being a single six transmembrane (6TM) orthologue of a single repeat of mammalian voltage-gated Ca(2+) (Ca(V)) and Na(+) (Na(V)) channels . Here we report that six amino acids in the pore domain (LESWAS) participate in the selectivity filter . Replacing the amino acid residues adjacent to glutamatic acid (E) by a negatively charged aspartate (D; LEDWAS) converted the Na(+)-selective NaChBac to a Ca(2+)- and Na(+)-permeant channel . When additional aspartates were incorporated (LDDWAD), the mutant channel resulted in a highly expressing voltage-gated Ca(2+)-selective conductance. Appl Environ Microbiol, 2002 Dec, 68(12), 6410 - 5 Existence of a true phosphofructokinase in Bacillus sphaericus: cloning and sequencing of the pfk gene; Alice AF et al.; Some strains of Bacillus sphaericus are entomopathogenic to mosquito larvae, which transmit diseases, such as filariasis and malaria, affecting millions of people worldwide . This species is unable to use hexoses and pentoses as unique carbon sources, which was proposed to be due to the lack of glycolytic enzymes, such as 6-phosphofructokinase (PFK) . In this study, PFK activity was detected and the pfk gene was cloned and sequenced . Furthermore, this gene was shown to be present in strains belonging to all the homology groups of this heterogeneous species, in which PFK activity was also detected . A careful sequence analysis revealed the conservation of different catalytic and regulatory residues, as well as the enzyme's phylogenetic affiliation with the family of allosteric ATP-PFK enzymes. Vaccine, 2002 Dec 13, 21(3-4), 158 - 66 Combined intrarectal/intradermal inoculation of recombinant Mycobacterium bovis bacillus Calmette-Guérin (BCG) induces enhanced immune responses against the inserted HIV-1 V3 antigen; Kawahara M et al.; The development of a successful recombinant Mycobacterium bovis bacillus Calmette-Guerin (rBCG) vector-based vaccine for human immunodeficiency virus type 1 (HIV-1) requires the induction of high levels of HIV-1-specific immunity while at the same time maintaining immunity to tuberculosis . To examine a combined vaccination strategy for enhancement of immune responses specific for HIV-1, guinea pigs were inoculated with either a single or combination intradermal (i.d.), intrarectal (i.r.) and intranasal (i.n.) administration of rBCG-pSOV3J1 which secretes a chimeric protein of HIV-1 V3J1 peptide and alpha-antigen . Significant level of delayed-type hypersensitivity to both V3J1 peptide and tuberculin was induced in guinea pigs inoculated with human doses of rBCG-pSOV3J1 by a combination of intrarectal and intradermal routes . Guinea pigs inoculated by combined routes also had significantly higher titers of HIV-1-specific serum IgG and IgA compared with those animals immunized only intrarectally, which led to the enhanced neutralization activity against HIV-1(MN) . In addition, the induction of high levels of IFNgamma and interleukin-2 (IL-2) mRNA in PBMC, splenocytes, and intraepithelial lymphocytes from the immunized animals was detected until at least 110 weeks post-inoculation . These results suggest that enhanced immune responses specific for HIV-1 are efficiently induced by combined intrarectal and intradermal immunization with rBCG-HIV, and antigen-specific Th1-type memory cells are maintained for more than 2 years in the immunized animals . Thus, inoculation with rBCG-HIV by combined routes represents an effective vaccination strategy to elicit high levels of HIV-1-specific immune responses. Lepr Rev, 2002 Sep, 73(3), 254 - 61 Anti-leprosy protective vaccination of rhesus monkeys with BCG or BCG plus heat-killed Mycobacterium leprae: lepromin skin test results; Gormus BJ et al.; Groups of rhesus monkeys (RM) were vaccinated and boosted with living Mycobacterium bovis Bacillus Calmette-Guerin (BCG) or BCG + low dose (LD) heat-killed Mycobacterium leprae (HKML) or high dose (HD) HKML or were unvaccinated . Animals vaccinated with BCG + LD and HD HKML were lepromin skin tested 2 weeks after boosting . All groups were lepromin tested 37 and 46 months after challenge with live M . leprae . Fernandez (72 h) and