Bacillus

Lett Appl Microbiol, 2003, 36(5), 282 - 7
Physical properties of an extracellular polysaccharide produced by Bacillus sp . CP912; Yun UJ et al.; AIMS: In this study, some physical properties of Bacillus sp . exo-polysaccharide were investigated . METHODS AND RESULTS: An extracellular polysaccharide was purified by sequential precipitations after homogenization of the diluted culture supernatant of Bacillus sp . CP912 . Its physical properties were examined such as lipid emulsifying effect on several vegetable oils and flocculating activity against the activated carbon suspension . The melting point and endothermic calories of the polysaccharide were 128.7 degrees C and 50.864 kCal mol-1, respectively . Its pyrolysis temperature was 284.58 degrees C . The polysaccharide showed high lipid emulsifying activity on oil-water emulsion, against olive, peanut, sunflower and corn oils . It exhibited high flocculating activity as well against activated carbon . CONCLUSIONS: The present findings suggest that the extracellular polysaccharide produced by Bacillus sp . CP912 has a great industrial potential because of its high lipid emulsifying and flocculating activity . SIGNIFICANCE AND IMPACT OF THE STUDY: These data represent a novel Bacillus sp . extracellular polysaccharide possessing high emulsifying and flocculating effects.

J Environ Sci Health A Tox Hazard Subst Environ Eng, 2003 Mar, 38(3), 483 - 92
Production of biosurfactant and its role in the biodegradation of oil hydrocarbons; Lu XX et al.; Based on indigenous bacteria from the oil-contaminated site of Dawu water source area in Zibo city of China, the production of biosurfactant and its role in the biodegradation of oil hydrocarbons have been investigated . Batch experiments were performed with paraffin as the sole substrate under temperature of 30 degrees C and pH of 7 . Two high-effective species of bacteria (Z1 and Z2) were isolated . During the growth of both species, biosurfactants were produced, as indicated by the decrease of surface tensions of the enrichment medium from 61 to 45 mN/m . The critical micelle concentration (CMC) of Z1 ferment liquid was 0.4 (volume content) . When reaching the CMC, Z1 ferment liquid demonstrated the effect of dissolution enhancement, that is, it could enhance the dissolved concentration of phenanthrene to above 1 mg/L, which was higher than the saturated solubility of phenanthrenene under standard condition . Elementary identification results revealed that both Z1 and Z2 belongs to bacillus . Z1 was 0.5 microm in diameter, 0.5-3 microm in length, and bearing flagellum, while Z2 was 0.2 microm in diameter, 0.5-1 microm in length and no flagellum.

J Biomed Mater Res B Appl Biomater, 2003 May 15, 65(2), 239 - 44
Optimization of a RF-generated CF4/O2 gas plasma sterilization process; Lassen KS et al.; A sterilization process with the use of RF-generated (13.56 MHz) CF(4)/O(2) gas plasma was optimized in regards to power, flow rate, exposure time, and RF-system type . The dependency of the sporicidal effect on the spore inoculum positioning in the chamber of the RF systems was also investigated . Dried Bacillus stearothermophilus ATCC 7953 endospores were used as test organisms . The treatments were evaluated on the basis of survival curves and corresponding D values . The only parameter found to affect the sterilization process was the power of the RF system . Higher power resulted in higher kill . Finally, when the samples were placed more than 3-8 cm away from a centrally placed electrode in System 2, the sporicidal effect was reduced . The results are discussed and compared to results from the present literature . The RF excitation source is evaluated to be more appropriate for sterilization processes than the MW source .

J Urol, 2003 May, 169(5), 1706 - 8
Defining bacillus Calmette-Guerin refractory superficial bladder tumors; Herr HW et al.; PURPOSE: We define bacillus Calmette-Guerin (BCG) refractory, high risk, superficial bladder cancer . MATERIALS AND METHODS: A total of 93 patients received a 6-week induction course of BCG . They were evaluated for response after 3 and 6 months . Half of the patients received monthly maintenance BCG for 2 years and half did not . In both groups the initial responses to BCG at 3 and 6 months were correlated with subsequent tumor recurrence and progression . RESULTS: Of the 93 cases 57% were negative for tumor at 3 months and 43% had residual tumor resected . At 6 months 80% of the patients were tumor-free and 20% had persistent or recurrent tumor . Maintenance BCG did not decrease tumor recurrence further than induction BCG . Subsequent tumor-free interval during 24 months of followup were best predicted by response to BCG after 6 months . CONCLUSIONS: A minimum treatment and followup time of 6 months is required to identify high risk, superficial bladder tumors as truly BCG refractory.

J Urol, 2003 May, 169(5), 1702 - 5
The predictive value of purified protein derivative results on complications and prognosis in patients with bladder cancer treated with bacillus Calmette-Guerin; Bilen CY et al.; PURPOSE: We investigate the correlation of purified protein derivative (PPD) results before intravesical bacillus Calmette-Guerin (BCG) instillations with prognosis and complications of BCG . MATERIALS AND METHODS: A total of 57 men and 4 women with proven intermediate or high risk superficial bladder cancer received 6 courses of intravesical BCG instillations following complete resection of tumors . Skin reactivity to a PPD derivative of Mycobacterium tuberculosis was tested before starting and 1 week after BCG . The test was considered positive if the induration was 10 mm . or more in diameter after 48 or 72 hours . The patients were grouped according to PPD responses and symptoms . The statistical analyses were performed between PPD positive and negative groups, and also between symptomatic and asymptomatic patients . The groups were compared for relapse rates, time to first recurrence, complication rates and clinical outcome . RESULTS: Most of the patients with systemic side effects were in the PPD positive group but only fever had a statistically significant difference and was more frequent in the positive group (p <0.05) . The recurrence-free intervals after intravesical BCG therapy did not differ significantly between PPD positive and negative groups . However, the trend of longer recurrence-free survival was evident for symptomatic patients (p = 0.056) . The numbers of tumor recurrences were 10 (52%) in the PPD negative group and 19 (51%) in the PPD positive group, which was statistically insignificant . CONCLUSIONS: Patients with systemic reactions to BCG had the longest disease-free survival . It seems that patients with an augmented reaction to BCG probably have better antitumor activity . Furthermore, although larger groups of patients are mandatory for statistical analysis, this study shows that hypersensitivity reaction against tuberculin could alert physicians of severe complications.

Biochim Biophys Acta, 2003 Apr 11, 1647(1-2), 24 - 9
Structure-function relationship in serine hydroxymethyltransferase; Appaji Rao N et al.; Serine hydroxymethyltransferase (SHMT), a pyridoxal-5'-phosphate (PLP)-dependent enzyme catalyzes the tetrahydrofolate (H(4)-folate)-dependent retro-aldol cleavage of serine to form 5,10-methylene H(4)-folate and glycine . The structure-function relationship of SHMT was studied in our laboratory initially by mutation of residues that are conserved in all SHMTs and later by structure-based mutagenesis of residues located in the active site . The analysis of mutants showed that K71, Y72, R80, D89, W110, S202, C203, H304, H306 and H356 residues are involved in maintenance of the oligomeric structure . The mutation of D227, a residue involved in charge relay system, led to the formation of inactive dimers, indicating that this residue has a role in maintaining the tetrameric structure and catalysis . E74, a residue appropriately positioned in the structure of the enzyme to carry out proton abstraction, was shown by characterization of E74Q and E74K mutants to be involved in conversion of the enzyme from an 'open' to 'closed' conformation rather than proton abstraction from the hydroxyl group of serine . K256, the residue involved in the formation of Schiffs base with PLP, also plays a crucial role in the maintenance of the tetrameric structure . Mutation of R262 residue established the importance of distal interactions in facilitating catalysis and Y82 is not involved in the formaldehyde transfer via the postulated hemiacetal intermediate but plays a role in stabilizing the quinonoid intermediate . The mutational analysis of scSHMT along with the structure of recombinant Bacillus stearothermophilus SHMT and its substrate(s) complexes was used to provide evidence for a direct transfer mechanism rather than retro-aldol cleavage for the reaction catalyzed by SHMT.

J Plant Physiol, 2003 Feb, 160(2), 105 - 13
Systemic induction of the biosynthesis of terpenic compounds in Digitalis lanata; Gutierrez Manero FJ et al.; A bacterial screening was carried out in the rhizosphere of two Digitalis species, D . thapsi and D . parviflora, both at the vegetative stage and at flowering . A total of 480 isolates were characterised at genus level, Bacillus being the dominant genera in all cases . Fifty percent of the Bacillus strains isolated from each species were analysed by PCR-RAPDs . At 85% similarity, 12 groups separated for D . thapsi and 18 for D . parviflora . One strain of each group was selected for biological assay on D . lanata, evaluating growth promotion and cardenolide content in leaves after inoculation performed in the root system . The plant parameters evaluated were leaf surface area, shoot and root dry weight and leaf number . Lanatoside C content was evaluated by HPLC . Only 17 strains caused significant increases in at least one of the parameters evaluated . The most striking result was that some strains promoted growth and increased cardenolide content at the same time . This effect was detected on leaves while inoculation was carried out on roots . Interestingly, these two parameters are not enhanced simultaneously under regular conditions in pot or in tissue cultures.

Appl Microbiol Biotechnol, 2003 Oct, 62(5-6), 569 - 73 Epub 2003 Apr 03.
In vitro reduction of hexavalent chromium by a cell-free extract of Bacillus sp . ES 29 stimulated by Cu2+; Camargo FA et al.; Chromium-resistant bacteria (CRB) isolated from soils can be used to reduce toxic Cr(VI) from contaminated environments . This study assessed in vitro reduction of hexavalent Cr using a cell-free extract (CFE) of CRB isolated from soil contaminated with dichromate . One isolate, ES 29, that substantially reduced Cr(VI) was identified as a Bacillus species by 16S rRNA gene-sequence homology . The isolate reduced Cr(VI) under aerobic conditions, using NADH as an electron donor and produced a soluble Cr(VI)-reducing enzyme stimulated by copper (Cu2+) . The CFE of the bacterial isolate reduced 50% of Cr(VI) in 6 h . The Cr(VI)-reduction activity of the CFE had a Km of 7.09 microM and a Vmax of 0.171 micromol min(-1) mg(-1) protein . Mercury inhibited the enzyme, but not competitively, with a Vmax of 0.143 micromol min(-1) mg(-1) protein, a Km of 7.07 microM and a Ki of 1.58 microM . This study characterizes the enzymatic reduction of Cr(VI) by Bacillus sp . ES 29 which can be used for the bioremediation of chromate.

Plant Mol Biol, 2003 Mar, 51(5), 789 - 801
Microbial starch-binding domains as a tool for targeting proteins to granules during starch biosynthesis; Ji Q et al.; Modification of starch biosynthesis pathways holds an enormous potential for tailoring granules or polymers with new functionalities . In this study, we explored the possibility of engineering artificial granule-bound proteins, which can be incorporated in the granule during biosynthesis . The starch-binding domain (SBD)-encoding region of cyclodextrin glycosyltransferase from Bacillus circulans was fused to the sequence encoding the transit peptide (amyloplast entry) of potato granule-bound starch synthase I (GBSS I) . The synthetic gene was expressed in the tubers of two potato cultivars (cv . Kardal and cv . Karnico) and one amylose-free (amf) potato mutant . SBDs accumulated inside starch granules, not at the granule surface . Amylose-free granules contained 8 times more SBD (estimated at ca . 1.6% of dry weight) than the amylose-containing ones . No consistent differences in physicochemical properties between transgenic SBD starches and their corresponding controls were found, suggesting that SBD can be used as an anchor for effector proteins without having side-effects . To test this, a construct harbouring the GBSS I transit peptide, the luciferase reporter gene, a PT-linker, and the SBD (in frame), and a similar construct without the linker and the SBD, were introduced in cv . Kardal . The fusion protein accumulated in starch granules (with retainment of luciferase activity), whereas the luciferase alone did not . Our results demonstrate that SBD technology can be developed into a true platform technology, in which SBDs can be fused to a large choice of effector proteins to generate potato starches with new or improved functionalities.

Recenti Prog Med, 2003 Apr, 94(4), 177 - 85
{Bartonellosis}; Mansueto P et al.; In the last years the number of Bartonella species significantly raised, often with an epidemiological profile of emergent disease . B . bacilliformis is the etiological agent of Carrion's disease . B . clarridgeiae has been associated, together with B . henselae, to Cat-Scratch Disease (CSD), whereas B . elizabethae to endocarditis and B . grahamii to neuroretinitis . B . henselae has been associated to CSD and, in patients with immunodeficiency, together with B . quintana, to bacillary angiomatosis, to peliosis and to endocarditis . B . quintana is the etiological agent of "trench fever", in its "classical" and "urban" form . B . vinsonii subsp . arupensis and subsp . berkhoffii have been associated to bacteremia and endocarditis . The diagnosis foresees pathological examination of the lesions, germ isolation and use of specific serological test and of polymerase chain reaction . Prognosis is, generally, favourable . Therapy foresees the use of numerous antibacterial agents, like: penicillins, cephalosporins, aminoglycosides, tetracyclines, macrolides, quinolones, trimethoprim-sulfomethoxazole and rifampicin.

Arch Microbiol, 2003 May, 179(5), 363 - 7 Epub 2003 Apr 02.
Activity of wild-type and hybrid Bacillus thuringiensis delta-endotoxins against Agrotis ipsilon; de Maagd RA et al.; Twelve Cry1 and two Cry9 delta-endotoxins from Bacillus thuringiensis were tested for their activity against black cutworm ( Agrotis ipsilon) . A . ipsilon was not susceptible to many toxins, but three toxins had significant activity . Cry9Ca was the most toxic, followed by Cry1Aa and Cry1Fb . Hybrids between these three active proteins were made by in vivo recombination and analyzed for activity against A . ipsilon . Analysis of hybrids between Cry1Aa and Cry1Fb indicated that domain I of Cry1Aa protein was involved in its higher activity.

Appl Environ Microbiol, 2003 Apr, 69(4), 2245 - 52
Killing of bacillus spores by aqueous dissolved oxygen, ascorbic acid, and copper ions; Cross JB et al.; An approach to decontamination of biological endospores is discussed . Specifically, the performance of an aqueous modified Fenton reagent is examined . A modified Fenton reagent formulation of cupric chloride, ascorbic acid, and sodium chloride is shown to be an effective sporicide under aerobic conditions . The traditional Fenton reaction involves the conversion of hydrogen peroxide to hydroxyl radical by aqueous ionic catalysts such as the transition metal ions . Our modified Fenton reaction involves the conversion of aqueous dissolved oxygen to hydrogen peroxide by an ionic catalyst (Cu(2+)) and then subsequent conversion to hydroxyl radicals . Results are given for the modified Fenton reagent deactivating spores of Bacillus globigii . A biocidal mechanism is proposed that is consistent with our experimental results and independently derived information found in the literature . This mechanism requires diffusion of relatively benign species into the interior of the spore, where dissolved O(2) is then converted through a series of reactions which ultimately produce hydroxyl radicals that perform the killing action.

Appl Environ Microbiol, 2003 Apr, 69(4), 2065 - 72
Effects of inactivation and constitutive expression of the unfolded- protein response pathway on protein production in the yeast Saccharomyces cerevisiae; Valkonen M et al.; One strategy to obtain better yields of secreted proteins has been overexpression of single endoplasmic reticulum-resident foldases or chaperones . We report here that manipulation of the unfolded-protein response (UPR) pathway regulator, HAC1, affects production of both native and foreign proteins in the yeast Saccharomyces cerevisiae . The effects of HAC1 deletion and overexpression on the production of a native protein, invertase, and two foreign proteins, Bacillus amyloliquefaciens alpha-amylase and Trichoderma reesei endoglucanase EGI, were studied . Disruption of HAC1 caused decreases in the secretion of both alpha-amylase (70 to 75% reduction) and EGI (40 to 50% reduction) compared to the secretion by the parental strain . Constitutive overexpression of HAC1 caused a 70% increase in alpha-amylase secretion but had no effect on EGI secretion . The invertase levels were twofold higher in the strain overexpressing HAC1 . Also, the effect of the active form of T . reesei hac1 was tested in S . cerevisiae . hac1 expression caused a 2.4-fold increase in the secretion of alpha-amylase in S . cerevisiae and also slight increases in invertase and total protein production . Overexpression of both S . cerevisiae HAC1 and T . reesei hac1 caused an increase in the expression of the known UPR target gene KAR2 at early time points during cultivation.

Mol Ecol, 2003 Mar, 12(3), 765 - 75
Degradation of the Cry1Ab protein within transgenic Bacillus thuringiensis corn tissue in the field; Zwahlen C et al.; Large quantities of Bacillus thuringiensis (Bt) corn plant residue are left in the field after harvest, which may have implications for the soil ecosystem . Potential impacts on soil organisms will also depend on the persistence of the Bt toxin in plant residues . Therefore, it is important to know how long the toxin persists in plant residues . In two field studies in the temperate corn-growing region of Switzerland we investigated degradation of the Cry1Ab toxin in transgenic Bt corn leaves during autumn, winter and spring using an enzyme-linked immunosorbent assay (ELISA) . In the first field trial, representing a tillage system, no degradation of the Cry1Ab toxin was observed during the first month . During the second month, Cry1Ab toxin concentrations decreased to approximately 20% of their initial values . During winter, there was no further degradation . When temperatures again increased in spring, the toxin continued to degrade slowly, but could still be detected in June . In the second field trial, representing a no-tillage system, Cry1Ab toxin concentrations decreased without initial delay as for soil-incorporated Bt plants, to 38% of the initial concentration during the first 40 days . They then continued to decrease until the end of the trial after 200 days in June, when 0.3% of the initial amount of Cry1Ab toxin was detected . Our results suggest that extended pre- and post-commercial monitoring are necessary to assess the long-term impact of Bt toxin in transgenic plant residues on soil organisms.

Mol Microbiol, 2003 Apr, 48(2), 417 - 27
Multiplication of a restriction-modification gene complex; Sadykov M et al.; Previous works have suggested that some gene complexes encoding a restriction (R) enzyme and a cognate modification (M) enzyme may behave as selfish mobile genetic elements . RM gene complexes, which destroy 'non-self' elements marked by the absence of proper methylation, are often associated with mobile genetic elements and are involved in various genome rearrangements . Here, we found amplification of a restriction-modification gene complex . BamHI gene complex inserted into the Bacillus chromosome showed resistance to replacement by a homologous stretch of DNA . Some cells became transformed with the donor without losing BamHI . In most of these transformants, multiple copies of BamHI and the donor allele were arranged as tandem repeats . When a clone carrying one copy of each allele was propagated, extensive amplification of BamHI and the donor unit was observed in a manner dependent on restriction enzyme gene . This suggests that restriction cutting of the genome participates in the amplification . Visualization by fluorescent in situ hybridization revealed that the amplification occurred in single cells in a burst-like fashion that is reminiscent of induction of provirus replication . The multiplication ability in a bacterium with natural capacity for DNA release, uptake and transformation will be discussed in relation to spreading of RM gene -complexes.

Biotechnol Prog, 2003 Mar-Apr, 19(2), 652 - 4
Improved catalytic performance of Bacillus megaterium epoxide hydrolase in a medium containing Tween-80; Gong PF et al.; A new epoxide hydrolase with high enantioselectivity toward (R)-glycidyl phenyl ether (R-GPE) was partially purified from Bacillus megaterium strain ECU1001 . The maximum activity of the isolated enzyme was observed at 30 degrees C and pH 6.5 in a buffer system with 5% (v/v) of DMSO as a cosolvent . The enzyme was quite stable at pH 7.5 and retained full activity after incubation at 40 degrees C for 6 h . Interestingly, when the cosolvent DMSO was replaced by an emulsifier (Tween-80, 0.5% w/v) as an alternative additive to help disperse the water-insoluble substrate, the apparent activity of the epoxide hydrolase significantly increased by about 1.8-fold, while the temperature optimum shifted from 30 to 40 degrees C and the half-life of the enzyme at 50 degrees C increased by 2.5 times . The enzymatic hydrolysis of rac-GPE was highly enantioselective, with an E-value (enantiomeric ratio) of 69.3 in the Tween-80 emulsion system, which is obviously higher than that (41.2) observed in the DMSO-containing system.

Biotechnol Prog, 2003 Mar-Apr, 19(2), 493 - 4
Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) of alpha-amylases using a modified alginate; Mondal K et al.; The crude extracts of alpha-amylases when mixed with alginate, tert-butyl alcohol, and ammonium sulfate resulted in an interfacial precipitate containing polymer-bound amylase . The precipitate was dissolved in 1 M maltose to recover alpha-amylase activity . The recovery of alpha-amylases were 74%, 77%, and 92% in the case of Bacillus amyloliquefaciens, wheat germ, and porcine pancreas, respectively . All purified preparations showed a single band on SDS-PAGE.

Biol Chem, 2003 Feb, 384(2), 295 - 304
Primary structure and reactive site of a novel wheat proteinase inhibitor of subtilisin and chymotrypsin; Poerio E et al.; The proteinase inhibitor WSCI, active in inhibiting bacterial subtilisin and a number of animal chymotrypsins, was purified from endosperm of exaploid wheat (Triticum aestivum, c.v . San Pastore) by ion exchange chromatography and its complete amino acid sequence was established by automated Edman degradation . WSCI consists of a single polypeptide chain of 72 amino acid residues, has a molecular mass of 8126.3 Da and a pl of 5.8 . The inhibition constants (Ki) for Bacillus licheniformis subtilisin and bovine pancreatic alpha-chymotrypsin are 3.92 x 10(-9) M and 7.24 x 10(-9) M, respectively . The inhibitor contains one methionine and of tryptophan residue and has a high content of essential amino acids (41 over a total of 72 residues), but no cysteines . The primary structure of WSCI shows high similarity with barley subtilisin-chymotrypsin isoinhibitors of the Cl-2 type and with maize subtilisinchymotrypsin inhibitor MPI . Significant degrees of similarity were also found between sequences of WSCI and of other members of the potato inhibitor I family of the serine proteinase inhibitors . The wheat inhibitor WSCI has a single reactive site (the peptide bond between methionyl-48 and glutamyl-49 residues) as identified by affinity chromatography and sequence analysis.

Sheng Wu Gong Cheng Xue Bao, 2002 Nov, 18(6), 687 - 92
{Cloning and expression product of vip3A gene from Bacillus thuringiensis and analysis of inseceicidal activity}; Chen JW et al.; The vip3 A gene in a size of 2.3 kb amplified from wild-type Bacillus thuringiensis strain S184 by PCR was cloned into pGEM-T Easy vector and its sequence was analysized by DNASTAR . The plasmid pOTP was constructed by inserting vip3A-S184 gene into the expression vector pQE30 and then was transformed into E . coli M15 . E . coli M15 cells harbouring the plasmid pOTP were induced with 1 mmol/L IPTG to express 89 kD protein which was confirmed to be Vip3A-S184 by Western blot . Experiments showed that about 19% of Vip3A-S184 proteins were soluble, and others were insoluble proteins and formed inclusion bodies observed by transmission electron microscopy(TEM) . The target protein was purified under the native condition and the polyclonal antibody was prepared by immunizing rabbits . The polyclonal antibody was used to detect Vip3A proteins expressed in Bacillus thuringiensis . Bioassay showed that Vip3A-S184 showed a high toxicity against 3 tested insect larvae including Spodoptera exigua, Spodoptera litura and Helicoverpa armigera.

J Am Mosq Control Assoc, 2003 Mar, 19(1), 63 - 6
Ice granules containing endotoxins of microbial agents for the control of mosquito larvae--a new application technique; Becker N; This study investigated a new method for delivering microbial mosquito control agents into aquatic sites as ice granules for mosquito control . Solutions containing powder formulations of Bacillus thuringiensis israelensis (Bti) or Bacillus sphaericus were transformed into ice pellets (named IcyPearls) using a special ice-making machine . This new technique was demonstrated to have the following advantages over Bti sand granules: 1) the Bti ice pellets melted on the water surface and released the microbial crystals there; 2) the control agent remained inside the ice pellets during the application and were not lost by friction in the spraying equipment; and 3) the ice formulation resulted in increased swath widths, significantly reducing the cost of application . In large field tests . IcyPearls have been applied at dosages of 5 and 10 kg/ha containing 400 g as well as 100, 200, and 400 g of VectoBac WDG (3,000 ITU/mg), respectively, against larvae of Aedes vexans . Mortality rates of 91-98% were achieved.

J Am Mosq Control Assoc, 2003 Mar, 19(1), 39 - 46
Emergence of resistance and resistance management in field populations of tropical Culex quinquefasciatus to the microbial control agent Bacillus sphaericus; Mulla MS et al.; In recent years, highly potent mosquitocidal strains of the microbial agent Bacillus sphaericus (Bsph) have been isolated and developed for the control of mosquito larvae around the world . Laboratory selection experiments with the most active strains and their use in large-scale operational mosquito control programs resulted in the emergence of resistance in larvae of the Culex pipiens complex . This generated great concern among vector control agencies around the world, who feared reduced efficacy of this highly active larvicidal agent . To address this issue, the current studies were started to find practical strategies for controlling resistant mosquitoes and more importantly to develop resistance management strategies that would prevent or delay development of resistance . We initiated field studies in 3 low-income communities in Nonthaburi Province, Thailand . In 1 of the communities, larvae of Culex quinquefasciatus that were highly resistant (>125,000-fold) to Bsph strain 2362 were successfully controlled with applications of Bacillus thuringiensis var . israelensis (Bti) alone or in combination with Bsph . To prevent or delay resistance to Bsph, 2 other sites were selected, 1 treated with Bsph 2362 alone and the other treated with a mixture of Bsph 2362 and Bti . Mosquitoes treated with Bsph 2362 alone showed some resistance by the 9th treatment and almost complete failure of control occurred by the 17th treatment . After 9 treatments with the mixture over a 9-month period at another site, no noticeable change in susceptibility to Bsph was detected . During this period, the site treated with Bsph alone required 19 treatments, whereas the site treated with mixtures took only 9 treatments because of slower resurgence of larvae at the site treated with the mixture than at the site treated with Bsph alone . This is the 1st field evidence for delay or prevention of resistance to microbial agents in larval Cx . quinquefasciatus by using mixtures of Bti and Bsph . Further studies on the use of mixtures for the management of field resistance are warranted.

Can J Microbiol, 2003 Jan, 49(1), 37 - 44
Antibacterial activity of Cry- and Cyt-proteins from Bacillus thuringiensis ssp . israelensis; Yudina TG et al.; Mosquitocidal endotoxins Cry4B, Cry11A, and CytA from Bacillus thuringiensis ssp . israelensis as well as the products of their limited proteolysis display antibacterial activity relative to Micrococcus luteus . The endotoxin Cry11A also induces the lysis of the micrococcus protoplasts . Potassium and sodium ions and N-acetylgalactosamine increased the antibacterial effect of Cry11A, whereas glucose and N-acetylglucosamine inhibited it . The endotoxin Cry11A displays the antibacterial effect on some other microorganisms.

Cancer, 2003 Apr 15, 97(8 Suppl), 2099 - 108
Southwest Oncology Group studies in bladder cancer; Crawford ED et al.; Over 50,000 patients are diagnosed annually with bladder cancer, and approximately 10,000 eventually will die of their disease . Thus, the Southwest Oncology Group (SWOG) Genitourinary Cancer Committee is committed to the study of therapeutic interventions in patients with superficial, invasive, and metastatic bladder cancer . In the past 15 years, SWOG has completed six Phase III, randomized trials . Studies in patients with superficial disease have established the role of bacillus Calmette-Guerin in patient management; and a large, randomized trial has outlined the value of neoadjuvant chemotherapy and cystectomy in patients with advanced disease . SWOG plans to build on this model by evaluating patients with residual disease after chemotherapy for possible bladder preservation while evaluating more chemotherapy for patients with persistent disease . The Genitourinary Cancer Committee will continue to seek new, active agents for metastatic disease and will participate in and support large, Phase III, international trials that seek to improve current regimens . SWOG accomplishments in bladder cancer are highlighted, and future strategies are discussed .

Cancer, 2003 Apr 15, 97(8 Suppl), 2090 - 8
Overview of bladder cancer trials in the Cancer and Leukemia Group B; Small EJ et al.; The Cancer and Leukemia Group B (CALGB) Genitourinary Committee has developed a broad range of clinical trials across most stages of bladder cancer . Recurrence rates of superficial bladder cancer after transurethral resection range from 50-70% . Although adjuvant bacillus Calmette-Guerin reduces the risk of disease recurrence or progression, only 30% of patients have long-term disease-free survival . Because the development of novel secondline agents is needed, the CALGB is evaluating the utility of intravesicle gemcitabine as well as an oral proapoptotic agent (CP-461) . In patients with locally advanced disease with an increased risk of disease recurrence after cystectomy, a randomized trial of conventional chemotherapy versus sequential dose-dense therapy is under development . The gemcitabine/cisplatin combination has become a commonly used regimen for the treatment of advanced transitional cell carcinoma (TCC) . The CALGB is undertaking a Phase II study that incorporates a fixed dose rate gemcitabine infusion in this regimen, together with a selective epidermal growth factor receptor tyrosine kinase inhibitor, Iressa (Astra Zeneca, Wilmington, DE) . In patients with renal insufficiency, a regimen of carboplatin, gemcitabine, and Iressa is planned . Novel agents, including arsenic trioxide and trastuzumab (Herceptin; Genentech, Inc., South San Francisco, CA), are being evaluated as secondline therapy in patients with advanced TCC who have disease progression after frontline therapy .

Pediatr Res, 2003 Jul, 54(1), 105 - 12 Epub 2003 Apr 02.
BCG promotes cord blood monocyte-derived dendritic cell maturation with nuclear Rel-B up-regulation and cytosolic I kappa B alpha and beta degradation; Liu E et al.; Mycobacterium bovis bacillus Calmette-Guerin (BCG) is given to millions of neonates in developing countries as a vaccine against Mycobacterium tuberculosis; however, little is known about the initiation of response in neonatal dendritic cells (DCs) to BCG . To address this issue, the interaction of BCG with human cord blood monocyte-derived DCs was studied . We showed that BCG could promote cord blood monocyte-derived DC maturation by up-regulation of CD80, CD83, CD86, CD40, and MHC class II molecules and down-regulation of mannose receptor . BCG was able to induce similar levels of tumor necrosis factor-alpha and IL-10 but no bioactive IL-12p70 production from cord blood DCs as from adult blood DCs . Functionally BCG-treated cord blood DCs had higher ability to induce mixed lymphocyte reaction than non-BCG-treated cord blood DCs . Both non-BCG-treated and BCG-treated cord blood DCs efficiently induced a high level of IL-10, medium level of interferon-gamma, but little IL-4 production by cord blood naive CD4+ T cells . Heat shock protein 65, a key component of BCG, had no effect on cord blood DC maturation in terms of CD86, MHC class II, and mannose receptor up-regulation . During the BCG-induced maturation process of cord blood DCs, nuclear transcription factor Rel-B was up-regulated and cytosolic Rel-B down-regulated with cytosolic IkappaB alpha and beta degradation . These results suggest that BCG can promote cord blood monocyte-derived DC maturation, and that the mechanism is through the up-regulation of nuclear Rel-B secondary to the degradation of cytosolic IkappaB alpha and beta.

Acta Pol Pharm, 2002 Nov-Dec, 59(6), 433 - 5
Sterility and antibacterial activity of some antibiotics sterilized by irradiation; Muszynski Z et al.; Sterility and antibacterial activity of several antibiotics (including some penicillins and their salts, gramicidin and neomycin) subjected to sterilization by irradiation has been studied . The compounds in solid phase have been exposed to gamma irradiation in air atmosphere at room temperature, with a dose of 25 kGy, and afterwards they have been subjected to tests recommended by FP V (volume I, 1990) checking their sterility and activity . The results have shown that the majority of initial compounds have been to a slight degree contaminated by bacteria from the genera Bacillus and Micrococcus, the number of bacteria did not exceed 10(2) CFU, and fungi up to 10 CFU in 1 g of the compound . All compounds subjected to sterilization with a dose of 25 kGy were sterile and preserved the activity required by FP V . The decrease in activity observed for some compounds was always within the limits of FP specification . The results have proved that the penicillins analysed, gramicidin and neomycin can be sterilized by irradiation with a dose of 25 kGy, without any detrimental effect on their properties and antibacterial activity.

J Biol Chem, 2003 Jun 6, 278(23), 20659 - 66 Epub 2003 Mar 31.
Coordination geometries of metal ions in d- or l-captopril-inhibited metallo-beta-lactamases; Heinz U et al.; d- and l-captopril are competitive inhibitors of metallo-beta-lactamases . For the enzymes from Bacillus cereus (BcII) and Aeromonas hydrophila (CphA), we found that the mononuclear enzymes are the favored targets for inhibition . By combining results from extended x-ray absorption fine structure, perturbed angular correlation of gamma-rays spectroscopy, and a study of metal ion binding, we derived that for Cd(II)1-BcII, the thiolate sulfur of d-captopril binds to the metal ion located at the site defined by three histidine ligand residues . This is also the case for the inhibited Co(II)1 and Co(II)2 enzymes as observed by UV-visible spectroscopy . Although the single metal ion in Cd(II)1-BcII is distributed between both available binding sites in both the uninhibited and the inhibited enzyme, Cd(II)1-CphA shows only one defined ligand geometry with the thiolate sulfur coordinating to the metal ion in the site composed of 1 Cys, 1 His, and 1 Asp . CphA shows a strong preference for d-captopril, which is also reflected in a very rigid structure of the complex as determined by perturbed angular correlation spectroscopy . For BcII and CphA, which are representatives of the metallo-beta-lactamase subclasses B1 and B2, we find two different inhibitor binding modes.

J Virol Methods, 2003 Apr, 109(1), 89 - 93
Detection of molecular variability in rice tungro bacilliform viruses from India using polymerase chain reaction-restriction fragment length polymorphism; Joshi R et al.; Rice tungro bacilliform virus (RTBV) with rice tungro spherical virus (RTSV) causes the destructive tungro disease of rice . In order to ascertain the molecular variability of RTBV in India, primers were designed to amplify a polymorphic DNA fragment of the virus . Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis on a number of field isolates indicated mixed infections and molecular heterogeneity in the viral genome.

Microb Pathog, 2003 Apr, 34(4), 179 - 86
Differential expression of the invasion-associated locus B (ialB) gene of Bartonella bacilliformis in response to environmental cues; Coleman SA et al.; Bartonella bacilliformis is the causative agent of the biphasic human disease, Oroya fever . During the primary disease phase, up to 100% of the circulating erythrocytes can be parasitized and 80% lysed . During the secondary phase of this disease, bacterial invasion shifts to endothelial cells lining the vasculature . B . bacilliformis is transferred between human hosts by the sandfly, Lutzomyia verrucarum . To investigate the regulation of ialB by environmental cues signaling vector-to-host transmission; nuclease protection assays were performed to compare the amount of ialB mRNA in bacteria subjected to temperature shift, pH change, oxidative stress, or hemin limitation . The amount of ialB mRNA increased by 223-310% in acid-treated samples and decreased by 28-39% in base-treated samples as compared to bacteria kept at pH 7.2 . B . bacilliformis samples showed a 56-63% and 74-80% decrease in ialB mRNA when shifted to 37 degrees C from growth temperatures of 20 and 30 degrees C, respectively . Oxidative stress (1 mM H(2)O(2)) and hemin limitation had no significant effect on mRNA levels . Determination of IalB protein amounts using SDS-PAGE and immunoblotting showed the greatest amounts of IalB under acidic conditions or at 20 degrees C . The least amount of IalB was synthesized under basic conditions or at 37 degrees C . The viability of wild-type B . bacilliformis under the various experimental culture conditions was determined and found not to affect ialB mRNA amounts in these experiments . Finally, we compared the survival of wild-type and ialB mutant B . bacilliformis and found no difference in the viability of these two strains, demonstrating that IalB does not aid bacterial survival under these conditions .

Curr Opin Plant Biol, 2003 Apr, 6(2), 191 - 8
Advances in plant biotechnology and its adoption in developing countries; Toenniessen GH et al.; Developing countries are already benefiting and should continue to benefit significantly from advances in plant biotechnology . Insect-protected cotton containing a natural insecticide protein from Bacillus thuringiensis (Bt cotton) is providing millions of farmers with increased yields, reduced insecticide costs and fewer health risks . Many other useful plant biotechnology products that can benefit poor farmers and consumers are in the research and development pipelines of institutions in developing countries, and should soon reach farmers' fields.

Eur Urol, 2003 Apr, 43(4), 351 - 60; discussion 360-1
Do prognostic parameters of remission versus relapse after Bacillus Calmette-Guérin (BCG) immunotherapy exist? . analysis of a quarter century of literature; Saint F et al.; OBJECTIVE: To review prognostic factors identified in clinical trials for remission versus relapse after intravesical adjuvant Bacillus Calmette-Guerin (BCG) immunotherapy for superficial bladder cancer (Ta, T1, and carcinoma in situ) . MATERIALS AND METHODS: Information was retrieved by a MEDLINE search of the English literature . Indexing terms comprised bladder cancer, bladder neoplasm, BCG vaccine, superficial bladder cancer, immunotherapy, intravesical therapy, prognostic marker, and Bacillus Calmette-Guerin . Fifty clinical studies were assessed for the strength of their results on the therapeutic response to BCG instillation . Emphasis was placed on clinical trials that assessed tumor and/or host characteristics, immunological reactions, recurrence rates, progression rates and disease-specific survival after BCG . RESULTS: The predictive value of host factors is extremely controversial, but marked adverse reactions to BCG instillation appear to be associated with a better tumor response . Traditional pathological tumor characteristics, molecular markers (p53) and immunological status (PPD skin test) do not appear to have prognostic value in this setting . There is increasing evidence that immunologic markers are predictive of the BCG response, but most of them have not yet been assessed in large prospective studies . Histologic/cytologic response criteria are the critical determinant of post-BCG outcome . CONCLUSIONS: After a quarter century of clinical research, no independent prognostic factor for the bladder tumor response to BCG has yet been identified . Sophisticated individual therapeutic approaches (SITA) appear to be the most promising . Nomograms based on host, tumor and immunological characteristics may help with clinical decision-making and with rationalized BCG schedule design.

Biochim Biophys Acta, 2003 Apr 7, 1621(1), 31 - 40
A single surface tryptophan in the chitin-binding domain from Bacillus circulans chitinase A1 plays a pivotal role in binding chitin and can be modified to create an elutable affinity tag; Ferrandon S et al.; Site-directed mutagenesis was carried out to investigate the roles of a number of highly conserved residues of the chitin-binding domain (ChBD) of Bacillus circulans chitinase A1 (ChiA1) in the binding of chitin . Analysis of single alanine replacement mutants showed that mutation of an exposed tryptophan residue (Trp(687)) impaired the binding to chitin, while mutation of other highly conserved residues, most carrying aromatic or hydrophobic side chains, did not significantly affect the binding activity . Interestingly, replacement of Trp(687) with phenylalanine significantly reduced chitin-binding activity at lower salt concentrations (0-1 M NaCl) but allowed strong binding to chitin at 2 M NaCl . Since Trp(687) is conserved among the ChBDs belonging to the bacterial ChiA1 subfamily, the data presented suggest a general mechanism in which this exposed tryptophan, which is located in the cleft formed between two beta-sheets as revealed by the solution structure {J . Biol . Chem . 275 (2000) 13654}, makes a major contribution to ligand binding presumably through hydrophobic interactions . Furthermore, modulation of the chitin-binding activity by the conserved amino acid replacement (W687F) and a shift in the ionic strength of buffer has led to the development of an elutable affinity tag for single column purification of recombinant proteins.

Immunology, 2003 Apr, 108(4), 548 - 55
A DNA prime-live vaccine boost strategy in mice can augment IFN-gamma responses to mycobacterial antigens but does not increase the protective efficacy of two attenuated strains of Mycobacterium bovis against bovine tuberculosis; Skinner MA et al.; The Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccine has variable efficacy for both human and bovine tuberculosis . There is a need for improved vaccines or vaccine strategies for control of these diseases . A recently developed prime-boost strategy was investigated for vaccination against M . bovis infection in mice . BALB/c and C57BL/6 mice were primed with a DNA vaccine, expressing two mycobacterial antigens, ESAT-6 and antigen 85 A and boosted with attenuated M . bovis strains, BCG or WAg520, a newly attenuated strain, prior to aerosol challenge . Before challenge, the antigen-specific production of interferon-gamma (IFN-gamma) was evaluated by ELISPOT and antibody responses were measured . The prime-boost stimulated an increase in the numbers of IFN-gamma producing cells compared with DNA or live vaccination alone, but this varied according to the attenuated vaccine strain, time of challenge and the strain of mouse used . Animals vaccinated with DNA alone generated the strongest antibody response to mycobacterial antigens, which was predominantly IgG1 . BCG and WAg520 alone generally gave a 1-2 log10 reduction in bacterial load in lungs or spleen, compared to non-vaccinated or plasmid DNA only control groups . The prime-boost regimen was not more effective than BCG or WAg520 alone . These observations demonstrate the comparable efficacy of BCG and WAg520 in a mouse model of bovine tuberculosis . However, priming with the DNA vaccine and boosting with an attenuated M . bovis vaccine enhanced IFN-gamma immune responses compared to vaccinating with an attenuated M . bovis vaccine alone, but did not increase protection against a virulent M . bovis infection.

Przegl Epidemiol, 2002, 56(4), 577 - 86
{Eradication of leprosy and public health . Vaccination and multidrug therapy}; Janaszek W; Leprosy is a disease, which still affects large populations in the developing countries particularly in Africa, Asia and Latin America . For the last 15 years significant advances have been made towards leprosy elimination . The most effective strategy for leprosy control is an early identification of cases and an effective treatment with multidrug therapy (MDT) . The vaccination against leprosy plays only an additional role . There are two possible approaches to develop vaccine against leprosy . One is to produce a vaccine based on organisms related to M . leprae, such as: BCG, ICRC bacillus, Mycobacterium w, Mycobacterium vaccae, Mycobacterium habana . However, these organisms related to M . leprae are not very promising in experimental animal studies . In 1970s a new vaccine was prepared based on killed M . leprae . This vaccine, tested alone and together with BCG revealed little impact on increasing vaccine efficacy . The success in cloning and expressing the M . leprae genome in E . coli created the possibility of moving towards a second generation vaccine using peptide antigens . Up till now only MDT has essential impact on decline of global leprosy prevalence . Out of 122 endemic countries in 1985, 107 countries have reached elimination of leprosy at country level . At the end of 2000 leprosy was a public health problem only in 15 countries (prevalence rate > 1/10.000) . Currently leprosy remains a problem mainly in 6 major endemic countries . Among these, India alone accounts for 64% of prevalence and 78% of detection worldwide.

Biopolymers, 2003 Apr, 68(4), 486 - 96
Elongational flow studies on conformational change in DNA induced by DNA-binding protein HU; Endoh T et al.; Interaction of DNA-binding protein HU from Bacillus stearothermophilis (HUBst) with coliphage T2 DNA was investigated by observing an elongational flow-induced birefringence, Deltan, of a T2-phage DNA aqueous solution at various HU concentrations . Localized flow birefringence was observed in the pure elongational flow region, and the strain rate dependence of Deltan had a critical strain rate epsilon;(c) for the appearance of flow birefringence at all of the HU concentrations examined, indicating that a coil-stretch transition occurred at epsilon;(c) in each DNA-HU system . For strain rates larger than epsilon;(c), Deltan increased rapidly and then gradually, approaching a plateau value . The value of epsilon;(c) increased with an increase in HU concentration . Analysis based on the relationship between epsilon; (c) and the Rouse-Zimm relaxation time revealed that the increase in epsilon;(c)with increase in HU can be explained by the decrease in the size of the DNA-HU complex . The plateau birefringence value, Deltan(p), decreased at small HU concentrations but did not change at larger HU concentrations . Considering that Deltan(p) is related to the orientational order parameter of segments, it was concluded that there were at least two stages in the process of compaction of DNA induced by HU .

West Afr J Med, 2002 Oct-Dec, 21(4), 310 - 2
Neonatal intensive care unit: reservoirs of nosocomial pathogens; Newman MJ; Improvement in the care and treatment of neonates had contributed to their increased survival . Nosocomial infection remains an important problem in intensive care units . Hospital wards had been shown to act as reservoirs of pathogenic microorganisms associated with infection . To assess the prevalence of pathogenic organisms in the environment of the neonatal unit, 92 swabs were randomly collected from cots, incubators and various equipments in the unit and were cultured on Blood agar and MacConkey agar plates . Air contamination was detected by exposing the same types of agar plates for 3 hours in several areas of the unit . After 48 hours incubation, isolates were identified biochemically . There is marked congestion in the unit . Ninety one percent of swabs yielded growth, with coagulase negative Staphylococcus being the predominant organism (44%), followed by Bacillus species (20%), E . coli (12.5%), and Klebsiella (8.5%), Pseudomonas species (3.6%) and moulds (3.6%) . Sedimentation plates had colony counts of from 10 - 100 per plate and the majority of the cultures were polymicrobial cultures . The presence of various Gram-negative bacili including known neonatal pathogens (like E . Coli and Pseudomonas) especially on ward equipment and congestion in the ward has the potential to cause nosocomial infection.