Mitsuda (28 day) responses were recorded . Ten of 10 rhesus monkeys in each of the two BCG + HKML-vaccinated groups significantly converted to strong positive Fernandez status within 2 weeks of boosting, compared to one of six positives in the unvaccinated unchallenged normal control group . Both BCG + HKML groups were significantly protected from clinical leprosy . Six of 10 in each of the two BCG + HKML groups significantly converted to Mitsuda positivity within 2 weeks of boosting compared to zero of six in the normal control group . The sizes of the Mitsuda responses were larger in the LD group than the HD HKML vaccinated/boosted group, suggesting suppression by vaccination with higher doses of HKML in combination with BCG . Fernandez responses were negative in normal RM as well as in the unvaccinated, ML-challenged group and the BCG-vaccinated, ML-challenged group at 37 or 46 months after ML inoculation, although the BCG-vaccinated group was significantly protected from leprosy and the unvaccinated group was not . In contrast, at 37 months the Fernandez reaction was positive in the BCG plus LD and the BCG plus HD HKML-vaccinated groups, both of which were significantly protected from clinical leprosy . By 46 months, the Fernandez responses were below significance in all groups . Thus, Fernandez reactivity is not a reliable correlate to protection from experimental leprosy in RM . Mitsuda responses became strongly positive in all four ML-challenged groups by 37 months and remained strongly positive at 46 months after ML inoculation, suggesting that strong Mitsuda reactivity reflects responses to living ML . BCG or BCG + LD or HD HKML vaccination/boosting of RM produced significant clinical protection from leprosy and there was a good correlation between protection from LL forms of leprosy and positive Mitsuda skin test responses after challenge with live ML . Positive Mitsuda responses were generated in essentially all individuals after challenge with live ML, and this response was primed by prior vaccination/boosting with BCG + HKML as shown by conversion to positivity 2 weeks after boosting . The data show that resistance to clinical leprosy is reflected by Mitsuda responses in ML-exposed RM, similar to results from human studies, and confirm the suitability of RM as a model for leprosy vaccine studies. Prikl Biokhim Mikrobiol, 2002 Nov-Dec, 38(6), 669 - 76 {Development of the biological preparation enatin with broad-range antimicrobial action}; Romanovskaia TV et al.; Physiological and biochemical traits of epiphytic spore forming bacteria Bacillus pumilis BIM V-263 were examined . The nutrient medium and conditions for submerged cultivation of the strain were selected . The growth dynamics and antagonistic activity during cultivation in a laboratory fermenter ANKUM-2M were studied . The results provide grounds for development of the biological preparation Enatin with broad-range antimicrobial effect . The plant-protective and growth-stimulating effect of Enatin was examined in laboratory and field experiments . The preparation holds promise as means for biological control of crop pathogens. Prikl Biokhim Mikrobiol, 2002 Nov-Dec, 38(6), 634 - 8 {Biosynthesis of inulinases by Bacillus bacteria}; Zherebtsov NA et al.; Biosynthesis of extracellular inulinase by bacteria Bacillus polymyxa 29, B . polymyxa 722, and B . subtilis 68 was studied . The optimal parameters for the producer growth were as follows: pH 7.0, 33-35 degrees C, the growth duration within 72 h . The presence of mineral reduced or of organic nitrogen was necessary for the enzyme biosynthesis . The inulinase biosynthesis was sharply activated in the presence of carbohydrates . B . polymyxa 722 and B . polymyxa 29 displayed the maximal activity on a starch-containing culture medium, the maximal activity of B . subtilis 68 was found in the presence of sucrose . Inulin did not induce the inulinase biosynthesis by the strains studied . The time course of bacteria growth and of the enzyme biosynthesis was studied. Antonie Van Leeuwenhoek, 2002 Aug, 81(1-4), 565 - 74 Toward an understanding of microbial communities