Life Sci Space Res, 1970, 8, 77 - 81
The effect of high vacuum on oxidative reactions in bacteria and the activity of certain enzymes; Imshenetsky AA et al.; Among the physical factors which might influence micro-organisms one of the most potentially interesting is high vacuum . The effect of high vacuum is less studied as compared with other physical factors . It is impossible to achieve, under laboratory conditions, a vacuum of the order 10(-16) mm Hg which is probably characteristic of space . Earlier, the effect of high vacuum was studied on different bacteria, yeasts, molds and algae . It appeared that spores and fungal conidia were not killed by high vacuum . Later, the effect of high vacuum on physiological processes in micro-organisms was studied . The ability to oxidize glucose or ethanol was studied with Sarcina flava and Bacillus simplex cells after they were subjected for 72 hr to vacuum (10(-8) to 10(-9) mm Hg) . The oxidation rate was followed polarographically . The oxidative ability of S . flava cells diminished {correction of dimished} after their subjection to vacuum, while B . simplex spores were unchanged in that respect . The following crystalline enzymes were subjected for 72 hr to the same vacuum: alpha-amylase, catalase, ribonuclease, trypsine and urease . Then the activity of the above enzymes was tested on corresponding substrates . Not a single enzyme was totally inactivated . About 50% of activity was lost with alpha-amylase; 25--35% of activity with catalase, ribonuclease and urease . Trypsine retained its total activity . Thus, high vacuum cannot be listed among factors rapidly inactivating enzymes of micro-organisms.

J Clin Pathol, 2003 Apr, 56(4), 316 - 8
Anaerobiospirillum succiniciproducens bacteraemia; Pienaar C et al.; This report describes a case of bacteraemia caused by Anaerobiospirillum succiniciproducens . Anaerobiospirillum succiniciproducens is a rare cause of bacteraemia in humans, and when encountered usually occurs in immunocompromised patients . The organism is an anaerobic, spiral shaped, Gram negative bacillus with bipolar tufts of flagella . In this report, the morphology, with special reference to electron microscopic features, culture characteristics, and antimicrobial susceptibility are described.

J Zhejiang Univ Sci, 2003 Mar-Apr, 4(2), 142 - 51
Influence of medium components on elastase production using crude sources by Bacillus sp . EL31410; He GQ et al.; A newly isolated strain EL31410, producing elastase (E.C3.4.4.7) with high elastolytic activity was identified as Bacillus sp . In the medium optimization, it was found that wheat bran and soybean flour hydrosate were the best crude carbon and nitrogen source for enzyme production, respectively . Addition of corn steep flour can affect the bacterium growth and elastase production . A fractional factorial design was applied to study the main factors that affect the enzyme production, and central composite experimental design and response surface methodology were adopted to derive a statistical model for the effect of wheat bran and soybean flour hydrosate on elastase production . The experimental results showed that wheat bran had positive effect but soybean flour hydrosate had negative effect, on enzyme production . An initial concentration of 3.4% (w/v) wheat bran and 9.4% (v/v) soybean flour hydrosate were found to be optimal for enzyme production in batch culture . The time course of elastase production in the optimized medium composition was also described.

N Z Med J . 2003 Mar 14;116(1170):U354.
Symptom complaints following aerial spraying with biological insecticide Foray 48B; Petrie K et al.; AIM: To investigate the effect of aerial Bacillus thuringiensis (Foray 48B) spraying on self-reported symptom complaints, health perceptions, and visits to healthcare providers . METHODS: Two hundred and ninety two residents within the Ministry of Agriculture and Forestry (MAF) West Auckland spray zone were recruited by a door-to-door survey of homes within the most intensively sprayed area ten weeks prior to the first aerial spraying . Participants completed a symptom checklist and a questionnaire measuring health perceptions . Three months after the start of spraying, 181 (62%) of the original participants responded to a similar postal questionnaire . Symptom reports, health perceptions and visits to healthcare providers were compared between the baseline and the follow-up questionnaire . Rates of symptom complaints in respondents with previously diagnosed asthma, hay fever, or other allergies were compared to those in respondents without these prior health conditions . RESULTS: Symptom complaints increased significantly following the aerial spraying, in particular: sleep problems, dizziness, difficulty concentrating, irritated throat, itchy nose, diarrhoea, stomach discomfort, and gas discomfort . Analyses showed a significant increase in symptoms in those participants with a previous history of hay fever . While overall self-ratings of health decreased following the spraying, most residents saw their health as unaffected by the spray programme, and there was no significant increase in visits to general practitioners or alternative healthcare providers . CONCLUSIONS: Aerial spraying with Foray 48B is associated with some adverse health consequences in terms of significant increases in upper airway, gastrointestinal, and neuropsychiatric symptoms, as well as a reduction in overall perception of health in the exposed population./

Int J Urol, 2003 Apr, 10(4), 183 - 9
Sufficient prophylactic efficacy with minor adverse effects by intravesical instillation of low-dose bacillus Calmette-Guérin for superficial bladder cancer recurrence; Irie A et al.; BACKGROUND: Intravesical instillation of bacillus Calmette-Guerin (BCG) is the most efficient strategy for prophylaxis of superficial bladder cancer recurrence . Adverse effects of BCG are major obstacles, but the reduction of BCG dose could minimize these effects . The efficacy and adverse effects of half-dose (40 mg) BCG, Tokyo 172 strain, were prospectively evaluated . METHODS: A total of 93 patients with superficial bladder cancer (pTa or pT1) were sequentially assigned to receive either 40 or 80 mg of BCG after transurethral resection . BCG was administered weekly for 6 weeks postoperatively . Eighty patients observed longer than 12 months after BCG therapy (41, 40 mg group; 39, 80 mg group) were analyzed . RESULTS: BCG therapy course was completed in 71 patients . Tumor recurrence was recognized in 11 of 40 patients in the 40 mg group and in 5 of 31 patients in the 80 mg group . There was no significant difference in tumor recurrence rate between the two groups (P = 0.547) . BCG therapy was withdrawn in 1 patient in the 40 mg group and in 8 patients in the 80 mg-group because of BCG-related adverse effects . The morbidity of BCG-related toxicity was significantly higher in the 80 mg group . CONCLUSION: Half-dose of BCG Tokyo 172 strain had a similar efficacy and its toxicity was significantly lower compared to the standard dose . Thus, half-dose of this strain might be suitable, at least for initial BCG therapy, for the prophylaxis of bladder cancer recurrence . Further study would be necessary to clarify the efficacy of low-dose instillation in high-risk patients.

Mol Microbiol, 2003 Apr, 48(1), 77 - 84
Genes required for mycobacterial growth defined by high density mutagenesis; Sassetti CM et al.; Despite over a century of research, tuberculosis remains a leading cause of infectious death worldwide . Faced with increasing rates of drug resistance, the identification of genes that are required for the growth of this organism should provide new targets for the design of antimycobacterial agents . Here, we describe the use of transposon site hybridization (TraSH) to comprehensively identify the genes required by the causative agent, Mycobacterium tuberculosis, for optimal growth . These genes include those that can be assigned to essential pathways as well as many of unknown function . The genes important for the growth of M . tuberculosis are largely conserved in the degenerate genome of the leprosy bacillus, Mycobacterium leprae, indicating that non-essential functions have been selectively lost since this bacterium diverged from other mycobacteria . In contrast, a surprisingly high proportion of these genes lack identifiable orthologues in other bacteria, suggesting that the minimal gene set required for survival varies greatly between organisms with different evolutionary histories.

Kekkaku, 2003 Jan, 78(1), 45 - 9
{HIV infection and tuberculosis}; Nagai H; The number of people infected with human immunodeficiency virus (HIV) is gradually increasing in Japan, and the morbidity rate from tuberculosis in the Japanese people is high . Accordingly, the number of cases with both infections is considered to increase in the future . Our hospital has already encountered 31 cases of HIV associated tuberculosis . HIV infects mainly CD4-positive cells . The extreme decrease in the cell count results in serious cellular immunological disorder . CD4-positive cell disorder induces disorders of B lymphocytes, cytotoxic T cells, natural killer cells, and macrophage functions . These destructive conditions show the state of immunodeficiency including macrophage that are most important for defense of acid-fast bacterial infection . Migration and activation of macrophages with cytokines derived from T cells are impaired to induce the following phenomena: hypoplasia of granuloma, failure of tubercule bacillus suppression, the spread to regional lymph nodes (hilar or mediastinal lymph nodes), and hematogenous dissemination . On this occasion, caseous necrosis and cavitation are unlikely to occur, and false-negative tuberculin reaction is often observed . The incidence of severe cases, which include miliary tuberculosis, tuberculous meningitis, etc., and extrapulmonary tuberculosis, are high among acquired immunodeficiency syndrome (AIDS)-associated tuberculosis cases . HIV-infected tuberculosis cases are generally regarded as endogenous exacerbation, but they include primary infection and reinfection as well . Even during the treatment for drug-sensitive strains particularly, some cases may have reinfection with multidrug-resistant bacteria, suggesting that caution should be taken against this point . Conversely, the association of tuberculosis is a factor for the poor prognosis of HIV infection, since it facilitates the development of HIV infection . If the bacteria belong to a drug-sensitive strain, the infection with them responds well to antituberculous drugs, the same as in tuberculosis cases without HIV infection, showing a favorable prognosis . However, the mortality rate of infection with multi-drug-resistant tuberculosis is extremely high . The combined use of a protease inhibitor, i.e., anti-HIV drug, with rifampicin is regarded as contraindication for the treatment because rifampicin strongly induces hepatic cytochrome P-450 and increases the metabolism of protease inhibitors and nonnucleoside reverse transcriptases to markedly decrease the blood concentrations . Accordingly, the treatment for tuberculosis should take priority over that for HIV infection in HIV-infected tuberculosis, and highly active antiretroviral therapy (HAART) may be administered after the treatment of tuberculosis . When HAART is necessary for the treatment during the tuberculosis treatment, rifampicin had better be exchanged to rifabutin because the effect of rifabutin to induce cytochrome P-450 is less potent than that of rifampicin . A report has recently shown that the exacerbation of pyrexia and chest X-ray findings was transiently observed approximately 2 weeks after potent anti-HIV therapy for HIV-infected tuberculosis, which included a protease inhibitor . The reason for the exacerbation has been believed to be that the impaired function of CD4-positive cells is improved by the administration of anti-HIV drugs to raise temporarily the reaction of the vital part to M . tuberculosis . A tuberculin skin test (TST) reaction size of > or = 5 mm of induration is considered positive (i.e., indicative of M . tuberculosis infection) in persons who are infected with HIV . Persons with a TST reaction size > or = 5 mm who have not previously received treatment for M . tuberculosis infection should receive tuberculosis preventive treatment . Prevention by BCG vaccination is regarded as contraindications for HIV-infected patients, because disseminated M . bovis infection may be associated with them . Many HIV-positive patients infected with tuberculosis show uneventful healing, when M . tuberculosis is the sensitive strain . However, since some patients show the rapid course of tuberculosis, clinical physicians keep the early detection of tuberculosis for HIV-infected patients and the association of HIV infection for tuberculosis patients in their mind, respectively.

J Vet Med Sci, 2003 Feb, 65(2), 187 - 91
Effects of Bacillus thuringiensis Cry1Ab toxin on mammalian cells; Shimada N et al.; The Cry proteins produced by Bacillus thuringiensis are considered to be highly specific insecticidal proteins . Judged to be safe for humans and farm animals due to their insect-oriented selective toxicity, the proteins have been utilized as a biological pesticide and introduced into genetically modified plants . However, some critical fundamental characters of the Cry proteins remain unclear, and the direct effects of activated Cry proteins on mammalian cells have not yet been fully confirmed . Therefore, in this study we employed primary cultured bovine hapatocytes as a model system to determine if Cry1Ab, a Cry protein, affects mammalian cells . There were no significant changes in the secretion of albumin or the morphology of the Cry1Ab-treated cells . The LDH release showed a tendency to increase after the administration of Cry1Ab, but not significantly . Taking these results on bovine hepatocytes into consideration, Cry1Ab has little acute toxicity on mammalian cells.

Infect Immun, 2003 Apr, 71(4), 1763 - 73
Differential effects of control and antigen-specific T cells on intracellular mycobacterial growth; Worku S et al.; We investigated the effects of peripheral blood mononuclear cells expanded with irrelevant control and mycobacterial antigens on the intracellular growth of Mycobacterium bovis bacillus Calmette-Guerin (BCG) in human macrophages . More than 90% of the cells present after 1 week of in vitro expansion were CD3(+) . T cells were expanded from purified protein derivative-negative controls, persons with latent tuberculosis, and BCG-vaccinated individuals . T cells expanded with nonmycobacterial antigens enhanced the intracellular growth of BCG in suboptimal cultures of macrophages . T cells expanded with live BCG or lysates of Mycobacterium tuberculosis directly inhibited intracellular BCG . Recent intradermal BCG vaccination significantly enhanced the inhibitory activity of T cells expanded with mycobacterial antigens (P < 0.02), consistent with the induction of memory-immune inhibitory T-cell responses . Selected mycobacterial antigens (Mtb41 > lipoarabinomannan > 38kd > Ag85B > Mtb39) expanded inhibitory T cells, demonstrating the involvement of antigen-specific T cells in intracellular BCG inhibition . We studied the T-cell subsets and molecular mechanisms involved in the memory-immune inhibition of intracellular BCG . Mycobacteria-specific gammadelta T cells were the most potent inhibitors of intracellular BCG growth . Direct contact between T cells and macrophages was necessary for the BCG growth-enhancing and inhibitory activities mediated by control and mycobacteria-specific T cells, respectively . Increases in tumor necrosis factor alpha, interleukin-6, transforming growth factor beta, and vascular endothelial growth factor mRNA expression were associated with the enhancement of intracellular BCG growth . Increases in gamma interferon, FAS, FAS ligand, perforin, granzyme, and granulysin mRNA expression were associated with intracellular BCG inhibition . These culture systems provide in vitro models for studying the opposing T-cell mechanisms involved in mycobacterial survival and protective host immunity.

Infect Immun, 2003 Apr, 71(4), 1656 - 61
Virulence, immunogenicity, and protective efficacy of two recombinant Mycobacterium bovis bacillus Calmette-Guérin strains expressing the antigen ESAT-6 from Mycobacterium tuberculosis; Bao L et al.; We constructed two recombinant Mycobacterium bovis BCG (rBCG) strains expressing ESAT-6 of Mycobacterium tuberculosis, named rBCG-1 and rBCG-2 . rBCG-1 contained the ESAT-6 gene linked to BCG hsp60 and expressed a fusion protein, while rBCG-2, with a secretory sequence, could secret ESAT-6 into the culture medium . There was no evidence for increased virulence of the two rBCG strains when we made a comparison between them and BCG with regard to organ bacterial loads, lung histology, and survival time . rBCG-1 induced significantly higher specific antibody titers and stronger cellular immune response than BCG, whereas rBCG-2 had immunogenicity similar to that of the parental BCG strain . Both rBCG-1 and rBCG-2 conferred marked protection against M . tuberculosis infection, yet in terms of protective efficacy, they showed no significant improvements upon conventional BCG vaccine.

J Microbiol Methods, 2003 May, 53(2), 221 - 33
New biochip technology for label-free detection of pathogens and their toxins; Grow AE et al.; microSERS is a new biochip technology that uses surface-enhanced Raman scattering (SERS) microscopy for label-free transduction . The biochip itself comprises pixels of capture biomolecules immobilized on a SERS-active metal surface . Once the biochip has been exposed to the sample and the capture biomolecules have selectively bound their ligands, a Raman microscope is used to collect SERS fingerprints from the pixels on the chip . SERS, like other whole-organism fingerprinting techniques, is very specific . Our initial studies have shown that the Gram-positive Listeria and Gram-negative Legionella bacteria, Bacillus spores and Cryptosporidium oocysts can often be identified at the subspecies/strain level on the basis of SERS fingerprints collected from single organisms . Therefore, pathogens can be individually identified by microSERS, even when organisms that cross-react with the capture biomolecules are present in a sample . Moreover, the SERS fingerprint reflects the physiological state of a bacterial cell, e.g., when pathogenic Listeria and Legionella were cultured under conditions known to affect virulence, their SERS fingerprints changed significantly . Similarly, nonviable (e.g., heat- or UV-killed) microorganisms could be differentiated from their viable counterparts by SERS fingerprinting . Finally, microSERS is also capable of the sensitive and highly specific detection of toxins . Toxins that comprised as little as 0.02% by weight of the biomolecule-toxin complex produced strong, unique fingerprints when spectra collected from the complexes were subtracted from the spectra of the uncomplexed biomolecules . For example, aflatoxins B(1) and G(1) could be detected and individually identified when biochips bearing pixels of antibody or enzyme capture biomolecules were incubated in samples containing one or both aflatoxins, and the spectra were then collected for 20 s from an area of the biomolecule pixel approximately 1 microm in diameter . In the future, we plan to investigate the use of hyperspectral imaging Raman microscopy for collecting fingerprints from all the pixels on the biochip, individually yet simultaneously, to enable the rapid detection of diverse pathogens and their toxins in a sample, using a single biochip.

Anal Biochem, 2003 Mar 15, 314(2), 227 - 34
Isothermal titration calorimetric procedure to determine protein-metal ion binding parameters in the presence of excess metal ion or chelator; Nielsen AD et al.; Determination of binding parameters for metal ion binding to proteins usually requires preceding steps to remove protein-bound metal ions . Removal of bound metal ions from protein is often associated with decreased stability and inactivation . We present two simple isothermal titration calorimetric procedures that eliminate separate metal ion removal steps and directly monitor the exchange of metal ions between buffer, protein, and chelator . The concept is to add either excess chelator or metal ion to the protein under investigation and subsequently titrate with metal ion or chelator, respectively . It is thereby possible in the same experimental trial to obtain both chelator-metal ion and protein-metal ion binding parameters due to the different thermodynamic "fingerprints" of chelator and protein . The binding models and regression routines necessary to analyze the corresponding binding isotherms have been constructed . Verifications of the models have been done by titrations of mixtures of calcium chelators (BAPTA, HEDTA, and EGTA) and calcium ions and they were both able to account satisfactorily for the observed binding isotherms . Therefore, it was possible to determine stoichiometric and thermodynamic binding parameters . In addition, the concept has been tested on a recombinant alpha-amylase from Bacillus halmapalus where it proved to be a consistent procedure to obtain calcium binding parameters.

Int J Dermatol, 2003 Mar, 42(3), 197 - 200
Incidence of cutaneous tuberculosis in patients with organ tuberculosis; Kivanc-Altunay I et al.; BACKGROUND: Tuberculosis continues to be a health problem in some countries . The development of resistance to antituberculitic drugs and the increase in diseases and conditions associated with immunodeficiency such as AIDS and chemotherapy have caused tuberculosis to increase recently . As a result, the incidence of cutaneous tuberculosis has been increasing as well . AIM: To detect cutaneous tuberculosis in patients with organ tuberculosis and to establish some characteristics of the relation between organ and cutaneous TB . MATERIAL AND METHODS: A total of 370 patients (145 females and 225 males), aged 2-76 years (mean age 27.5), enrolled for this screening study . These patients were hospitalized patients who already had pulmonary or extrapulmonary tuberculosis diagnosed before admission . All patients underwent a general skin examination, and, if needed, cutaneous biopsies were taken from involved skin areas . RESULTS: Three hundred and forty-seven (93.78%) out of 370 patients had pulmonary tuberculosis only or in association with one of other organ tuberculoses . Twenty-three patients had extrapulmonary TB: nine were TB adenitis, six were TB peritonitis, three were bone tuberculosis, and five were TB meningitides . Of 370 patients, only 13 (3.51%) had cutaneous TB: seven scrofuloderma (SCD; 2.16%), four lupus vulgaris (LV; 1.35%), one LV and SCD, and one Bacille Calmette-Guerin (BCG) adenitis (0.027%) . Cutaneous tuberculosis was observed in seven out of 260 patients with parenchymal tuberculosis (2.96%) . Four out of nine patients with TB adenitis (44.4%), one out of 12 pulmopleuretic (8.3%), and one out of 67 pleuresic patients (1.40%) had cutaneous TB as well . Mean age of the 13 patients was 32.46 years: mean age of SCD and LV was 24.8 and 48 years, respectively . The one patient with BCG adenitis was 7 months old . Five (62.5%) out of eight patients with SCD, and only one (20%) out of five patients with LV were new cases . Four patients with SCD had a positive family history, while LV patients did not . CONCLUSIONS: Organ tuberculosis is rarely associated with cutaneous tuberculosis . Scrofuloderma and LV are the most frequent forms of skin TB associated with organ TB in this population . Tuberculosis adenitis is the organ TB that causes cutaneous TB most often among other organ tuberculoses . More than one form of cutaneous TB affected only one patient with pulmonary TB; therefore, it is very rare . Tuberculids were not observed in any of the patients.

Biochemistry, 2003 Apr 1, 42(12), 3457 - 63
A high-resolution NMR study of long-lived water molecules in both oxidation states of a minimal cytochrome c; Bertini I et al.; The interaction of water with oxidized and reduced cytochrome c from the Gram-positive bacterium Bacillus pasteurii (a 71-amino acid long monoheme cytochrome) is investigated through CLEANEX experiments and (15)N-edited ePHOGSY and Tr-ROESY experiments . It appears that a water molecule gives rise to dipolar cross-relaxation with the amide protons of Gly74 and Ile75, with a residence time longer than 0.4 ns, to account for a negative NOE . Such water molecule is present in both the oxidized and reduced species and in the X-ray structure . It appears to have a structural role . Other possible roles are discussed by comparison with the water molecules present in other c-type cytochromes . The amide proton of Cys35 is found to exchange rapidly with the solvent in the oxidized but not in the reduced protein, at variance with H/D exchange experiments, which probe a different time scale . The present data confirm that electron-transfer proteins evolved to minimize reorganization energy upon change of the oxidation state, even though the consequent variation of charge of the metal ion may induce some changes in the structure and/or dynamics of the protein.

Biotechnol Bioeng, 2003 Jun 5, 82(5), 517 - 24
Accelerated process development for protease production in continuous multi-stage cultures; Raninger A et al.; A fermentation process was developed and optimized for the production of a specific protease from Bacillus licheniformis PWD-1 . Media formulations were constructed and crucial environmental parameters were optimized to enhance growth and product formation . Process dynamics of substrate consumption, biomass-, product-, as well as by-product formation were determined under controlled conditions in a bioreactor . Using kinetic data from batch- and continuous-culture experiments, a fed-batch process was developed producing proteolytic activities 10 times those found during regular batch culture . In one stage continuous stirred tank culture protease formation was completely decoupled from sporulation . Shift experiments in one-stage continuous cultures led to the development of a two-stage continuous stirred tank fermentation process using optimized conditions for growth in the first stage and protease formation in the second stage . Accordingly, the basis for a continuous production of the enzyme on a pilot scale was accomplished .

Ecotoxicol Environ Saf, 2003 Mar, 54(3), 290 - 5
Sediment toxicity tests using benthic marine microalgae Cylindrotheca closterium (Ehremberg) Lewin and Reimann (Bacillariophyceae); Moreno-Garrido I et al.; A new method for sediment toxicity testing using marine benthic pennate noncolonial diatom (Cylindrotheca closterium, formerly Nitzschia closterium) has been developed . This microalgae showed a good growth rate during the experimental period, even when low enriched media were used . Sediment spiked with heavy metals {cadmium (Cd), copper (Cu), and lead (Pb)} was employed to determine the EC(50) values, using microalgal growth inhibition as the endpoint . The obtained results were as follows: Three heavy metals (Cd, Cu, and Pb), previously spiked on experimental sediment, were separately assayed in toxicity tests . The EC(50) values for these heavy metals in microalgal growth inhibition tests resulted to be 79 mg kg(-1) for Cd, 26 mg kg(-1) for Cu, and 29 mg kg(-1) for Pb (in experimental sediment) . The influence of sediment granulometry on the growth of microalgal population was also studied, finding that the growth of the microalgal population on media containing sediment with a relation sand-size:silt size of 9:1 was not different from optimal growth (occurring in media containing 100% sand-sized sediment) . The diatom C . closterium proved to be a suitable organism for sediment toxicity tests, due to its sensitivity and fast growth even in poorly enriched media.

Protein Expr Purif, 2003 Mar, 28(1), 190 - 5
Macroaffinity ligand-facilitated three-phase partitioning for purification of glucoamylase and pullulanase using alginate; Mondal K et al.; Starch-degrading enzymes glucoamylase (from Aspergillus niger), and pullulanase (from Bacillus acidopullulyticus) were purified using alginates (polysaccharides consisting of mannuronic acids and guluronic acids) by a recently developed technique called macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) . In this process, a crude preparation of the enzyme was mixed with alginate . On addition of appropriate amounts of ammonium sulfate and t-butanol, the alginate bound enzyme appeared as an interfacial precipitate between the lower aqueous and the upper t-butanol phase . Enzyme activity from this interfacial precipitate was recovered using 1M maltose . Glucoamylase and pullulanase were purified 20- and 38-fold with 83% and 89% activity recovery, respectively . Both the purified preparations showed a single band on SDS-PAGE.

Protein Expr Purif, 2003 Mar, 28(1), 63 - 8
Secretory expression in Escherichia coli and single-step purification of a heat-stable alkaline protease; Fu Z et al.; Bacteriocin release proteins (BRPs) can be used for the release of heterologous proteins from the Escherichia coli cytoplasm into the culture medium . The gene for a highly thermostable alkaline protease was cloned from Bacillus stearothermophilus F1 by the polymerase chain reaction . The recombinant F1 protease was efficiently excreted into the culture medium using E . coli XL1-Blue harboring two vectors: pTrcHis bearing the protease gene and pJL3 containing the BRPs . Both vectors contain the E . coli lac promoter-operator system . In the presence of 40 microM IPTG, the recombinant F1 protease and the BRP were expressed and mature F1 protease was released into the culture medium . This opens the way for the large-scale production of this protease in E . coli . The recombinant enzyme was purified through a one-step heat treatment at 70 degrees C for 3h and this method purified the protease to near homogeneity . The purified enzyme showed a pH optimum of 9.0, temperature optimum of 80 degrees C, and was stable at 70 degrees C for 24h in the pH range from 8.0 to 10.0 . The enzyme exhibited a high degree of thermostability with a half-life of 4 h at 85 degrees C, 25 min at 90 degrees C, and was inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF).

Protein Sci, 2003 Apr, 12(4), 861 - 70
Structure and dynamics of the DNA-binding protein HU of B . stearothermophilus investigated by Raman and ultraviolet-resonance Raman spectroscopy; Serban D et al.; The histone-like protein HU of Bacillus stearothermophilus (HUBst) is a 90-residue homodimer that binds nonspecifically to B DNA . Although the structure of the HUBst:DNA complex is not known, the proposed DNA-binding surface consists of extended arms that project from an alpha-helical platform . Here, we report Raman and ultraviolet-resonance Raman (UVRR) spectra diagnostic of subunit secondary structures and indicative of key side-chains lining the proposed DNA-binding surface . Raman conformation markers show that the DNA-binding arms of the dimer contain beta-stranded structure in excess (eight +/- two residues per subunit) of that reported previously . Important among side-chain markers are Met (701 cm(-1)), Ala (908 cm(-1)), Arg (1082 cm(-1)), and Pro (1457 cm(-1)) . The Ala marker undergoes a substantial shift (908 --> 893 cm(-1)) on deuteration of alanyl peptide sites, indicating a coupled side-chain/main-chain mode of diagnostic value in the identification of exchange-protected alanines . A large subset of alanines (67%) in the alpha-helical core exhibits robust resistance to exchange . A quantitative study of NH --> ND exchange exploiting newly identified amide II' markers of helical (1440 cm(-1)) and nonhelical (1472 cm(-1)) conformations of HUBst indicates unexpected flexibility at the dimer interface, which is manifested in rapid exchange of 80% of peptide sites . The results establish a basis for subsequent Raman and UVRR investigations of HUBst:DNA complexes and provide a framework for applications to other DNA-binding architectural proteins.

Proc Natl Acad Sci U S A, 2003 Apr 1, 100(7), 3895 - 900 Epub 2003 Mar 20.
Processive DNA synthesis observed in a polymerase crystal suggests a mechanism for the prevention of frameshift mutations; Johnson SJ et al.; DNA polymerases replicate DNA by adding nucleotides to a growing primer strand while avoiding frameshift and point mutations . Here we present a series of up to six successive replication events that were obtained by extension of a primed template directly in a crystal of the thermostable Bacillus DNA polymerase I . The 6-bp extension involves a 20-A translocation of the DNA duplex, representing the largest molecular movement observed in a protein crystal . In addition, we obtained the structure of a "closed" conformation of the enzyme with a bound triphosphate juxtaposed to a template and a dideoxy-terminated primer by constructing a point mutant that destroys a crystal lattice contact stabilizing the wild-type polymerase in an "open" conformation . Together, these observations allow many of the steps involved in DNA replication to be observed in the same enzyme at near atomic detail . The successive replication events observed directly by catalysis in the crystal confirm the general reaction sequence deduced from observations obtained by using several other polymerases and further refine critical aspects of the known reaction mechanism, and also allow us to propose new features that concern the regulated transfer of the template strand between a preinsertion site and an insertion site . We propose that such regulated transfer is an important element in the prevention of frameshift mutations in high-fidelity DNA polymerases . The ability to observe processive, high-fidelity replication directly in a crystal establishes this polymerase as a powerful model system for mechanistic studies in which the structural consequences of mismatches and DNA adducts are observed.

Cancer Res, 2003 Mar 15, 63(6), 1280 - 7
Development and preclinical evaluation of a Bacillus Calmette-Guérin-MUC1-based novel breast cancer vaccine; Chung MA et al.; Due to the high incidence of breast cancer and associated mortality rate,the development of an effective vaccine may be beneficial for the prevention or adjuvant treatment of this malignancy . We have constructed a novel breast cancer vaccine, Bacillus Calmette-Guerin (BCG)-hIL2MUC1, that consists of BCG and expresses a truncated form of MUC1 and human interleukin (IL)-2 . In vitro analysis of the BCG-hIL2MUC1 construct confirmed coexpression of MUC1 and human IL-2 . The ability of BCG-hIL2MUC1 to inhibit breast cancer growth was evaluated in hu-PBL-SCID mice (severe combined immunodeficient mice reconstituted with 50 x 10(6) human peripheral blood lymphocytes) that received three biweekly injections of BCG-hIL2MUC1 (0.5 colony-forming unit) . Control animals received PBS, MUC1 peptide (100 microg), or empty vector BCG-261 (0.5 colony-forming unit) vaccination . After immunization, hu-PBL-SCID mice (n = 8 in each group) were xenografted with 4 x 10(6) ZR75-1 human breast cancer cells . Whereas mice receiving the control vaccines developed a tumor, only 87% of BCG-hIL2MUC1-immunized animals developed a palpable tumor with a slower rate of tumor growth (P < 0.001) . Histological analysis of the primary tumors in BCG-hIL2MUC1-immunized animals revealed areas of reduced MUC1 expression . CD8-positive human lymphocytes were detected only in tumors grown in BCG-hIL2MUC1-immunized animals . These results imply a critical role of coexpressed IL-2 and MUC1 in eliciting tumor-specific immune response . To our knowledge, this is the first report of BCG engineered to express a tumor-associated antigen . Our results suggest that BCG-hIL2MUC1 immunization inhibited breast cancer growth in hu-PBL-SCID mice . Therefore, BCG-hIL2MUC1 may be a promising candidate as a breast cancer vaccine.

J Immunol, 2003 Apr 1, 170(7), 3696 - 706
Conservation of nonpeptide antigen recognition by rhesus monkey V gamma 2V delta 2 T cells; Wang H et al.; We have previously found that monkey Vgamma2Vdelta2(+) T cells mount adaptive immune responses in response to Mycobacterium bovis bacillus Calmette-Guerin infections . We have now analyzed rhesus monkey gammadelta T cell responses to nonpeptide Ags and superantigens . Like human Vgamma2Vdelta2(+) T cells, rhesus monkey gammadelta T cells are stimulated when exposed to prenyl pyrophosphate, bisphosphonate, and alkylamine Ags . Responsiveness was limited to gammadelta T cells expressing Vgamma2Vdelta2 TCRs . Rhesus monkey Vgamma2Vdelta2(+) T cells also responded to the superantigen, staphyloccocal enterotoxin A . Sequencing of the rhesus monkey Vgamma2Vdelta2 TCR revealed a strong sequence homology to human Vgamma2Vdelta2 TCR that preserves important sequence motifs . Moreover, chimeric TCRs that pair human Vgamma2 with monkey Vdelta2 and monkey Vgamma2 with human Vdelta2 retain reactivity to nonpeptide Ags and B cell lymphomas . A molecular model of the rhesus monkey Vgamma2Vdelta2 TCR has a basic region in the complementarity-determining region 3 binding groove that is similar to that seen in the human Vgamma2Vdelta2 TCR and preserves the topology of the complementarity-determining region loops . Thus, recognition of nonpeptide prenyl pyrophosphate, bisphosphonate, and alkylamine Ags is conserved in primates suggesting that primates can provide an animal model for human gammadelta T cell Ag responses.

Biochem Biophys Res Commun, 2003 Mar 28, 303(1), 31 - 6
Cloning of a novel one-repeat calcium channel-like gene; Arias JM et al.; We describe the cloning of a cDNA from a human testis library that encodes a novel protein with similarity to one repeat of voltage-gated Ca(2+) channels (Ca(v)) . Northern and dot blot analyses indicate that the novel Ca(v)-like gene is expressed predominantly in testis and at lower levels in many other tissues . Heterologous expression of the Ca(v)-like protein did not lead to the induction of any detectable ionic current and failed to modify intracellular Ca(2+) concentrations . Similar one-repeat Ca(v)-like proteins have been cloned from Bacillus, Mus, and Homo, and appear to encode ion channels involved in renal function, axis determination, and sperm motility.

Carbohydr Res, 2003 Mar 28, 338(7), 687 - 92
Carbohydrate esterase family 4 enzymes: substrate specificity; Caufrier F et al.; The substrate specificity of selected enzymes classified under Carbohydrate Esterase family 4 (CE4) has been examined . Chitin deacetylase from Mucor rouxii and both a native and a truncated form of acetyl xylan esterase from Streptomyces lividans were found to be active on both xylan and several soluble chitinous substrates . Furthermore, the activities of all enzymes examined were significantly increased in the presence of Co(2+) when chitinous substrates were employed . However, the presence of this metal ion did not result in enhancing the activities of the enzymes when xylan was used as substrate . An acetyl xylan esterase from Bacillus pumilus, classified under Carbohydrate Esterase family 7, was found to be inactive towards all chitinous substrates tested . Finally, all enzymes examined were inactive towards cell wall peptidoglycan.

FEMS Microbiol Lett, 2003 Mar 14, 220(1), 127 - 31
Isolation and characterisation of a novel bacteriocin produced by Bacillus thuringiensis strain B439; Ahern M et al.; Bacillus thuringiensis strain B439 produces a bacteriocin-like inhibitory substance in its growth medium . This antimicrobial peptide, referred to as thuricin 439, acts as a bacteriocidal peptide and exhibits an apparent narrow range of inhibitory activity, essentially only affecting growth of Bacillus cereus and B . thuringiensis strains . It remains active over a relatively wide pH and temperature range, showing no loss of activity following heat treatments up to 80 degrees C . Purification of thuricin 439 was achieved using several chromatographic steps, which resulted in the identification of two peptides with inhibitory activity . These two peptides were shown to possess identical N-terminal sequences, but different molecular masses.

Bioconjug Chem, 2003 Mar-Apr, 14(2), 440 - 8
Generation of a functional monomolecular protein lattice consisting of an s-layer fusion protein comprising the variable domain of a camel heavy chain antibody; Pleschberger M et al.; Crystalline bacterial cell surface layer (S-layer) proteins are composed of a single protein or glycoprotein species . Isolated S-layer subunits frequently recrystallize into monomolecular protein lattices on various types of solid supports . For generating a functional protein lattice, a chimeric protein was constructed, which comprised the secondary cell wall polymer-binding region and the self-assembly domain of the S-layer protein SbpA from Bacillus sphaericus CCM 2177, and a single variable region of a heavy chain camel antibody (cAb-Lys3) recognizing lysozyme as antigen . For construction of the S-layer fusion protein, the 3'-end of the sequence encoding the C-terminally truncated form rSbpA(31)(-)(1068) was fused via a short linker to the 5'-end of the sequence encoding cAb-Lys3 . The functionality of the fused cAb-Lys3 in the S-layer fusion protein was proved by surface plasmon resonance measurements . Dot blot assays revealed that the accessibility of the fused functional sequence for the antigen was independent of the use of soluble or assembled S-layer fusion protein . Recrystallization of the S-layer fusion protein into the square lattice structure was observed on peptidoglycan-containing sacculi of B . sphaericus CCM 2177, on polystyrene or on gold chips precoated with thiolated secondary cell wall polymer, which is the natural anchoring molecule for the S-layer protein in the bacterial cell wall . Thereby, the fused cAb-Lys3 remained located on the outer S-layer surface and accessible for lysozyme binding . Together with solid supports precoated with secondary cell wall polymers, S-layer fusion proteins comprising rSbpA(31)(-)(1068) and cAbs directed against various antigens shall be exploited for building up monomolecular functional protein lattices as required for applications in nanobiotechnology.

PDA J Pharm Sci Technol, 2003 Jan-Feb, 57(1), 3 - 11
Effect of carrier materials on the resistance of spores of Bacillus stearothermophilus to gaseous hydrogen peroxide; Sigwarth V et al.; The testing of the H2O2 decontamination process using spores of Bacillus stearothermophilus has gained widespread acceptance . Usually, commercially available Biological Indicators (BIs) with a specified resistance to H2O2 are challenged to qualify the process . The question arises whether the resistance of test spores is dependant on the type of carrier material and whether the resistance is representative for the system under test . The objective of the study is to quantify the effect of different carrier materials on the resistance of spores of Bacillus stearothermophilus to H2O2 . Materials from which isolators were built, as well as those used in disposables during daily work were investigated . These materials were inoculated with 106 spores of Bacillus stearothermophilus (ATCC 7953) . The spore resistance was tested to a well defined H2O2 decontamination cycle by determining the D-value using the "Fractional Negative" method . This paper reports on the effect of different carrier materials to the resistance of the test organism against H2O2 . Various materials have significantly increased resistance of the spores and should be avoided in isolator systems . If commercially available BIs are used for process qualification, the resistance of the BI used, the fluctuation in resistance caused by isolator materials, the required log reduction, and at least the bioload of isolator surfaces need to be known.

Lett Appl Microbiol, 2003, 36(4), 251 - 7
Importance of Gram-positive naphthalene-degrading bacteria in oil-contaminated tropical marine sediments; Zhuang WQ et al.; AIMS: The aim of this study was to isolate, characterize and evaluate the importance of naphthalene-degrading bacterial strains from oil-contaminated tropical marine sediments . METHODS AND RESULTS: Three Gram-positive naphthalene-degrading bacteria were isolated from oil-contaminated tropical intertidal marine sediments by direct isolation or enrichment using naphthalene as the sole source of carbon and energy . Bacillus naphthovorans strain MN-003 can also grow on benzene, toluene, xylene and diesel fuel while Micrococcus sp . str . MN-006 can also grow on benzene . Staphylococcus sp . str . MN-005 can only degrade naphthalene and was not able to use the other aromatic hydrocarbons tested . Strain MN-003 possessed the highest maximal specific growth rate with naphthalene as sole carbon source . An enrichment culture fed with naphthalene as sole carbon source exhibited a significant increase in the relative abundances of the three isolates after 21 days of incubation . The three isolates constituted greater than 69% of the culturable naphthalene-degrading microbial community . Strain MN-003 outcompeted and dominated the other two isolates in competition studies involving batch cultures inoculated with equal cell densities of the three isolates and incubated with between 1 and 10 mg l-1 of naphthalene . CONCLUSIONS: Three Gram-positive naphthalene-degrading bacteria were successfully isolated from oil-contaminated tropical marine sediments . Gram-positive bacteria might play an important role in naphthalene degradation in the highly variable environment of oil-contaminated tropical intertidal marine sediments . Among the three isolates, strain MN-003 has the highest maximal specific growth rate when grown on naphthalene, and outgrew the other two isolates in competition experiments . SIGNIFICANCE AND IMPACT OF THE STUDY: This research will aid in the development of bioremediation schemes for oil-contaminated marine environments . Strain MN-003 could potentially be exploited in such schemes.

Bull World Health Organ, 2003, 81(1), 61 - 70 Epub 2003 Mar 11.
Immunization of children at risk of infection with human immunodeficiency virus; Moss WJ et al.; This paper reviews the English language literature on the safety, immunogenicity and effectiveness in children infected with the human immunodeficiency virus (HIV) of vaccines currently recommended by WHO for use in national immunization programmes . Immunization is generally safe and beneficial for children infected with HIV, although HIV-induced immune suppression reduces the benefit compared with that obtained in HIV-uninfected children . However, serious complications can occur following immunization of severely immunocompromised children with bacillus Calmette-Gu rin (BCG) vaccine . The risk of serious complications attributable to yellow fever vaccine in HIV-infected persons has not been determined . WHO guidelines for immunizing children with HIV infection and infants born to HIV-infected women differ only slightly from the general guidelines . BCG and yellow fever vaccines should be withheld from symptomatic HIV-infected children . Only one serious complication (fatal pneumonia) has been attributed to measles vaccine administered to a severely immunocompromised adult . Although two HIV-infected infants have developed vaccine-associated paralytic poliomyelitis, several million infected children have been vaccinated and the evidence does not suggest that there is an increased risk . The benefits of measles and poliovirus vaccines far outweigh the potential risks in HIV-infected children . The policy of administering routine vaccines to all children, regardless of possible HIV exposure, has been very effective in obtaining high immunization coverage and control of preventable diseases . Any changes in this policy would have to be carefully examined for a potential negative impact on disease control programmes in many countries.

J Ethnopharmacol, 2003 Apr, 85(2-3), 283 - 7
Inhibitory effect of aqueous extract from the gall of Rhus chinensis on alpha-glucosidase activity and postprandial blood glucose; Shim YJ et al.; The present study examined the inhibitory effect of aqueous extract from the gall of Rhus chinensis (AEGRC) on alpha-glucosidase activity, an enzyme responsible for digestion of carbohydrate to monosaccharides in the process of intestinal absorption . AEGRC inhibited Bacillus alpha-glucosidase acitvity with an IC(50) of 0.9 micro g/ml . Its inhibition on alpha-glucosidase was determined to be noncompetitive and reversible when the enzyme-substrate mixture was simultaneously treated with AEGRC as an inhibitor . In addition, when it was orally administered to rats with sucrose (2g/kg), AEGRC (250-1000mg/kg) significantly suppressed the increase of blood glucose levels after sucrose loading in a dose dependent manner . These results suggest that AEGRC might exert anti-diabetic effect by suppressing carbohydrate absorption from intestine, and thereby reducing the postprandial increase of blood glucose.

J Food Prot, 2003 Mar, 66(3), 441 - 5
Evaluation and interlaboratory validation of a selective agar for phosphatidylinositol-specific phospholipase C activity using a chromogenic substrate to detect Listeria monocytogenes from foods; Jinneman KC et al.; Phosphatidylinositol-specific phospholipase C (PI-PLC) activity is a potential virulence factor and is exhibited only by the Listeria species Listeria monocytogenes and Listeria ivanovii . A chromogenic substrate for the direct detection of PI-PLC activity is available in a new medium (BCM L . monocytogenes plating agar) . The use of a chromogenic substrate offers a mechanism with which to directly screen for L . monocytogenes and L . ivanovii other than the esculin used in Oxford (OXF) and Palcam (PAL) agars, which screen for all Listeria species . The specificity levels of BCM plating agar and of BCM confirmation and rhamnose agars were evaluated with 107 Listeria and 10 Bacillus species isolates . In addition, BCM L . monocytogenes plating agar was compared with standard Listeria selective agars (OXF and PAL agars) with regard to the recovery of L . monocytogenes from 2,000 food and environmental samples obtained from eight participating laboratories . A Listeria species was isolated from at least one of the agars in 209 analyses, and L . monocytogenes was isolated in 135 of these analyses . In 27 of the analyses in which L . monocytogenes was isolated, one or more of the selective differential agars used failed to isolate L . monocytogenes, and therefore the results of these analyses were discrepant . Relative to a reference method involving the use of all three agars (OXF, PAL, and BCM agars), the OXF-BCM, PAL-BCM, and OXF-PAL combinations had sensitivities of 99.3, 99.2, and 90.2%, respectively . In statistical analyses of the different combinations of agars, the OXF-BCM and BCM-PAL combinations were found to be superior to the OXF-PAL combination for the detection of L . monocytogenes.