through analysis of communication networks; Dunn AK et al.; Bacteria receive signals from diverse members of their biotic environment . They sense their own species through the process of quorum sensing, which detects the density of bacterial cells and regulates functions such as bioluminescence, virulence, and competence . Bacteria also respond to the presence of other microorganisms and eukaryotic hosts . Most studies of microbial communication focus on signaling between the microbe and one other organism for empirical simplicity and because few experimental systems offer the opportunity to study communication among various types of organisms . But in the real biological world, microorganisms must carry on multiple molecular conversations simultaneously between diverse organisms, thereby constructing communication networks . We propose that biocontrol of plant disease, the process of suppressing disease through application of a microorganism, offers a model for the study of communication among multiple organisms . Successful biocontrol requires the sending and receiving of signals between the biocontrol agent and the pathogen, plant host, and microbial community surrounding the host . We are using Bacillus cereus, a biocontrol agent, and the organisms it must interact with, to dissect a communication network . This system offers an excellent starting point for study because its members are defined and well studied . An understanding of signaling in the B . cereus biocontrol system may provide a model for network communication among organisms that share a habitat and provide a new angle of analysis for understanding the interconnections that define communities. Water Res, 2002 Nov, 36(19), 4850 - 60 Studies on the production of B . thuringiensis based biopesticides using wastewater sludge as a raw material; Vidyarthi AS et al.; Growth and delta-endotoxin yield of Bacillus thuringiensis (Bt) subsp kurstaki in tryptic soy yeast extract (TSY) medium, soybean meal based commercial medium and wastewater sludge medium were studied . The viable spores (VS) count in sludge medium was comparable to that obtained in laboratory and commercial media . The entomotoxicity of the fermentation liquid (Bt grown sludge) against Choristoneura fumiferana was comparable to the concentrated commercial Bt formulation available in the market (Foray 48B) . A higher entomotoxicity was observed in a sludge medium than in the TSY or soybean meal media . The secondary and mixed (mixture of primary and secondary) sludges from various wastewater treatment plants were also evaluated for spore formation and entomotoxicity yield . The VS count was higher in a mixed sludge compared to the secondary sludge at a similar sludge solids concentration . Both VS count and entomotoxicity yield was found to be a function of sludge solids concentration in the medium . The optimum value of solids concentration for Bt production was found to be 25 g (-1) (dry weight basis) . Beyond this concentration, a drop in VS count and entomotoxicity yield was observed . A low C:N ratio in the secondary sludge and a high C:N ratio in the mixed sludge resulted in a higher entomotoxicity . The optimum value of C:N ratio in combined sludge for Bt production was found to be 7.9-9.9 . Relationships between entomotoxicity and maximum specific growth as well as with specific sporulation rate were developed. J Infect Dis, 2002 Dec 15, 186(12), 1797 - 807 Epub 2002 Nov 19. Tuberculin skin testing and in vitro T cell responses to ESAT-6 and culture filtrate protein 10 after infection with Mycobacterium marinum or M . kansasii; Arend SM et al.; T cell responses to ESAT-6 and culture filtrate protein 10 (CFP-10), antigens expressed by Mycobacterium tuberculosis but not by M . bovis bacille Calmette-Guerin (BCG), were found to discriminate reliably between infection with M . tuberculosis and BCG vaccination . Because the esat-6 and cfp-10 genes occur in M . kansasii and M . marinum, T cell responses to ESAT-6 and CFP-10 were investigated in patients infected with M . kansasii or M . marinum, persons intensively exposed to environmental mycobacteria, and