Int J Med Microbiol, 2003 Feb, 292(7-8), 441 - 51
Novel vaccination strategies based on recombinant Mycobacterium bovis BCG; Dietrich G et al.; In this manuscript, we will review the utilization of Mycobacterium bovis Bacille Calmette-Guerin (BCG) as a vaccine against tuberculosis (TB) and as a carrier system for heterologous antigens . BCG is one of the most widely used vaccines . Novel techniques in genome manipulation allow the construction of virulence-attenuated recombinant (r)-BCG strains that can be employed as homologous vaccines, or as heterologous antigen delivery systems, for priming pathogen-specific immunity against infectious diseases, including TB . Several approaches are available for heterologous antigen expression and compartmentalization in BCG and recent findings show the potential to modulate and direct the immune responses induced by r-BCG strains as desired . Recent achievements in complete genome analysis of various target pathogens, combined with a better understanding of protective pathogen-specific immune responses, form the basis for the rational design of a new generation of recombinant mycobacterial vaccines against a multitude of infectious diseases.

Heredity, 2003 Feb, 90(2), 141 - 9
Host-plant-associated genetic differentiation in Northern French populations of the European corn borer; Martel C et al.; The phytophagous insects that damage crops are often polyphagous, feeding on several types of crop and on weeds . The refuges constituted by noncrop host plants may be useful in managing the evolution in pest species of resistance to the Bacillus thuringiensis toxins produced by transgenic crops . However, the benefits of these refuges may be limited because host-plant diversity may drive genetic divergence and possibly even host-plant-mediated sympatric speciation . The European corn borer, Ostrinia nubilalis Hubner (Lepidoptera: Crambidae), is the main pest of maize in Europe and North America, where it was introduced early in the 20th century . It has a wide host range but feeds principally on mugwort (Artemisia vulgaris L.) and maize (Zea mays L.) . O . nubilalis is found on mugwort only in the northern part of France, whereas it is found on maize throughout France . The extent of genetic variation at allozyme markers was investigated in populations collected from the two host plants over the entire geographical distribution of the European corn borer on mugwort in France . Allelic differentiation between pairs of populations and hierarchical analyses of pools of samples from each host plant indicate that the group of populations feeding on maize differed from the group of populations feeding on mugwort . Our results suggest (1) host-plant-related divergent selection at the genomic region surrounding the Mpi locus and (2) limited gene flow between the populations feeding on mugwort and those infesting maize fields . These data indicate that adults emerging from mugwort would not be useful for managing the evolution of resistance to the B . thuringiensis toxins in European corn borer populations.

Microbiology, 2003 Mar, 149(Pt 3), 729 - 37
Correlation of the rate of protein synthesis and the third power of the RNA : protein ratio in Escherichia coli and Mycobacterium tuberculosis; Cox RA; In order to further understand the different physiological states of the tubercle bacillus, a frame of reference was sought by first correlating the macromolecular compositions of Escherichia coli with specific growth rates and also with the rates of protein synthesis . Data for DNA : protein : RNA were converted to the average amounts of DNA {m(DNA(av))}, protein {m(p(av))} and RNA {m(RNA(av))} per cell . The specific growth rate micro was found to be directly proportional to m(RNA(av))/m(p(av)) . The specific protein synthesis rate per average cell {omega(p(av))} was shown to be directly proportional to the third power of the ratio m(RNA(av))/m(p(av)) which reflects the ribosome concentration . The equations derived were shown apply to both E . coli ( micro =1.73 h(-1)) and Mycobacterium bovis BCG ( micro =0.029 h(-1)).

FEBS Lett, 2003 Mar 13, 538(1-3), 29 - 34
A cadherin-like protein functions as a receptor for Bacillus thuringiensis Cry1Aa and Cry1Ac toxins on midgut epithelial cells of Bombyx mori larvae; Hara H et al.; Aminopeptidase N (APN) and cadherin-like protein (BtR175) from Bombyx mori larvae were examined for their roles in Cry1Aa- and Cry1Ac-induced lysis of B . mori midgut epithelial cells (MECs) . APNs and BtR175 were present in all areas of the midgut, were particularly abundant in the posterior region, and were found only on columnar cell microvilli and not on the lateral membrane that makes cell-cell contacts . This distribution was in accordance with the distribution of Cry1A-susceptible MECs in the midgut . The lytic activity of Cry1Aa and Cry1Ac on collagenase-dissociated MECs was linearly dependent on toxin concentration . Although pre-treatment of MECs with anti-BtR175 antibody was observed to partially inhibit the lytic activity exerted by 0.1-1 nM Cry1Aa toxin or 5 nM Cry1Ac toxin, no significant inhibition was observed when MECs were pre-treated with anti-APN antibody . These results suggest that BtR175 functions as a major receptor for Cry1A toxins in the midgut of B . mori larvae.

Tunis Med, 2002 Oct, 80(10), 645 - 9
{Abdomino-pelvic actinomycosis: a case report}; Bedoui R et al.; The actinomycosis is a chronic suppurative granulomatosis disease . It is owed to a bacillus gram positive; actinomyces israelli . The cervical and thoracic localizations are most frequent . The digestive localization represents 20% of cases . It interest very rarely the pelvis and the genital tracts . We bring back the observation of a patient old of 30 years admitted for mass abdominal . To the exam, the patient had a sensibility of the left hypochondriac area and we found a mass of 6 cm of diameter . To the rectal touch, we found a mass in the bag of Douglas . The echography and the computed tomography revealed a collection under the spleen and a pelvic collection . A rectotomy is performed . The bacteriological study isolates actinomyces israelli . The collection under the spleen is drained under radiological control . Actinomyces israelli is also recovered in the pus brought back by the puncture . The patient is treated by Penicillin . The patient had a favourable evolution . No etiology is found at this patient . For this observation, the collection was accessible to a drainage permitting the diagnosis and the treatment of the actinomycosis while avoiding a mutilated surgery.

World J Gastroenterol, 2003 Mar, 9(3), 516 - 20
Virulence of water-induced coccoid Helicobacter pylori and its experimental infection in mice; She FF et al.; AIM: To explore the virulence and the infectivity of coccoid Helicobacter pylori (H . pylori) transformed from spiral form by exposure to sterile tap water . METHODS: Three strains of H . pylori, isolated from gastric biopsy specimens of confirmed peptic ulcer, were converted from spiral into coccoid form by exposure to sterile tap water . Both spiral and coccoid forms of H . pylori were tested for the urease activity, and the adherence to Hep-2 cells . The presence of flagella was examined under electron microscopy . In the experimental animal infection, the spiral and coccoid forms of H . pylori originated from the same strain F49 were inoculated intragastrically into BALB/c mice respectively four times at a 3-day interval . Half of the mice from each group were sacrificed at Day 21 and Day 28 after the last inoculation . Histology and H . pylori colonization were detected by urease test of gastric mucosa, cultures of H . pylori, and electron microscopy and so on . RESULTS: The urease activity and the ability of adherence to Hep-2 cells were found to be lower in coccoid H . pylori than that in its spiral form . For example, the transformation in strain F(44) led to a significant decrease of the adherence rate and adherence index from 70.0+/-5.3 % to 30.2+/-3.5 % (P<0.01), and from 2.6+/-0.4 to 0.86+/-0.3 (P<0.01), respectively . The flagella of coccoid H . pylori were observed under electron microscope . In the experimental infection in mice, the positive rate of gastric mucosa urease test was 93.8 % (15/16) in the group infected by spiral H . pylori and 50 % (8/16) in the group infected by coccoid H . pylori, and the estimated coccoid H . pylori colony number was 1.75 vs 0.56 . The positive rates of H . pylori culture were 87.5 % (14/16) in spiral H . pylori group and 68.8 % (11/16) in coccoid H . pylori group . There was no significant difference in either urease test or bacterial culture rate between the groups examined at Day 21 and Day 28 after inoculation . Electron microscopic examination of the samples taken from both groups showed the adherence of H . pylori in spiral, bacillary and coccoid shapes to the epithelial cells of gastric wall . Histological examination showed the occurrence of gastric mucosal injury as indicated by various degrees of erosion, ulcer, and inflammatory cell infiltration . Mucosal injury was slighter in the mice infected by coccoid H . pylori . No positive result was obtained in the control group that received intragastrical administration of sterile tap water . CONCLUSION: Although the virulence of coccoid H . pylori induced by water decrease, coccoid H . pylori still remains a considerable urease activity and the adhering ability to epithelial cells . Furthermore, the flagella, an important component responsible for bacterial movement and infection, were still observed as a cellular structure of coccoid H . pylori under electron microscope . The coccoid H . pylori induced by water is capable of colonizing in gastric mucosa and causing gastrititis in mice.

Biotechnol Appl Biochem, 2003 Apr, 37(Pt 2), 165 - 71
Regulatory effects of alanine-group amino acids on serine alkaline protease production by recombinant Bacillus licheniformis; Calik P et al.; Influences of the concentration and addition time of alanine-group amino acids, i.e . alanine, leucine and valine, on serine alkaline protease (SAP) synthesis were investigated by Bacillus licheniformis (DSM 1969) carrying pHV1431:: sub C in a defined medium to identify the amino acids creating intracellular reaction-rate limitation in SAP production . While the precursors of alanine-group amino acids, pyruvate and alanine, did not affect SAP production considerably within the range 0-15 mM, the addition of leucine decreased both SAP production and cell formation, because of the inhibition of valine synthesis . Although valine inhibits reactions starting with pyruvate towards 2-oxo-isovalerate, due to conversion of valine into 2-oxo-isovalerate and from 2-oxo-isovalerate to leucine, valine did not inhibit leucine synthesis . Val (7.5 mM) supply at t =0 h increased SAP activity to an activity of 1070 units.cm(-3) which was 1.3-fold higher than that of the reference production medium . The highest cell growth yield on substrate (Y (X/S)) was obtained as 0.24 g.g(-1) with the supply of alanine; and the highest product formation yield on substrate was obtained as 0.134 units.g(-1) with the supply of valine . By using the results obtained, strategies for increasing SAP production and complex medium design were also discussed.

Folia Microbiol (Praha), 2002, 47(6), 759 - 65
Differential effect of Bacillus firmus on immune response and enterocyte brush-border enzyme levels in BALB/c and B10.BR mice; Kozakova H et al.; A nonpathogenic bacterium of external environment possessing remarkable immunomodulatory activity, Bacillus firmus (BF) inactivated with formaldehyde, was given intragastrically to two genetically different mouse strains BALB/c (H-2d) and B10.BR/SnPh (B10.BR, H-2k) reared in conventional (CV) and B10.BR strain also in germ-free (GF) conditions . Repeated intragastric administration of BF (500 micrograms every other day over two weeks, starting at the age of 3 months) significantly enhanced intestinal IgA levels in CV BALB/c mice but did not affect intestinal IgA in CV B10.BR mice . In GF B10.BR mice, IgG levels in sera and intestinal washings increased after BF administration compared to CV B10.BR mice . In CV BALB/c mice, specific activity of enterocyte brush-border enzymes (lactase, gamma-glutamyltransferase, alkaline phosphatase) decreased after BF treatment; sucrase (sucrose alpha-glucosidase) activity was not affected . On the other hand, in B10.BR mice, specific activity of gamma-glutamyltransferase and dipeptidyl peptidase IV were higher after administration of BF in both CV and GF groups relative to untreated controls . The activities of lactase and glucoamylase (glucan 1,4-alpha-glucosidase) were significantly stimulated only in the group of GF B10.BR mice treated with formolized BF . The stimulation of immunoglobulin production after BF treatment was accompanied by changes in the levels of enterocyte brush-border enzymes; this responsiveness to BF treatment was genetically regulated.

Hepatogastroenterology, 2003 Jan-Feb, 50(49), 201 - 6
Analysis of the effect of IL-12 therapy on immunoregulatory T-cell subsets in patients with chronic hepatitis C infection; Barth H et al.; BACKGROUND/AIMS: Aim of this study was to investigate the influence of IL-12 therapy on the production of immunoregulatory type 1/type 2-cytokines by peripheral blood mononuclear cells from patients with chronic hepatitis C . METHODOLOGY: Peripheral blood mononuclear cells were isolated from 12 patients with chronic HCV infection before and after eight weeks of IL-12 application (0.03-0.5 micrograms/kg body weight) . Peripheral blood mononuclear cells from 40 healthy blood donors served as a control group . The peripheral blood mononuclear cells were incubated for seven days with antigens stimulating specifically type 1 (tuberculin purified protein derivative) or type 2 T-cells (tetanus-toxoid) . Furthermore, BCG (bacille Calmette-Guerin) was added to the cultures known to activate macrophages/antigen-presenting cells . Supernatants of peripheral blood mononuclear cells were analyzed for tuberculin purified protein derivative-induced production of the type 1 cytokine IFN-gamma, tetanus-toxoid-induced production of the type 2 cytokine IL-5 and the BCG-induced production of TNF-alpha by a double sandwich ELISA . RESULTS: IL-12 therapy hardly influenced the regulatory type 1/type 2 T-cell reactivity . In contrast, BCG induced secretion of TNF-alpha was significantly higher after eight weeks of IL-12 therapy (4348 +/- 3083 pg/mL) than before treatment (1559 +/- 988 pgmL, p < 0.01) . Clinically, serum alanine transaminase levels significantly decreased during IL-12 treatment but HCV-RNA persisted in all patients . CONCLUSIONS: IL-12 therapy in patients with HCV does not alter the production of regulatory cytokines produced by type 1/type 2 TH cells . The significantly enhanced production of BCG-induced TNF-alpha may, however, indicate an activation of antigen presenting cells or natural killer cells.

Rheumatology (Oxford), 2003 Mar, 42(3), 473 - 80
Whipple arthritis: diagnosis by molecular analysis of synovial fluid--current status of diagnosis and therapy; Lange U et al.; Whipple's disease (WD) is an uncommon polysystem infectious disease . In the present report, we describe a patient who presented with a chronic illness consistent with WD and an avascular necrosis of the right hip joint . WD and its proposed causative bacillus, Tropheryma whippelii, was identified by molecular analysis (polymerase chain reaction) in bacterial DNA extracted from the synovial fluid . The diagnosis was additionally confirmed by upper gastrointestinal endoscopy and a small bowel biopsy with macrophages positive for periodic acid-Schiff reagent demonstrated by light and electron microscopy . This demonstrates that WD can be diagnosed without tissue biopsy . False diagnosis of the polymorphous signs and symptoms of WD can lead to invalidism and even death, whereas correct therapy leads to a cure in most cases . Thus, the current status of diagnosis and therapy is of key importance in the treatment of WD.

Ann Trop Med Parasitol, 2002 Dec, 96 Suppl 2, S105 - 16
Control of the Aedes vectors of the dengue viruses and Wuchereria bancrofti: the French Polynesian experience; Lardeux F et al.; In most of the 130 islands of French Polynesia, the stenotopic mosquitoes Aedes aegypti (the main local vector for the viruses causing dengue) and Aedes polynesiensis (the main local vector of Wuchereria bancrofti) share many breeding sites in water containers such as discarded cans, coconut shells, buckets and water-storage pots and drums . In addition to selective application of insecticides, non-polluting methods of controlling these mosquitoes have been evaluated during the last decade in two main ecological situations: (1) villages, where Aedes breeding sites are typically peridomestic; and (2) flooded burrows of land crabs, the major source of Ae . polynesiensis throughout the South Pacific region . Large-scale trials of biological control agents, such as mosquito fish (Gambusia affinis and Poecilia reticulata) and copepods (Mesocyclops aspericornis), and of integrated-control strategies have demonstrated the efficacy of certain techniques and control agents against the target Aedes populations in some village situations . Generally, mechanical methods (the use of layers of polystyrene beads against mosquito larvae and pupae, and screening against adult mosquitoes) were more efficient than use of the biological control agents . By integrating several methods of control, mosquito densities (as measured by human-bait collections and larval surveys) were reduced significantly compared with the results of concurrent sampling from untreated villages, and control remained effective for months after the interventions ceased . In land-crab burrows, the first attempts to control Aedes larvae used bacterial agents (Bacillus thuringiensis) and predatory copepods gave disappointing results . Mesocyclops aspericornis could be an effective control agent if the burrows were constantly flooded, but most burrows dry out and refill periodically, so copepod populations do not survive . As it proved difficult to reach all corners of the long sinuous burrows with any control agent, larvicidal (chlorpyrifos-methyl) baits were developed for foraging crabs to carry into their burrows . This novel technique proved to be effective and could become the method of choice for treating crab burrows . Further research is underway to find the optimum (biological or chemical) larvicidal ingredient for the crab bait . Despite the ecological and logistical challenges of controlling the Aedes vectors of the dengue viruses and W . bancrofti in so many scattered islands, the French Polynesian experience indicates that relatively simple methods can be integrated and applied effectively and economically . Operationally, however, success also depends on a strong political commitment and on at-risk communities that are sufficiently motivated to maintain a good level of Aedes control.

Ann Trop Med Parasitol, 2002 Dec, 96 Suppl 2, S77 - 90
Cost-effectiveness of the use of vector control and mass drug administration, separately or in combination, against lymphatic filariasis; Krishnamoorthy K et al.; The costs and effects of two intervention strategies for the control of bancroftian filariasis-annual mass drug administrations (MDA) with a combination of diethylcarbamazine and ivermectin, with or without integrated vector control (VC)-were estimated in rural villages in South India . The aim was to compare the cost-effectiveness of MDA alone with that of MDA plus VC . Control of the local vector, Culex quinquefasciatus, was based on the application of polystyrene beads to cesspits, the treatment of drains with larvicidal Bacillus sphaericus and the stocking of wells with larvivorous fish . An itemized cost menu was used to cost MDA and MDA + VC, retrospectively . The annual transmission potential was used to assess the direct outcome of the disease-control methods, whereas the prevalence and intensity of microfilaraemia were used as indicators of the impact of each method . The per-capita costs were 1.49 U.S . dollars for two rounds of MDA, 1.70 U.S . dollars for 2 years of VC and, therefore, 3.19 U.S . dollars for 2 years of MDA + VC . Integration of VC with MDA did not appear to be cost-effective: it cost an estimated 1.80 U.S . dollars to stop an infective mosquito biting a villager using MDA alone but 3.32 U.S . dollars to achieve the same result using MDA + VC . Similarly, the cost to reduce the prevalence of microfilaraemia in a three-village group by 1% was only 96.62 U.S . dollars for MDA alone but 201.16 U.S . dollars when vector control was integrated . The implications of these results for the control and elimination of filariasis in Indian village communities, and the options for sharing and minimizing costs, are discussed.

Ying Yong Sheng Tai Xue Bao, 2002 Nov, 13(11), 1452 - 4
{Microbiological eco-regulation in Vc two-step fermentation}; Zhou B et al.; The growth and interaction of Gluconobacter axydans and Bacillus megaterium in Vc two step fermentation have been studied . The results showed that there is a kind of coordinate symbiosis between G . axydans and B . megaterium during the fermentation, which could promoted 2-Keto-L-gulonic acid (2KGA) synthesis . The efficiency on the synbiosis would vary as the conditions of fermentation were different and were improved by the environmental factors.

Mol Biol (Mosk), 2003 Jan-Feb, 37(1), 128 - 38
{DNA-{N4-cytosine}-methyltransferase from Bacillus amyloliquefaciens: mechanism of action derived from steady state kinetics}; Zinov'ev VV et al.; Kinetic analysis of methyl group transfer from S-adenosyl-L-methionine (SAM) to the 5'-GGATCC recognition site catalyzed by the DNA-{N4-cytosine}-methyltransferase from Bacillus amyloliquefaciens {EC 2.1.1.113} has shown that the dependence of the rate of methylation of the 20-meric substrate duplex on SAM and DNA concentration are normally hyperbolic, and the maximal rate is attained upon enzyme saturation with both substrates . No substrate inhibition is observed even at concentrations many times higher than the Km values (0.107 microM for DNA and 1.45 microM for SAM), which means that no nonreactive enzyme-substrate complexes are formed during the reaction . The overall pattern of product inhibition corresponds to an ordered steady-state mechanism following the sequence SAM decreases DNA decreases metDNA increases SAH increases (S-adenosyl-L-homocysteine) . However, more detailed numerical analysis of the aggregate experimental data admits an alternative order of substrate binding, DNA decreases SAM decreases, though this route is an order of magnitude slower.

J Clin Microbiol, 2003 Mar, 41(3), 1069 - 72
Detection and identification of Bartonella species pathogenic for humans by PCR amplification targeting the riboflavin synthase gene (ribC); Johnson G et al.; Several Bartonella species have now been implicated as human pathogens . The recovery of these fastidious organisms in the clinical microbiology laboratory remains difficult, and current methods are still relatively insensitive . Thus, the bartonellae are good candidates for detection by PCR . We have developed a PCR assay which uses a single primer pair targeting the riboflavin synthase gene (ribC) and detected six Bartonella species that have been implicated in human disease, B . henselae, B . quintana, B . bacilliformis, B . clarridgeiae, B . elizabethae, and B . vinsonii subsp . berkhoffii . Species identification is achieved simply by restriction enzyme digestion of the amplicon . This PCR assay appears to be specific for the Bartonella genus because it failed to amplify DNA from several other bacterial species.

J Invertebr Pathol, 2003 Feb, 82(2), 119 - 27
Distribution and diversity of cry genes in native strains of Bacillus thuringiensis obtained from different ecosystems from Colombia; Uribe D et al.; Colombia is a tropical country located at the north of South America . It is considered to be one of the most important countries in terms of its biodiversity worldwide . One hundred and eight soil samples obtained from agricultural crops and wild ecosystems were evaluated in terms of the presence of Bacillus thuringiensis (Bt) native strains . One hundred and eight different Bt strains were isolated and characterized by the presence of crystal proteins by SDS-PAGE and a multiplex PCR with general and specific primers for cry1 and cry3, cry7, and cry8 gene detection . Most of the Bt strains (73%) reacted with the cry1 general primers; 27.8% of the Bt strains reacted with cry3, cry7, and cry8 general primers and 17.8% of strains did not react with any of these two sets of primers . Thirty different PCR profiles were found in the strains with cry1 genes when they were analyzed with specific primers (cry1A to cry1F) . A high frequency of joint occurrence was observed for cry1Aa/cry1Ab, cry1Aa/cry1Ac, cry1Ab/cry1Ac, and cry1C/cry1D genes with a Pearson coefficient of 0.88, 0.74, 0.76, and 0.87, respectively . Other distinctive characteristics were found in the Colombian collection as the presence of 22.2% of native strains which presented, at the same time, lepidopteran and coleopteran active genes . Interesting relations were found as well between the cry gene distribution and the geographical areas sampled . Finally, some strains with moderate to high biopesticide activity against Spodoptera frugiperda (Lepidoptera) and Premnotrypes vorax (Coleoptera) insects were identified, this being important to explore future microbial strategies for the control of these crop pests in the region.

Tuberculosis (Edinb), 2002, 82(6), 261 - 6
Pattern recognition technique in immunological antigenic tests to identify Mycobacterium tuberculosis infection; Selvaraj R et al.; The importance of a diagnostic test that is simple and quick to identify Mycobacterium tuberculosis infection needs no emphasis . The tuberculin skin test (TST - 1 TU RT23) is the diagnostic tool for identifying M . tuberculosis infection at present . The test reaction on the skin is measured after 48-72 h . It is observed that often multi-modes are seen, when the reactions are drawn as a graph and the bimodality is seen very feebly . Because of the difficulties in the administration of TST, several serological tests were developed over three decades, but none of the studies showed the desired results . One study to evaluate purified protein derivative (PPD) antigen resulted in a claim of 80% sensitivity and 4% false-positivity rate (14), while other researchers were not able to obtain similar results . In addition, several problems were encountered due to the non-availability of antigens, and data analyses from an ELISA-based diagnostic test showed considerable overlap of distributions of optical density (OD) values among patients and healthy individuals (10) . Classical statistical techniques cannot explain the cause of these overlaps . Hence, an attempt is made in this article to resolve these difficulties by the pattern recognition technique (PRT) . The technique lies in splitting the data into clusters using a supervised algorithm . The data set is normally split into a training set, a test set and a validation set . The PRT gets "trained" through the training data set until the infected and uninfected groups of individuals are correctly classified . The training occurs based on an algorithm on the training set . On successful completion of the training, this technique is further tested and validated in the respective data sets . SETTING: A total of 273 finger-prick specimens were collected from five categories (Al, A2, B, C, D, E) of subjects not vaccinated with (bacille Calmette Guerin) (BCG) from Trivellore BCG Trial area adopted by the Tuberculosis Research Center, Chennai, India . OBJECTIVE: The study was conducted with the primary aim of evaluating purified antigens--r38 kDa, PPD and 30 kDa--for their usefulness as diagnostic tools and to test the applicability of the PRT in the evaluation of diagnostic tests . Individuals in two main categories (definitely not infected categories Al, A2 and D, and definitely infected categories B, E and C based on reaction to TST) were assembled for the purpose . RESULTS: The overall PRT performance of 30 kDa was 72.3% sensitivity and 90.9% specificity for identifying M . tuberculosis infection, while the r38 kDa antigen recorded a sensitivity of 73.8% and a specificity of 84.6% . In the case of PPD, the results were not promising . CONCLUSION: This paper on ELISA-based diagnostic tests attempts to implement an optimal decision support system through PRT that would identify the outcome (as infected or non-infected) based on the OD values . The PRT was able to predict the outcome for individual suspects . Further, Kullback-Leibler distance measurement has validated the PRT in distinguishing infected individuals from healthy subjects (based on the OD values).

Genome Biol . 2003;4(2):R8 . Epub 2003 Jan 30.
A search for doxycycline-dependent mutations that increase Drosophila melanogaster life span identifies the VhaSFD, Sugar baby, filamin, fwd and Cctl genes; Landis GN et al.; BACKGROUND: A P-type transposable element called PdL has been engineered with a doxycycline-inducible promoter directed out through the 3' end of the element . Insertion of PdL near the 5' end of a gene often yields doxycycline-dependent overexpression of that gene and a mutant phenotype . This functional genomics strategy allows for efficient screening of large numbers of genes for overexpression phenotypes . RESULTS: PdL was mobilized to around 10,000 new locations in the Drosophila melanogaster genome and used to search for genes that would extend life span when overexpressed . Six lines were identified in which there was a 5-17% increase in life span in the presence of doxyxcycline . The mutations were molecularly characterized and in each case a gene was found to be overexpressed using northern blots . Two genes did not have previously known phenotypes and are implicated in membrane transport: VhaSFD encodes a regulatory subunit of the vacuolar ATPase proton pump (H+-ATPase), whereas Sugar baby (Sug) is related to a maltose permease from Bacillus . Three PdL mutations identified previously characterized genes: filamin encodes the homolog of an actin-polymerizing protein that interacts with presenilins . four wheel drive (fwd) encodes a phosphatidylinositol-4-kinase (PI 4-kinase) and CTP:phosphocholine cytidylyltransferase-l (Cctl) encodes the rate-limiting enzyme in phosphatidylcholine synthesis . Finally, an apparently novel gene (Red herring, Rdh) was found in the first intron of the encore gene . CONCLUSIONS: Screening for conditional mutations that increase Drosophila life span has identified genes implicated in membrane transport, phospholipid metabolism and signaling, and actin cytoskeleton organization.

Biosci Biotechnol Biochem, 2003 Jan, 67(1), 94 - 9
Binding properties of Bacillus thuringiensis Cry1C delta-endotoxin to the midgut epithelial membranes of Culex pipiens; Kamauchi S et al.; The Cry1C delta-endotoxin from Bacillus thuringiensis is toxic to both lepidopteran and dipteran insect larvae . To analyze the dipteran-specific insecticidal mechanisms, we investigated the properties of Cry1C binding to the epithelial cell membrane of the larval midgut from the mosquito Culex pipiens in comparison with dipteran-specific Cry4A . Immunohistochemical staining of the larval midgut sections from Culex pipiens showed that Cry1C and Cry4A bound to the microvilli of the epithelial cells . The Cry1C binding to brush border membrane vesicles from the mosquito larvae was specific and irreversible, and did not compete with Cry4A . By ligand blotting analyses, we detected several Cry1C-binding proteins, the Cry1C binding to which did compete with excess unlabeled Cry4A . These results suggested that Cry1C and Cry4A recognized the same binding site(s) on the epithelial cell surface but that their interaction with the target membrane differed.

Bioresour Technol, 2003 Jul, 88(3), 251 - 4
Enhanced production of pectinase by Bacillus sp . DT7 using solid state fermentation; Raj Kashyap D et al.; Bacillus sp . DT7 produced very high levels of alkaline and thermotolerant pectinase by solid state fermentation . Production of this enzyme was affected by nature of solid substrate, level of moisture content, presence or absence of carbon, nitrogen, mineral and vitamin supplements . Maximum enzyme production of 8050 U/g dry substrate was obtained in wheat bran supplemented with polygalacturonic acid (PGA; 1%, w/v) and neurobion (a multivitamin additive; 27 micro l/g dry substrate) with distilled water at 75% moisture level, after 36 h of incubation at 37 degrees C.

Indian J Gastroenterol . 2003 Jan-Feb;22(1):25.
Mycobacterium avium-intracellulare in stool in HIV-seropositive man; Mirdha BR; Mycobacterium tuberculosis is the predominant acid-fast bacillus causing diarrhea in HIV-seropositive patients in India . We report a 27-year-old HIV-seropositive man with diarrhea in whom M . avium-intracellulare was isolated on stool culture.

Med Trop (Mars), 2002, 62(5), 473 - 9
{The Mycobacterium leprae genome: from sequence analysis to therapeutic implications}; Honore N; The genome of Mycobacterium leprae, the causative agent of leprosy, was analyzed by rapid sequencing of cosmids and plasmids prepared from DNA isolated from one patient's strain . Results showed that the bacillus possesses a single circular chromosome that differs from other known mycobacterium chromosomes with regard to size (3.2 Mb) and G + C content (57.8%) . Computer analysis demonstrated that only half of the sequence contains protein-coding genes . The other half contains pseudogenes and non-coding sequences . These findings indicate that M . leprae has undergone a major reductive evolution leaving a minimal set of functional genes for survival . Study of the coding region of the sequence provides evidence accounting for the particular pathogenic properties of M . leprae which is an obligate intracellular parasite . Disappearance of numerous enzymatic pathways in comparison with M . tuberculosis, an intracellular pathogen comparable to M . leprae, could explain the differences observed between the two organisms . Genomic analysis of the leprosy bacillus also provided insight into the molecular basis for resistance to various antibiotics and allowed identification of several potential targets for new drug treatments.

Vaccine, 2003 Mar 28, 21(13-14), 1518 - 26
Immune responses of elk to Mycobacterium bovis bacillus Calmette Guerin vaccination; Waters WR et al.; Although rare, detection of Mycobacterium bovis infection of captive or free-ranging elk (Cervus elaphus) elicits serious concern due to regulatory and zoonotic implications . Few studies, however, have evaluated the immune response of elk to M . bovis or other pathogens . To model natural infection, elk were vaccinated with live M . bovis bacillus Calmette Guerin (BCG, Pasteur strain) for evaluation of immune responsiveness to this attenuated live vaccine . Peripheral blood mononuclear cells (PBMC) of vaccinated elk proliferated in response to stimulation with a soluble mycobacterial antigen preparation (i.e . M . bovis purified protein derivative, PPDb) . Greater numbers of sIgM(+) cells (i.e . B cells) proliferated in this response than did either CD4(+), gammadeltaTCR(+) or CD8(+) cells . The in vivo response (i.e . delayed type hypersensitivity, DTH) to PPDb by vaccinated elk exceeded both the response by non-vaccinated elk and BCG-vaccinated cattle at 24, 48, and 72h post-administration of PPD . In vivo responses to PPDb by vaccinated elk diminished after 72h as compared to responses at 24 and 48h . Serum was also collected periodically and evaluated by ELISA for immunoglobulin (i.e . IgG heavy and light chains) reactivity to crude mycobacterial antigens . Two weeks post-vaccination and throughout the duration of the study, serum immunoglobulin reactivity to PPDb and to a proteinase K-digested whole cell sonicate of BCG exceeded that of serum from non-vaccinated elk . Intradermal administration of PPD for measurement of hypersensitive responses boosted the serum antibody response . These findings demonstrate that BCG vaccination of elk induces a serum antibody response to crude M . bovis antigens, a B cell in vitro proliferative response, and in vivo trafficking of mononuclear cells to sites of mycobacterial antigen administration (i.e . delayed type hypersensitivity) . A predominant B cell in vitro proliferative response by elk PBMC to crude mycobacterial test antigens will likely impact the development of improved diagnostic tests of tuberculosis infection for this species.

Vaccine, 2003 Mar 28, 21(13-14), 1502 - 9
Induction of specific T-helper and cytolytic responses to epitopes displayed on a virus-like protein scaffold derived from the pyruvate dehydrogenase multienzyme complex; Domingo GJ et al.; The icosahedral protein scaffold (1.5MDa) generated by self-assembly of the catalytic domains of the dihydrolipoyl acetyltransferase core of the pyruvate dehydrogenase multienzyme complex from Bacillus stearothermophilus has been engineered to display 60 copies of one or more peptide epitopes on a single molecule (E2DISP) . An E2DISP scaffold displaying pep23, a 15-residue B- and T-helper epitope from the reverse transcriptase of HIV-1, was able to induce a pep23-specific T-helper response in cell lines in vitro . The same scaffold displaying both pep23 and peptide RT2, a nine-residue CTL epitope from HIV-1 reverse transcriptase, was able to prime an RT2-specific CD8(+) T-cell response in human cell lines in vitro and in HLA-A2 transgenic mice in vivo . This was accompanied by a humoral antibody response specific for E2DISP-presented epitopes . Thus, the icosahedral acetyltransferase core constitutes a simple and flexible scaffold for multiple epitope display with access to both cellular and humoral immune response pathways.

Diagn Microbiol Infect Dis, 2003 Feb, 45(2), 143 - 7
"Bacillus hackensackii" sp . nov., a novel carbon dioxide sensitive bacterium isolated from blood culture; Hong T et al.; An endospore-forming, gram-positive bacillus was isolated from a patient's blood culture . This bacillus did not grow in the presence of 5% carbon dioxide although it grew well in ambient air at 37 degrees C . Although the organism thus is an aerobic bacterium, its sensitivity to increased carbon dioxide concentration places it in a distinct category of gaseous atmospheric requirement: capnophobic . Based on its morphology, growth characteristics, biochemical reactions and a complete 16S rRNA gene nucleotide sequence analysis, this microorganism represents a novel Bacillus species . The clinical significance of this isolate is unknown . It is proposed that the bacterium be classified in the genus Bacillus as "Bacillus hackensackii".

Clin Exp Allergy, 2003 Mar, 33(3), 386 - 93
Extended freeze-dried Mycobacterium bovis Bacillus Calmette-Guérin induces the release of interleukin-12 but not tumour necrosis factor-alpha by alveolar macrophages, both in vitro and in vivo; Hubeau C et al.; BACKGROUND: Airway hyper-responsiveness (AHR), chronic airway inflammation and predominance of the T helper type-2 (Th2; IL-4, IL-5, IL-13) over the Th1 (IL-2, IFN-gamma) immune response are hallmarks of asthma . Alveolar macrophages (AM) are the most numerous cells in the airway lumen, where they represent the first immune cell population encountered by inhaled antigens . AM act as antigen-presenting cells (APC) and they release various soluble mediators and enzymes . AM thus play a prominent role in the modulation of the local immunity in airways . In allergic airways, AM have been implicated in the pathogenesis of inflammation by promoting the Th2 versus the Th1 cytokine patterns . OBJECTIVES: Infections with attenuated bacteria or challenges with bacterial products may involve AM . Such stimuli have been shown to potentially restore the Th1/Th2 balance in asthmatic airways, but they also induce the release of inflammatory mediators . We investigated the response of AM when stimulated by two preparations of non-proliferating Bacillus Calmette-Guerin (BCG) . METHODS: We evaluated the cytokine production by AM from BP2 and C57BL/6 mice when cultured with heat-killed (HK) and extended freeze-dried (EFD) BCG . We then investigated in vivo the release of soluble factors in the airway lumen of mice after instillation of these BCG preparations . Finally, we studied the profile of cytokine transcripts in the lung of mice pre-treated with BCG and then challenged with ovalbumin (OVA) . RESULTS: HK BCG induced the production of both TNF-alpha and IL-12, and did not prevent high levels of Th2 cytokine transcripts . In contrast, EFD BCG induced a response dominated by the production of IL-12, with no later over-expression of Th2 cytokine transcripts . CONCLUSION: Our results show that EFD BCG induce the release of the Th1-promoting cytokine IL-12 by AM, without the deleterious effects of HK BCG . These data suggest that EFD BCG may be considered as a potential novel treatment to restore the Th1/Th2 imbalance in asthma.

Probl Tuberk, 2002, (12), 3 - 7
{Organizational and methodological approaches to improving the microbiological diagnosis of tuberculosis in Russia}; Golyshevskaia VI et al.; The practice of the researchers of the Central Research Institute of Tuberculosis in Russia's experimental areas yielded approaches to improving the microbiological diagnosis of tuberculosis in both specialized and general health care facilities . A decisive role in enhancing the efficiency of microbiological identification of patients with tuberculosis is played by the following factors: laboratory staff training, provision of laboratories with qualitative equipment and reagents, introduction of unified methods and those approved by the Ministry of Health of the Russian Federation, regular supervision of jurisdictional laboratories, and outside test quality control . The efficiency of microbiological diagnosis of bacillary patients primarily depends on the expediency of formation of groups of patients under study, the observance of the frequency of examinations, and the assurance of the quality of collected materials . With this, the centralization of microbiological tests for tuberculosis at the most trained, certified, and licensed large laboratories that exercise a centralized control over the quality of microbiological tests.

J Clin Oncol, 2003 Mar 1, 21(5), 884 - 90
Phase II trial of autologous tumor vaccination, anti-CD3-activated vaccine-primed lymphocytes, and interleukin-2 in stage IV renal cell cancer; Chang AE et al.; PURPOSE: Previous preclinical and clinical studies have demonstrated that autologous tumor vaccines can induce relatively specific tumor-reactive T cells in draining lymph nodes . The adoptive transfer of these cells can result in tumor regression . PATIENTS AND METHODS: Patients with stage IV renal cell cancer (RCC) were vaccinated with irradiated autologous tumor cells admixed with Calmette-Guerin bacillus . Approximately 7 days later, vaccine-primed lymph nodes (VPLNs) were harvested and the lymphoid cells secondarily activated with anti-CD3 monoclonal antibody and expanded in interleukin 2 (IL-2) . The activated cells were subsequently infused intravenously along with the concomitant administration of bolus IL-2 (360,000 U/kg intravenously x 15 doses) . RESULTS: Thirty-nine patients were entered onto the study, of whom 34 completed an initial course of cell therapy consisting of a mean (SEM) number of 4.3 (2.2) x 10(10) VPLN cells . Among subjects who received cell therapy, there were nine responses (four complete responses {CRs} and five partial responses {PRs}), for an overall response rate of 27% . The durations of the CRs were > 48, 45, > 35, and 12 months, and the durations of the PRs were > 63, 48, 15, 12, and 4 months . Cultured tumor cells were available to assess in vitro cytokine release of VPLN cells in 24 subjects . The median cytokine release ratio of interferon gamma (IFNgamma) to IL-10 for responders and nonresponders was 992 and 5, respectively, which was significantly different (P =.047) . CONCLUSION: The treatment protocol resulted in durable tumor responses in patients with advanced RCC . The ratio of IFNgamma and IL-10 cytokines released in response to tumor by the VPLN cells was a significant correlate with tumor response.

Shokuhin Eiseigaku Zasshi, 2002 Oct, 43(5), 273 - 9
Effect of subchronic feeding of genetically modified corn (CBH351) on immune system in BN rats and B10A mice; Teshima R et al.; Subchronic animal feeding studies to examine the effect on the immune system of genetically modified corn CBH351, which contains the Cry9C protein derived from Bacillus thuringiensis subspecies tolworthi, were conducted in female BN rats and B10A mice . The studies were designed to compare the effect of a line of genetically modified corn CBH351 (GM corn) with that of isoline corn (non-GM corn) . Heat-treated corn meal was incorporated into the diets of the rats and mice at a concentration of 50% . The study duration was 13 weeks . Growth, food intake, and organ weights of the thymus, spleen, and liver were compared between animals fed the non-GM and GM lines . The histological findings in thymus, spleen, mesenteric lymph nodes, Peyer's patches, small intestines, liver, kidney, and bone marrow, and the presence of Cry9C-specific IgE, IgG, IgG1 and IgA antibodies in serum were also compared . The results showed no significant differences in growth, feeding value, or the histological findings in immunity-related organs between the animals fed the GM and non-GM lines . Production of Cry9 C-specific IgE and IgA was not detected in the serum of either group . Production of Cry9C-specific IgG and IgG1 was slightly increased in the 50% GM groups of BN rats . No Cry9C-specific IgG or IgG1 was detected in the serum of BN rats fed the diet containing 5% GM-corn In conclusion, no immunotoxic activity was detected in the GM-corn-fed rats and mice in this subchronic dietary study.

Med Hypotheses, 2003 Feb, 60(2), 290 - 2
Is this the historical 'cancer germ'?
Wainwright M, Al Talih A.
A highly pleomorphic bacterium, isolated from a canine mammary tumour was found to share many characteristics with recently described cancer-related bacteria and with the Glover organism, a historically important 'cancer germ' . We suggest that both the Glover organism, and possibly other cancer-related bacteria, are likely to be a strain of the highly pleomorphic bacterium, Bacillus licheniformis.

Am J Hematol, 2003 Mar, 72(3), 204 - 8
Fatal Bacillus cereus sepsis following resolving neutropenic enterocolitis during the treatment of acute leukemia; Ginsburg AS et al.; Bacillus cereus is increasingly being acknowledged as a serious bacterial pathogen in immunosuppressed hosts . We report a case of fatal B . cereus sepsis in a patient with newly diagnosed acute leukemia following resolving neutropenic enterocolitis . Gastrointestinal complaints are common during induction chemotherapy, yet some antimicrobial coverage suitable for generalized neutropenia is not optimal for the eradication of B . cereus . This case demonstrates that, in the neutropenic patient with gastrointestinal complaints or in the setting of resolving neutropenic enterocolitis, it is important to anticipate possible B . cereus infection and sepsis .

Arch Med Res, 2003 Jan-Feb, 34(1), 20 - 5
Transforming growth factor-beta decreases survival of Mycobacterium bovis-activated T cells; Hernandez-Garay M et al.; BACKGROUND: A comprehensive understanding of the immune response induced by Mycobacterium bovis Bacille Calmette-Guerin in activation of protective T cells against tuberculosis is important to develop effective therapies to combat this disease . In this study, our experiments were designed to determine effects of transforming growth factor (TGF)-beta on M . bovis-induced T-cell activation and survival . METHODS: Fluorescence-activated cell sorter (FACS) analysis was used for detection of apo-ptotic cells by three different methods: 1) . scattered light change during early phase of apoptosis; 2) . detection of hypodiploid DNA, or 3) . terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) technique . Quantification of positively stained population was based on samples stained with isotype control antibodies analyzed on a FACScan . RESULTS: TGF-beta added at initiation of culture did not alter percentage of viable cells . By contrast, TGF-beta added 72 h post-activation decreased percentage of viable cells . This effect was statistically significant (p <0.05) . Furthermore, addition of anti-TGF-beta MoAb together with TGF-beta abolished the ability of this cytokine to decrease survival in post-activated human T cells . Role of TGF-beta on post-activated human T cells was further confirmed by staining apoptotic nuclei with propidium iodide, which detects late events of apoptosis, and by DNA fragmentation determined using TUNEL assay . Interestingly, TGF-beta did not promote Fas-mediated killing . Finally, TGF-beta increased apoptosis of CD4(+) T cells after mycobacterial stimulation . CONCLUSIONS: This study indicated an important role for TGF-beta in suppression of protective immune response against M . bovis by promoting elimination of post-activated T cells . Furthermore, results showed that TGF-beta had no direct effect on M . bovis-induced up-regulation of Fas (CD95).