unexposed control subjects . Tuberculin skin tests were performed, and peripheral blood mononuclear cells were cocultured with ESAT-6, CFP-10, peptide mixtures of ESAT-6 and CFP-10, and control antigens . When enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot assay (ELISPOT) were used to measure interferon-gamma production, most M . kansasii- or M . marinum-infected patients and several persons exposed to environmental mycobacteria were found to respond to ESAT-6 and/or CFP-10 . ELISA and ELISPOT yielded comparable results, as did whole antigen and peptides (P<.0001) . These results may be relevant for the development of novel assays for diagnosis of tuberculosis. J Infect Dis, 2002 Dec 15, 186(12), 1733 - 9 Epub 2002 Nov 22. Molecular epidemiology of Bartonella henselae infection in human immunodeficiency virus-infected patients and their cat contacts, using pulsed-field gel electrophoresis and genotyping; Chang CC et al.; Bartonella henselae causes severe disease in immunocompromised individuals . B . henselae was isolated from 12 human immunodeficiency virus (HIV)-infected individuals with bacillary angiomatosis and/or peliosis hepatis and from their 15 cat contacts . Specific associations between the 2 B . henselae genotypes, individual pulsed-field gel electrophoresis (PFGE) patterns, and different clinical syndromes and pathogenicity were investigated . The role of cat contacts as the source of human infection was also examined . Three of the 4 patients with B . henselae genotype I infection, but none of the 8 patients with genotype II infection, had hepatosplenic vascular proliferative lesions (P=.018) . Four of 5 human-cat pairs had closely-related PFGE patterns and concordant results by 16S rDNA typing, which strongly suggests that human infection was caused by the cat contact . These results corroborate the major role of cats in the transmission of B . henselae to humans and suggest that B . henselae genotypes may induce different pathological features in HIV-infected patients. J Invertebr Pathol, 2002 Oct, 81(2), 122 - 6 Assessment of the efficacy of Japanese Bacillus thuringiensis isolates against the cigarette beetle, Lasioderma serricorne (Coleoptera: Anobiidae); Tsuchiya S et al.; A total of 2,652 Japanese isolates of Bacillus thuringiensis, belonging to at least 54 H serogroups, were examined for assessment of the toxicity against the cigarette beetle, Lasioderma serricorne (Coleoptera: Anobiidae) . When tested with spore/parasporal inclusion mixtures, strong larvicidal activities were associated with 28 isolates (1.1%) . Serologically, these toxic isolates fell into 4 known H serovars: thuringiensis (9 isolates), kurstaki (2), kenyae (2), and darmstadiensis (15) . Purified parasporal inclusions of the 10 selected isolates exhibited no larvicidal activity, while the supernatants of liquid cultures showed larvicidal and/or growth inhibitory effects . The activities were fully retained after heat treatment at 100 degrees C for 10 min . Overall results suggest that beta-exotoxin (or thuringiensin)-related substances are responsible for the toxicity of the present B . thuringiensis isolates against the cigarette beetle. FEMS Microbiol Lett, 2002 Nov 19, 217(1), 115 - 9 The enterotoxin T (BcET) from Bacillus cereus can probably not contribute to food poisoning; Choma C et al.; A polymerase chain reaction (PCR) fragment from strain NVH 38 (containing bceT) was cloned, sequenced and expressed in Escherichia coli . This sequence showed 50-60% identity to the original . When this bceT clone was expressed in E . coli no biological activity was found in either supernatants or cell extracts . Cell extracts from the Bacillus cereus strains (NVH 38 and B-4ac) were also negative on Vero cells . Neutralisation of supernatant from B . cereus B-4ac using a monoclonal antibody (reacting with NheB and HblD) abolished the activity . A test for cytotoxic enterotoxins was negative for both cell extracts and supernatants . Our data suggest that BcET either has an unknown type enterotoxic action or non at all. J Nat Prod, 2002 Nov, 65(11), 1664 - 6 Limnazine, the first bacterial