Immunology, 2003 Mar, 108(3), 352 - 64
Dendritic cells recruited to the lung shortly after intranasal delivery of Mycobacterium bovis BCG drive the primary immune response towards a type 1 cytokine production; Lagranderie M et al.; We showed in a previous study that the intranasal (i.n) delivery of bacille Calmette-Guerin (BCG) to BP2 mice (H-2q) inhibits eosinophilia and bronchial hyperreactivity in a mouse model of asthma . The present work has been performed to characterize the leucocyte lineages recruited to the lungs of mice after i.n . delivery of BCG and potentially involved in the polarization of T lymphocytes . The different antigen-presenting cells (APC) recruited to bronchoalveolar lavage (BAL) and to lung tissue of mice shortly after the delivery of BCG were analysed in parallel as well as their capacity to drive the immune response towards a T helper type 1 cytokine production . Alveolar macrophages (AM) from the BAL were CD11c+, F4/80+ and CD11b-, and in the lung tissue two major populations of potential APC were detected: one CD11c-, F4/80+, CD11b+ and I-Aq- was identified as interstitial macrophages (IM) and a second expressing CD11c+ and I-Aq+ antigens, negative for CD11b and F4/80 markers as leucocytic dendritic cells (DC) . Freshly isolated DC up-regulated CD11b and CD40 antigens after overnight culture, but remained negative for CD8alpha antigen, suggesting a myeloid origin . Lung DC which produced high amount of interleukin (IL)-12 were potent inducers of naive CD4+ T lymphocyte priming, as assessed by interferon-gamma (IFN-gamma) production by these naive CD4+ T cells . Lung explants recovered long term after BCG delivery produced sustained levels of IFN-gamma . Our results suggest that AM and particularly DC by secreting IL-12 shortly after BCG delivery induce the long-term persistence of IFN-gamma-secreting T cells percolating in BCG-loaded lung tissue.

Eur J Biochem, 2003 Mar, 270(5), 861 - 70
Site-directed mutagenesis of a loop at the active site of E1 (alpha2beta2) of the pyruvate dehydrogenase complex . A possible common sequence motif; Fries M et al.; Limited proteolysis of the pyruvate decarboxylase (E1, alpha2beta2) component of the pyruvate dehydrogenase (PDH) multienzyme complex of Bacillus stearothermophilus has indicated the importance for catalysis of a site (Tyr281-Arg282) in the E1alpha subunit (Chauhan, H.J., Domingo, G.J., Jung, H.-I . & Perham, R.N . (2000) Eur . J . Biochem . 267, 7158-7169) . This site appears to be conserved in the alpha-subunit of heterotetrameric E1s and multiple sequence alignments suggest that there are additional conserved amino-acid residues in this region, part of a common pattern with the consensus sequence -YR-H-D-YR-DE- . This region lies about 50 amino acids on the C-terminal side of a 30-residue motif previously recognized as involved in binding thiamin diphosphate (ThDP) in all ThDP-dependent enzymes . The role of individual residues in this set of conserved amino acids in the E1alpha chain was investigated by means of site-directed mutagenesis . We propose that particular residues are involved in: (a) binding the 2-oxo acid substrate, (b) decarboxylation of the 2-oxo acid and reductive acetylation of the tethered lipoyl domain in the PDH complex, (c) an "open-close" mechanism of the active site, and (d) phosphorylation by the E1-specific kinase (in eukaryotic PDH and branched chain 2-oxo acid dehydrogenase complexes).

J Eur Acad Dermatol Venereol, 2003 Jan, 17(1), 65 - 7
A case of bacillary angiomatosis presenting as leg ulcers; Karakas M et al.; We report a 32-year-old immunocompetent man who had multiple leg ulcers caused by bacillary angiomatosis without a history of direct contact with cats . Bacillary angiomatosis should be kept in mind in the differential diagnosis of leg ulcers in cases of unknown aetiology.

J Environ Biol, 2002 Apr, 23(2), 111 - 35
Host plant resistance to insects: an eco-friendly approach for pest management and environment conservation; Sharma HC et al.; Host plant resistance (HPR) to insects is an effective, economical, and environment friendly method of pest control . The most attractive feature of HPR is that farmers virtually do not need any skill in application techniques, and there is no cash investment by the resource poor farmers . Considerable progress has been made in identification and development of crop cultivars with resistance to the major pests in different crops . There is a need to transfer resistance genes into high-yielding cultivars with adaptation to different agro-ecosystems . Resistance to insects should form one of the criteria to release varieties and hybrids for cultivation by the farmers . Genes from the wild relatives of crops, and novel genes, such as those from Bacillus thuringiensis can also be deployed in different crops to make HPR an effective weapon to minimize the losses due to insect pests . HPR will not only cause a major reduction in pesticide use and slowdown the rate of development of resistance to insecticides in insect populations, but also lead to increased activity of beneficial organisms and reduction in pesticide residues in food and food products.

Rev Prat, 2002 Dec 1, 52(19), 2133 - 8
{Tuberculosis in children}; Gaudelus J; Children tuberculosis accounts for 5.5% of the declared cases of tuberculosis in France . Tuberculosis in children is always due to recent contamination from an adult . Survey of tuberculosis in children is a good indicator of the Koch's bacillus spread . Age is an important risk factor for an infected child to develop tuberculous disease . Estimated risk is 43% under 1 year of age, 24% between 1 and 5 years of age, 16% in teenagers . The risk to develop severe disease (miliary, meningitis) is higher in infants . Teenagers may have a rapidly evolutive form . Diagnosis of tuberculosis in children is difficult . One has to keep in mind this diagnosis which must be assessed by tuberculin skin test, radiologic exams and bacteriological sampling . The proof of tuberculosis is rare in children: direct examination of gastric fluid is positive only in 20% of cases or less; culture is positive only in 50% of cases or less . Infection and disease have to be treated in children . The search of the index case is mandatory.

Mol Cell Proteomics, 2003 Jan, 2(1), 19 - 28
Insect resistance to Bacillus thuringiensis: alterations in the indianmeal moth larval gut proteome; Candas M et al.; Insect resistance to the Cry toxins of Bacillus thuringiensis (Bt) has been examined previously using a number of traditional biochemical and molecular techniques . In this study, we utilized a proteomic approach involving two-dimensional differential gel electrophoresis, mass spectrometry, and function-based activity profiling to examine changes in the gut proteins from the larvae of an Indianmeal moth (IMM, Plodia interpunctella) colony exhibiting resistance to Bt . We found a number of changes in the levels of certain specific midgut proteins that indicate increased glutathione utilization, elevation in oxidative metabolism, and differential maintenance of energy balance within the midgut epithelial cells of the Bt-resistant IMM larva . Additionally, the electrophoretic migration pattern of a low molecular mass acidic protein, which apparently is an ortholog of F(1)F(0)-ATPase, was considerably altered in the Bt-resistant insect indicating that variations in amino acid content or modifications of certain proteins also are important components of the resistance phenomenon in the IMM . Furthermore, there was a dramatic decrease in the level of chymotrypsin-like proteinase in the midgut of the Bt-resistant larva, signifying that reduction of chymotrypsin activity, and subsequently decreased activation of Cry toxin in the insect midgut, is an important factor in the resistant state of the IMM . The proteomic analysis of larval gut proteins utilized in this study provides a useful approach for consolidating protein changes and physiological events associated with insect resistance to Bt . Our results support the hypothesis that physiological adaptation of insects and resistance to Bt is multifaceted, including protein modification and changes in the synthesis of specific larval gut proteins . We believe that increased oxidative metabolism may be an adaptive response of insects that undergo survival challenge and that it could mediate detoxification as well as higher rates of generalized and localized mutations that enhance their resistance and provide survival advantage.

Rev Argent Microbiol, 2002 Oct-Dec, 34(4), 230 - 4
{Activity of 14 antimicrobials against Eikenella corrodens}; Vay C et al.; Eikenella corrodens is a gram-negative bacillus that colonizes as normal flora of the mouth, the upper respiratory tract and the gastrointestinal tract . The aim of this study was to determine the susceptibility patterns against fourteen antibiotics of 25 E . corrodens strains isolated at our hospital . MICs were determined by the agar dilution technique using Mueller-Hinton agar with sheep blood (5% v/v) to penicillin, ampicillin, ampicillin-sulbactam, cephalotin, cefoxitin, ceftiaxone, colistin, gentamicin, amikacin, erythromycin, rifampin, ciprofloxacin and clindamycin . The most active antibiotics were ciprofloxacin and ceftriaxone (MIC90 = 0.008 and 0.125 microgram/ml, respectively), whereas eritromycin, gentamicin and amikacin showed less activity . Only one strain was beta lactamase positive, and it was inhibited by sulbactam . Erithromycin, gentamicin and amikacin had poor activity (MIC90 = 16.8 and 64 micrograms/ml, respectively), whereas all the strains were uniformly resistant to clindamycin (MIC > or = 32 micrograms/ml) . We suggest about the need of periodical surveys of E . corrodens susceptibility patterns, since strains have been found with decreased susceptibility against antibiotics which are currently being used for the treatment of infectious diseases.

Hua Xi Yi Ke Da Xue Xue Bao, 2002 Jan, 33(1), 35 - 9
{Construction and identification of recombinant shuttle-plasmid with ESAT-6 from Mycobacterium tuberculosis}; Chen W et al.; OBJECTIVE: To construct a recombinant BCG secretively expressing ESAT-6 of Mycobacterium tuberculosis . METHODS: alpha-antigen(alpha-Ag) signal sequence and esat-6 gene were amplified from the genome of Bacille Calmette-Guerin (BCG) and Mycobacterium tuberculosis by PCR respectively . esat-6 gene was cloned in E . coli-BCG shuttle-plasmid pMV261 to get pME . Then a new recombinant plasmid pSME was constructed by inserting BCG alpha-Ag signal sequence into pME . RESULTS: The cloned genes alpha-Ag signal sequence and esat-6 were correctly inserted into the vector pMV261, which was confirmed by restriction endonuclease digestion and PCR amplification of pSME . CONCLUSION: pSME was expected to secretively express ESAT-6 of Mycobacterium tuberculosis in BCG . This study provides the possibility of further researches on the development of new anti-tuberculosis vaccine.

Head Neck, 2003 Mar, 25(3), 198 - 209
Generation of vaccine-primed lymphocytes for the treatment of head and neck cancer; Chang AE et al.; BACKGROUND: This study was performed to assess the ability of autologous tumor vaccines to induce T-cell reactivity to squamous cell cancers (SCC) . METHODS: Irradiated autologous tumor cells admixed with bacillus Calmette-Guerin (BCG) were given intradermally in patients with advanced head and neck cancers . Vaccine-primed lymph node (VPLN) cells were secondarily activated with anti-CD3 mAb and expanded in IL-2 for adoptive immunotherapy . A mean (+/- SEM) of 2 (+/-0.6) x 10(10) anti-CD3-activated cells were administered in conjunction with IL-2 in six patients . RESULTS: Anti-CD3-activated VPLN cells secreted IFN-gamma and GM-CSF in response to autologous tumor cells but not to allogeneic tumor cells in four of five patients analyzed . Both CD4(+) and CD8(+) tumor reactive cells were present in the VPLN . There were no significant tumor responses after transfer of the anti-CD3-activated VPLN . In separate experiments, costimulation of VPLN cells with anti-CD3 and anti-CD28 mAb resulted in enhanced cytokine secretion to autologous tumor compared with anti-CD3 activation alone . CONCLUSIONS: Both CD4(+) and CD8(+) responses can be induced to SCC by autologous tumor vaccination . However, additional approaches need to be identified to enhance the therapeutic efficacy of this approach .

Proc Natl Acad Sci U S A, 2003 Mar 4, 100(5), 2760 - 5 Epub 2003 Feb 21.
Bacillus thuringiensis crystal proteins that target nematodes; Wei JZ et al.; Bacillus thuringiensis (Bt) crystal proteins are pore-forming toxins used as insecticides around the world . Previously, the extent to which these proteins might also target the invertebrate phylum Nematoda has been mostly ignored . We have expressed seven different crystal toxin proteins from two largely unstudied Bt crystal protein subfamilies . By assaying their toxicity on diverse free-living nematode species, we demonstrate that four of these crystal proteins are active against multiple nematode species and that each nematode species tested is susceptible to at least one toxin . We also demonstrate that a rat intestinal nematode is susceptible to some of the nematicidal crystal proteins, indicating these may hold promise in controlling vertebrate-parasitic nematodes . Toxicity in nematodes correlates with damage to the intestine, consistent with the mechanism of crystal toxin action in insects . Structure-function analyses indicate that one novel nematicidal crystal protein can be engineered to a small 43-kDa active core . These data demonstrate that at least two Bt crystal protein subfamilies contain nematicidal toxins.

J Biol Chem, 2003 May 2, 278(18), 15713 - 9 Epub 2003 Feb 20.
DNA (cytosine-N4-)- and -(adenine-N6-)-methyltransferases have different kinetic mechanisms but the same reaction route . A comparison of M.BamHI and T4 Dam; Malygin EG et al.; We studied the kinetics of methyl group transfer by the BamHI DNA-(cytosine-N(4)-)-methyltransferase (MTase) from Bacillus amyloliquefaciens to a 20-mer oligodeoxynucleotide duplex containing the palindromic recognition site GGATCC . Under steady state conditions the BamHI MTase displayed a simple kinetic behavior toward the 20-mer duplex . There was no apparent substrate inhibition at concentrations much higher than the K(m) for either DNA (100-fold higher) or S-adenosyl-l-methionine (AdoMet) (20-fold higher); this indicates that dead-end complexes did not form in the course of the methylation reaction . The DNA methylation rate was analyzed as a function of both substrate and product concentrations . It was found to exhibit product inhibition patterns consistent with a steady state random bi-bi mechanism in which the dominant order of substrate binding and product release (methylated DNA, DNA(Me), and S-adenosyl-l-homocysteine, AdoHcy) was Ado-Met DNA DNA(Me) AdoHcy . The M.BamHI kinetic scheme was compared with that for the T4 Dam (adenine-N(6)-)-MTase . The two differed with respect to an effector action of substrates and in the rate-limiting step of the reaction (product inhibition patterns are the same for the both MTases) . From this we conclude that the common chemical step in the methylation reaction, methyl transfer from AdoMet to a free exocyclic amino group, is not sufficient to dictate a common kinetic scheme even though both MTases follow the same reaction route.

Life Sci Space Res, 1977, 15, 65 - 72
Morphology of extremely heat-resistant spores from Bacillus sp . ATCC 27380 by scanning and transmission electron microscopy; Youvan D et al.; Bacillus sp . ATCC 27380 is a recently discovered aerobic mesophile, isolated from surface soil, that produces spores with extreme resistance to dry heat: the length of time to 90% kill is 139 hr at 125 degrees C and 13-17 hr at 138 degrees C . Values for spores from other known species range from 5 to 100 min . The molecular basis for this extreme heat resistance is unknown . We report a structural analysis of the internal and external mature spore morphology obtained by both scanning and transmission electron microscopy . Both modes of microscopy delineate a morel-like structure characterized by irregular, but distinct, polygonal ridges suggestive of extreme dehydration . Some spores also possess an appendage resembling the bun of a brioche . This bun-like body is possibly unique to this species . In cross section the spore exhibits a many-layered structure, each layer with a characteristic fine structure . These morphological characters do not suffice to explain the observed resistance to dry heat at extreme temperatures . They do form a basis for the chemical characterizations which will be necessary to understand this heat resistance at the molecular level . The concept of "solid state spore" is put forward as a generalization that may be useful towards understanding this resistance.

Life Sci Space Res, 1977, 15, 47 - 52
Physiology of xerophytic micro-organisms growing under Martian conditions; Imshenetsky AA et al.; The main factor limiting growth of terrestrial organisms under Martian conditions is the low humidity; therefore the biology of xerophytic micro-organisms has been studied . We found earlier that Bacillus megaterium var . halotolerant and Mycococcus ruber could survive and grow under Martian conditions . In this work, we studied the zone of tolerance of these and some other micro-organisms towards various levels of water activity, and the toxicity of substances which were used to control this activity . A technique for quantitative evaluation of the xerophytic nature of micro-organisms is proposed . The technique is based on quantitative evaluation by gas chromatography of carbon dioxide evolved in the course of growth of micro-organisms on media with different values of water activity.

Bull Soc Pathol Exot, 2002 Nov, 95(4), 265 - 8
{Preliminary study on isoniazid-epiroprim combination in a tuberculosis murine model}; N'guessan K et al.; The purpose of this study regarding isoniazid-epiroprim's association applied to antituberculosis chemotherapy, carried through murine model, initiated into Institut Pasteur of Cote d'Ivoire and worked out at Institut Pasteur of Paris was to evaluate the epiroprim's effect alone and associated with isoniazid on Mycobacterium tuberculosis . Sixteen mouses (lineage C57Bl/6) were inoculated by venous way with 10(5) viable bacillus (strain H37Rv) suspended in 500 microliters sterile physiological aqueous solution and were shared out into 4 sets . Fifteen days later the sets have been submitted or not to a daily treatment by gavage during three weeks (epiroprim, isoniazid, isoniazid plus epiroprim) . The mouses were euthanasied, spleen and lung were removed from each animal . The titres of determined bacillus into those organs prove that isoniazid and epiroprim associated seem more efficacious than the isoniazid monotherapy for mouses pulmonary tuberculosis . Bacillus obtained are sensitive to isoniazid.

Acta Crystallogr D Biol Crystallogr, 2003 Mar, 59(Pt 3), 591 - 4 Epub 2003 Feb 21.
Crystallization and preliminary X-ray diffraction studies of a mosquito-larvicidal toxin from Bacillus thuringiensis subsp . israelensis; Boonserm P et al.; The Cry4B delta-endotoxin from Bacillus thuringiensis subsp . israelensis is specifically toxic to mosquito larvae . For a better understanding of the mechanism of toxicity, chymotrypsin-activated Cry4B toxin (68 kDa) has been purified and crystallized in sodium bromide at neutral pH . The well formed crystals belong to the rhombohedral space group R32, with unit-cell parameters a = b = 185.82, c = 187.93 A, and diffracted X-rays to 1.75 A resolution . The asymmetric unit contains one toxin molecule and 74% solvent content, as shown by molecular replacement from a composite model of the homologous Cry3A and Cry1Aa . The purified protein and crystals both possessed mosquitocidal activity.

Acta Crystallogr D Biol Crystallogr, 2003 Mar, 59(Pt 3), 447 - 52 Epub 2003 Feb 21.
Direct experimental observation of the hydrogen-bonding network of a glycosidase along its reaction coordinate revealed by atomic resolution analyses of endoglucanase Cel5A; Varrot A et al.; Non-covalent interactions between protein and ligand at the active centre of glycosidases play an enormous role in catalysis . Dissection of these hydrogen-bonding networks is not merely important for an understanding of enzymatic catalysis, but is also increasingly relevant for the design of transition-state mimics, whose tautomeric state, hydrogen-bonding interactions and protonation contribute to tight binding . Here, atomic resolution ( approximately 1 A) analysis of a series of complexes of the 34 kDa catalytic core domain of the Bacillus agaradhaerens endoglucanase Cel5A is presented . Cel5A is a 'retaining' endoglucanase which performs catalysis via the formation and subsequent breakdown of a covalent glycosyl-enzyme intermediate via oxocarbenium-ion-like transition states . Previous medium-resolution analyses of a series of enzymatic snapshots has revealed conformational changes in the substrate along the reaction coordinate (Davies et al., 1998) . Here, atomic resolution analyses of the series of complexes along the pathway are presented, including the 'Michaelis' complex of the unhydrolysed substrate, the covalent glycosyl-enzyme intermediate and the complex with the reaction product, cellotriose . These structures reveal intimate details of the protein-ligand interactions, including most of the carbohydrate-associated H atoms and the tautomeric state of crucial active-centre groups in the pH 5 orthorhombic crystal form and serve to illustrate the potential for atomic resolution analyses to inform strategies for enzyme inhibition.

J Mol Biol, 2003 Mar 7, 326(5), 1513 - 22
P but not R-axis interface is involved in cooperative binding of NAD on tetrameric phosphorylating glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus; Roitel O et al.; Homotetrameric phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus can be described as a dimer of dimers with three non-equivalent P, R, and Q interfaces . In our previous study, negative cooperativity in NAD binding to wild-type GAPDH was interpreted according to the induced-fit model in terms of two independent dimers with two interacting binding sites in each dimer . Two dimeric mutant GAPDHs, i.e . Y46G/S48G and D186G/E276G, were shown to exhibit positive cooperativity in NAD binding . Based on the molecular modeling of the substitutions and the fact that the most extensive inter-subunit interactions are formed across the P-axis interface of the tetramer, it was postulated that both dimeric mutant GAPDHs were of O-P type . Therefore, the P-axis interface was assumed to play a major role in causing cooperativity in NAD binding.Here, two other mutant GAPDHs, Y46G/R52G and D282G, have been studied . Using small angle X-ray scattering, the dimeric form of the D282G mutant GAPDH is shown to be of O-R type whereas both dimeric mutant GAPDHs Y46G/R52G and Y46G/S48G are of O-P type . Similarly to dimeric Y46G/S48G mutant GAPDH, the dimeric Y46G/R52G mutant GAPDH exhibits positive cooperativity in NAD binding . On the other hand, no significant cooperativity in NAD binding to the dimeric form of the D282G mutant GAPDH is observed, whereas its tetrameric counterpart exhibits negative cooperativity, similarly to the wild-type enzyme . Altogether, the results support the view that the P-axis interface is essential in causing cooperativity in NAD binding by transmitting the structural information induced upon cofactor binding from one subunit to the other one within O-P/Q-R dimers in contrast to the R-axis interface, which does not transmit structural information within O-R/Q-P dimers . The absence of activity of O-P and O-R dimer GAPDHs is the consequence of a pertubation of the conformation of the active site, at least of the nicotinamide subsite, as evidenced by the absence of an ion pair between catalytic residues C149 and H176 and the greater accessibility of C149 to a thiol kinetic probe.

Int J Food Microbiol, 2003 May 15, 82(3), 223 - 32
Contamination flows of Bacillus cereus and spore-forming aerobic bacteria in a cooked, pasteurized and chilled zucchini purée processing line; Guinebretiere MH et al.; A food processing plant producing pasteurized purees and its zucchini puree processing line were examined for contamination with aerobic and facultative anaerobic bacterial spores during a day's operation . Multiplication of spores was also monitored in the product stored under different conditions . High concentrations of Bacillus cereus spores were found in the soil in which the zucchinis were grown (4.6+/-0.3 log CFU/g), with a background spore population of 6.1+/-0.2 log CFU/g . In the processing plant, no B . cereus or psychrotrophic bacterial spores were detected on equipment . B . cereus and psychrotrophic bacterial spores were detected after enrichment in all samples of raw zucchinis, washed zucchinis, of two ingredients (starch and milk proteins) and in processed puree at each processing step . Steam cooking of raw zucchinis and pasteurization of puree in the final package significantly reduced spore numbers to 0.5+/-0.3 log CFU/g in the processed food . During storage, numbers of spore-forming bacteria increased up to 7.8+/-0.1 log CFU/g in puree after 5 days at 20-25 degrees C, 7.5+/-0.3 log CFU/g after 21 days at 10 degrees C and 3.8+/-1.1 log CFU/g after 21 days at 4 degrees C . B . cereus counts reached 6.4+/-0.5 log CFU/g at 20-25 degrees C, 4.6+/-1.9 log CFU/g at 10 degrees C, and remained below the detection threshold (1.7 log CFU/g) at 4 degrees C . Our findings indicate that raw vegetables and texturing agents such as milk proteins and starch, in spite of their low levels of contamination with bacterial spores and the heat treatments they undergo, may significantly contribute to the final contamination of cooked chilled foods . This contamination resulted in growth of B . cereus and psychrotrophic bacterial spores during storage of vegetable puree . Ways to eliminate such contamination in the processing line are discussed.

Indian J Med Res, 2002 Aug, 116, 64 - 9
BCG vaccination reaction in low birth weight infants; Kaur S et al.; BACKGROUND & OBJECTIVES: About 30 per cent newborns (preterm and term) weigh < 2500 g at birth . The immunological system is less mature in low birth weight (LBW) babies compared to term and normal birth weight (NBW) babies . Bacille Calmettee Guerin (BCG) vaccine is given at birth under the national immunization programme . There is a paucity of information on the immunogenicity of BCG vaccine in preterm and LBW babies . It was, therefore, proposed to study the reaction of BCG vaccination in LBW, preterm and normal birth weight newborns . METHODS: A total of 143 newborns (90 term and 53 preterm; of these 78 were LBW) received during March to September 1998, 0.1 ml of BCG vaccine (Danish 1331 strain) intradermally on the left arm just above the insertion of the deltoid muscle within 7 days of life . At the same time trivalent oral polio vaccine was administered as per the national immunization programme . These babies were followed up in the immunization clinic at 4, 6, 8, 10 and 12 +/- 1 wk to observe reactions at the BCG vaccination site . RESULTS: After 4 wk reaction at the vaccination site was significantly (P < 0.001) delayed in preterm babies as compared to term infants, and in the LBW babies (P < 0.05) as compared to NBW babies . The reaction at the site of vaccination was not found to be different at 6, 8, 10, 12 wk . BCG scar was seen in 47.5 per cent infants (45.4% in < 2500 g birth weight and 50% in > or = 2500 g birth weight infants) at 12 wk . But 33 (42.3%) LBW and 24 (36.9%) NBW infants also showed papule, pustule, ulceration or scab at the BCG vaccination site . INTERPRETATION & CONCLUSION: The BCG reaction was seen in the sequential order from papule to scar formation . No significant difference was seen in the scar formation in infants studied with varying gestation and birth weights after 12 wk of BCG vaccination . Fifty seven (40.4%) babies still showed different stages of BCG reaction at 12 wk . BCG vaccine along with OPV administered in early neonatal life showed successful BCG reaction in 95.5 per cent infants.

Scand J Immunol, 2003 Feb, 57(2), 151 - 61
T- and B-cell epitopes in the secreted Mycobacterium bovis antigen MPB70 in mice; Tollefsen S et al.; MPB70 is a soluble secreted protein highly expressed in Mycobacterium bovis and strains of bacille Calmette-Guerin (BCG); as such, it is a candidate for subunit and DNA vaccines against tuberculosis . MPB70 was screened for T-cell epitopes in four different inbred mouse strains . Major histocompatibility complex (MHC) H-2b-expressing mice (C57BL/6) secreted interferon-gamma (IFN-gamma) after stimulation with peptides from the regions 1-20, 41-50, 81-110, 121-150 and 161-193 of the MPB70 sequence . H-2db mouse (B6D2) splenocytes secreted IFN-gamma after stimulation with some of the same peptides, whereas H-2d mice (BALB/c and DBA/2) did not secrete IFN-gamma upon stimulation with the peptides . Sera from H-2db mice immunized with native MPB70 in incomplete Freund's adjuvant (IFA), mpb70 DNA or live BCG Moreau were found to contain antibodies against the native MPB70 antigen . H-2db mice immunized with native MPB70 in IFA exhibited high titres of peptide-reactive immunoglobulin G1 (IgG1) antibodies, whereas DNA-immunized mice reacted with IgG2a antibodies against some of the same peptides . As some of the epitopes recognized by mouse T and B cells have previously been found to stimulate immune responses in humans, cattle and rabbits, we conclude that these epitopes may be good general epitopes for the stimulation of T- and B-cell responses and candidates for a DNA vaccine with a broad applicability.

Scand J Immunol, 2003 Feb, 57(2), 107 - 14
Transfer of macrophage-derived mycobacterial antigens to dendritic cells can induce naïve T-cell activation; Girvan A et al.; Mycobacteria are capable of surviving and replicating in host macrophages, where they can release antigenic material into the environment . However, unlike dendritic cells (DCs), macrophages do not appear to be capable of activating naive T cells . Therefore, this work investigated antigen transfer between macrophages and DCs . We generated culture supernatants from bacille Calmette-Guerin (BCG)-infected and uninfected macrophages and then determined whether DCs could present these extracellular mycobacterial antigens to T cells . Here, we show that DCs pulsed with antigens released from BCG-infected macrophages can stimulate primed T cells in vitro and initiate naive T-cell responses in vivo . These results suggest that antigen transfer can occur between macrophages and DCs.

J Appl Microbiol, 2003, 94(3), 369 - 74
Identification of polyhydroxyalkanoate (PHA)-producing Bacillus spp . using the polymerase chain reaction (PCR); Shamala TR et al.; AIMS: The aim of the work was to develop efficient method to identify polyhydroxyalkanoate (PHA)-producing species of Bacillus from numerous soil isolates of bacteria . Identification of the isolates and characterization of the PHA produced by strains positive on the polymerase chain reaction (PCR) was envisaged . METHODS AND RESULTS: Different bacteria isolated from soil were screened by PCR using two sets of primers designed for Bacillus megaterium . Amongst 23 isolates examined, the DNA of 12 isolates reacted positively with the primers giving amplicons identical in size to that obtained from B . megaterium . The isolates which were identified as strains of B . sphaericus, B . circulans, B . brevis and B . licheniformis, produced 11- 41% of PHA in biomass, in sucrose-containing medium, over a growth period of 24-72 h . The nature of the PHA thus produced was analyzed by Fourier transform infrared spectroscopy, gas chromatography and by nuclear magnetic resonance (NMR) and found to contain polyhydroxy butyrate and polyhydroxyvalerate . CONCLUSIONS: The results indicate that most of our isolates from different species contained the B . megaterium type of PHA synthase . Bacillus licheniformis appeared to belong to another group as it did not react with both sets of primers . SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the universality of the B . megaterium type of PHA synthase in soil isolates of Bacillus . Some variations were also found.

Acta Microbiol Pol, 2002, 51(3), 275 - 83
Antifungal activity of Bacillus coagulans against Fusarium sp; Czaczyk K et al.; The antifungal activity of Bacillus coagulans against three pathogenic species of Fusarium was examined . Fungal growth was determined by colony forming units, dry matter and ergosterol level . Biosynthesis of Fusarium mycotoxins was also investigated . The strongest inhibition of fungal growth was noticed when Bacillus coagulans was co-inoculated at the beginning of culture . Estimation of ergosterol level as a determinant of fungal growth showed the greatest degree of Fusarium sp . inhibition . Addition of Bacillus coagulans to Fusarium culmorum culture inhibits the DON (deoxynivalenol) production.

Scand J Infect Dis, 2002, 34(12), 925 - 7
Stenotrophomonas maltophilia endocarditis; Crum NF et al.; Stenotrophomonas maltophilia is a gram-negative bacillus that is increasingly associated with serious nosocomial infections, especially in immunocompromised patients; however, the occurrence of endocarditis due to this organism is rare . This report describes a case of S . maltophilia endocarditis associated with a central venous catheter . The literature on Stenotrophomonas endocarditis is reviewed . Given the high morbidity and mortality of these infections, early antibiotic therapy utilizing trimethoprim-sulfamethoxazole, along with a second agent and removal of prosthetic devices, is recommended.

FEMS Microbiol Rev, 2003 Jan, 26(5), 419 - 32
PCR-based identification of Bacillus thuringiensis pesticidal crystal genes; Porcar M et al.; The polymerase chain reaction (PCR) is a molecular tool widely used to characterize the insecticidal bacterium Bacillus thuringiensis . This technique can be used to amplify specific DNA fragments and thus to determine the presence or absence of a target gene . The identification of B . thuringiensis toxin genes by PCR can partially predict the insecticidal activity of a given strain . PCR has proven to be a rapid and reliable method and it has largely substituted bioassays in preliminary classification of B . thuringiensis collections . In this work, we compare the largest B . thuringiensis PCR-based screenings, and we review the natural occurrence of cry genes among native strains . We also discuss the use of PCR for the identification of novel cry genes, as well as the potential of novel technologies for the characterization of B . thuringiensis strains.

Anal Chem, 2003 Feb 1, 75(3), 694 - 7
Use of bioactive glass slides for matrix-assisted laser desorption/ionization analysis: application to microorganisms; Afonso C et al.; Glass slides are widely used in high-throughput analysis and are available commercially with surfaces activated, etched, and channeled . Thin glass microscope slides are shown here to be suitable sample supports for matrix-assisted laser desorption/ionization (MALDI) mass spectrometry . As a demonstration, lectins immobilized on glass slides with activated surfaces are used to concentrate and purify agglutinated Bacillis spores . It is expected that such slides will provide a rapid, inexpensive way to evaluate and implement new strategies involving MALDI MS readout.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 2002 Feb, 22(2), 126 - 8
{Experimental study on liver protective effect of Artemisia rupestris extract}; Israpil et al.; OBJECTIVE: To explore the protective effect of Artemisia rupestris extract (ARE) on chemical liver injury and immunological hepatitis . METHODS: Mice model of immunological hepatitis was established by lipopolysaccharide plus Bacillus Calmette-Guerin vaccine (BCG), and the models of liver injury was induced by D-galactosamine or carbon tetrachloride (CCl4) in mice and rats . The effects of ARE in preventing and treating immunological hepatitis and protecting liver injury were observed . RESULTS: ARE significantly lowered the serum alanine transaminase (ALT) in immunological hepatitis (P < 0.05), CCl4 induced liver injury model mice and rats (P < 0.01), and in D-galactosamine induced liver injury mice (P < 0.05) . CONCLUSION: ARE has significantly protective effect against chemical liver injury and could treat immunological hepatitis, but with no effect in preventing immunological hepatitis . These might be the partial pharmacologic basis for using ARE in treating hepatitis.

Srp Arh Celok Lek, 2002 Jul-Aug, 130(7-8), 265 - 9
{Tuberculous lymphadenitis of the cervical mediastinum.}; Ivanovic-Krstic B et al.; Tuberculous lymphadenitis is an uncommon form of extrapulmonary tuberculosis . We report on a case of isolated Cervicomediastinal tuberculous lymphadenitis without parenhimal involvement . A 49-year old woman was hospitalized with a four week history of night sweats and weight loss . Plain chest radiography disclosed mediastinal mass of the right side . Echocardiographic examination revealed a soft tissue mass below the ascending aorta and aortic arch causing moderate narrowing of the right ventricular outflow tract and the main pulmonary artery . Computed tomographic scanning showed soft tissue mass in the middle mediastinum surrounding the great vessels extending posteriorely with moderate compression of trachea . Histological examination of the cervical mass revealed caseating granuloma containing acid-alcohol fast bacillus . Antituberculous chemotherapy was started.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 2002 Apr, 22(4), 286 - 8
{Study of protective effect and mechanism for baicalin on bacillus pertussis infected brain tissue and its dose-effect relationship}; Yin F et al.; OBJECTIVE: To study the possible mechanism of protective effect for Baicalin on Bacillus pertussis (BP) infected brain tissue and the dose-effect relationship . METHODS: Brain tissues slices were divided into 7 groups: (1) the normal group; (2) the model group: infected by 10% BP; (3) the baicalin group, which was pretreated with baicalin, infected by BP and subdivided into 5 sub-groups according to different doses of baicalin used; (4) the glutamic acid group: cultured with glutamic acid; (5) the baicalin plus glutamic acid group; (6) the peroxide group: cultured with hydrogen peroxide; and (7) the baicalin plus peroxide group . The lactate dehydrogenase (LDH) content in the supernatant of culture was determined and quantitative protein determination was conducted . RESULTS: The LDH releasing was higher in the model group, glutamic acid group and peroxide group as compared with that in the normal group, 15.10 +/- 4.89 u/g . protein (the same unit below), 15.49 +/- 5.66 and 16.54 +/- 5.47 vs 6.10 +/- 2.87 respectively (P < 0.01) . After being pretreated with 0.25 mmol/L baicalin, LDH level decreased significantly to 8.65 +/- 2.43, which was significantly different from that in the model group (P < 0.01), LDH was also decreased in the baicalin plus glutamic acid group (9.93 +/- 2.89) and baicalin plus peroxide group (9.54 +/- 2.82), which was significantly lower than that in the glutamic acid group and the peroxide group respectively (P < 0.01) . CONCLUSION: Pretreatment of baicalin has protective effect on BP caused nerve cell injury in rat brain slices, the protection is possibly related with the reduction of glutamic acid and hydrogen peroxide induced damage on nerve cells in vitro.

Proc Natl Acad Sci U S A, 2003 Mar 18, 100(6), 3461 - 6 Epub 2003 Feb 12.
Kinetics of size changes of individual Bacillus thuringiensis spores in response to changes in relative humidity; Westphal AJ et al.; Using an automated scanning microscope, we report the surprising result that individual dormant spores of Bacillus thuringiensis grow and shrink in response to increasing and decreasing relative humidity . We simultaneously monitored the size of inorganic calibration particles . We found that the spores consistently swell in response to increased relative humidity, and shrink to near their original size on reexposure to dry air . Although the dispersion of swelling amplitudes within an ensemble of spores is wide (approximately 30% of the average amplitude), amplitudes for individual spores are highly correlated between different swelling episodes, suggesting that individual spores respond consistently to changes in humidity . We find evidence for two distinct time scales for swelling: one with a time scale of no more than approximately 50 s, and another with a time scale of approximately 8 min . We speculate that these two mechanisms may be due to rapid diffusion of water into the spore coat + cortex, followed by slower diffusion of water into the spore core, respectively . Humidity-dependent swelling may account for the greater kill effectiveness of spores by gas-phase chlorine dioxide, formaldehyde, and ethylene oxide at very high relative humidity.

FEMS Microbiol Lett, 2003 Jan 21, 218(1), 161 - 5
Microorganisms cultured from stratospheric air samples obtained at 41 km; Wainwright M et al.; Samples of air removed from the stratosphere, at an altitude of 41 km, were previously found to contain viable, but non-cultureable bacteria (cocci and rods) . Here, we describe experiments aimed at growing these, together with any other organisms, present in these samples . Two bacteria (Bacillus simplex and Staphylococcus pasteuri) and a single fungus, Engyodontium album (Limber) de Hoog were isolated from the samples . Although the possibility of contamination can never be ruled out when space-derived samples are studied on earth, we are confident that the organisms originated from the stratosphere . Possible mechanisms by which these organisms could have attained such a height are discussed.

FEMS Microbiol Lett, 2003 Jan 21, 218(1), 39 - 45
Conversion of the nitrogen content in liquid manure into biomass and polyglutamic acid by a newly isolated strain of Bacillus licheniformis; Hoppensack A et al.; Extensive spreading of liquid manure onto agricultural fields causes eutrophication of ground and surface water and also pollution of the atmosphere due to the high ammonium nitrogen content . A poly(gamma-glutamic acid) (PGA)-producing strain of Bacillus licheniformis was isolated in this study and investigated for its ability to reduce the ammonium nitrogen by converting ammonium into biomass and PGA as depot forms of nitrogen . In batch cultivations swine manure and an optimized mineral salts medium were used for PGA production . For example the cultivation of B . licheniformis strain S2 in liquid manure, which was modified by adding of 18 g citrate and 80 g glycerol l(-1) and exhibited a carbon to nitrogen ratio of 15.5:1, led to severe reduction of the ammonium content from 2.83 to 0.1 g x l(-1) and to the production of 0.16 g PGA and 7.5 g cell dry mass l(-1) within 410 h . Approximately 28% (w/w) of the total nitrogen was converted into cellular biomass, whereas 0.1% (w/w) was used for the production of PGA . In addition, approximately 33% (w/v) of the original ammonium was lost by stripping.

Theor Appl Genet, 2002 Mar, 104(4), 727 - 734
Inheritance and expression of the cry1Ab gene in Bt ( Bacillus thuringiensis) transgenic rice; Wu G et al.; The inheritance and expression patterns of the cry1Ab gene were studied in the progenies derived from different Bt ( Bacillus thuringiensis) transgenic japonica rice lines under field conditions . Both Mendelian and distorted segregation ratios were observed in some selfed and crossed F(2) populations . Crosses between japonica intra-subspecies had no significant effect on the segregation ratios of the cry1Ab gene, but crossing between japonica and indicainter-subspecies led to distorted segregation of the cry1Ab gene in the F(2)population . Field-release experiments indicated that the cry1Ab gene was stably transmitted in an intact manner via successive sexual generations, and the concentration of the Cry1Ab protein was kept quantitatively stable up to the R(6)generation . The cry1Ab gene, driven by the maize ubiquitinpromoter, displayed certain kinds of spatial and temporal expression patterns under field conditions . The content of the Cry1Ab protein varied in different tissues of the main stems, the primary tillers and the secondary tillers . Higher levels of the Cry1Ab protein were found in the stems, leaves and leaf sheaths than in the roots, while the lowest level was detected in grains at the maturation stage . The content of the Cry1Ab protein in the leaves peaked at the booting stage and was lowest at the heading stage . Furthermore, the Cry1Ab content of cry1Ab expression in different tissues of transgenic rice varied individually with temperature.

Theor Appl Genet, 2002 Aug, 105(2-3), 258 - 264 Epub 2002 Jun 14.
Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1C proteins; Cao J et al.; This study was undertaken to determine the effects of pyramiding two Bacillus thuringiensis (Bt) genes in the same plant on the production of Bt proteins and the control of diamondback moths (DBM, Plutella xylostella) resistant to one or the other protein . Broccoli lines carrying both cry1Ac and cry1C Bt genes were produced by sexual crosses of cry1Ac- and cry1C-transgenic plants . Plants containing both genes were selected by tests for resistance to kanamycin and hygromycin, and confirmed by PCR analysis for the Bt genes . Both cry1Ac and cry1C mRNAs were detected in the hybrid lines, and Cry1Ac and Cry1C proteins were stably produced at levels comparable to the parental plants . Plants producing both Cry1Ac and Cry1C proteins caused rapid and complete mortality of DBM larvae resistant to Cry1A or Cry1C, and suffered little or no leaf damage . These plants, in combination with the resistant DBM populations available, will allow greenhouse or field studies of resistance management strategies involving gene pyramiding.

J Invertebr Pathol, 2003 Jan, 82(1), 63 - 71
Characterization of cry1, cry2, and cry9 genes in Bacillus thuringiensis isolates from China; Wang J et al.; Bacillus thuringiensis isolates from different ecological regions and sources of China were analyzed to study the distribution and diversity of cry genes and to detect the presence of novel cry genes . Strains containing cry1-type genes were the most abundant and represent 237 of the 310 B . thuringiensis isolates (76.5%) . About 70 and 15.5% of the isolates contained a cry2 gene or cry9 gene, respectively, while 10.0% of the strains did not contain a cry1, cry2, or cry9 gene . Among the cry1 containing isolates, cry1A (67.7%), cry1I (60.6%), cry1C (43.9%), and cry1D (39.4%) genes were the most abundant . Forty-three different cry1 gene profiles were detected in this collection . Several cry1 genes were associated at a high frequency, such as the cry1C-cry1D and cry1A-cry1I gene combination . The cry1A and cry2 amplicons were digested with selected restriction enzymes to examine sequence diversity . Based on this RFLP analysis, one novel cry1A-type gene was observed.

J Invertebr Pathol, 2003 Jan, 82(1), 57 - 62
Correlation between serovars of Bacillus thuringiensis and type I beta-exotoxin production; Hernandez CS et al.; beta-Exotoxin is a thermostable metabolite produced by some strains of Bacillus thuringiensis . Because of vertebrate toxicity, most commercial preparations of B . thuringiensis are prepared from isolates that do not produce beta-exotoxin . The aim of the present study was to find out the possible relationship between serovars of B . thuringiensis and beta-exotoxin production . A specific HPLC assay for type I beta-exotoxin has been used to detect this exotoxin in supernatants from final whole cultures of 100 strains belonging to four serovars of B . thuringiensis: thuringiensis, kurstaki, aizawai, and morrisoni . For each serovar, 25 strains randomly chosen from two Spanish collections were analyzed . Frequency of beta-exotoxin production was higher in B . thuringiensis serovar thuringiensis, whereas only two strains from serovar kurstaki showed beta-exotoxin production . None of the 25 strains belonging to serovars aizawai and morrisoni was found to produce this compound . Along with data from other studies, serovars can be classified as "common," "seldom," or "rare" beta-exotoxin producers . The serovar-dependent beta-exotoxin production is discussed in relation to the evolutionary process of serovar differentiation, the plasmid compatibility and limited plasmid exchange between serovars, and with the serovar-dependent regulation of plasmid-encoded genes.

J Invertebr Pathol, 2003 Jan, 82(1), 1 - 11
Characterization and comparison of midgut proteases of Bacillus thuringiensis susceptible and resistant diamondback moth (Plutellidae: Lepidoptera); Mohan M et al.; The midgut proteases of the Bacillus thuringiensis resistant and susceptible populations of the diamondback moth, Plutella xylostella L . were characterized by using protease specific substrates and inhibitors . The midgut contained trypsin-like proteases of molecular weights of 97, 32, 29.5, 27.5, and 25 kDa . Of these five proteases, 29.5 kDa trypsin-like protease was the most predominant in activation of protoxins of Cry1Aa and Cry1Ab . The activation of Cry1Ab protoxin by midgut protease was fast (T(1/2) of 23-24 min) even at a protoxin:protease ratio of 250:1 . The protoxin activation appeared to be multi-step process, and at least seven intermediates were observed before formation of a stable toxin of about 57.4 kDa from protoxin of about 133 kDa . Activation of Cry1Aa was faster than that of Cry1Ab on incubation of protoxins with midgut proteases and bovine trypsin . The protoxin and toxin forms of Cry proteins did not differ in toxicity towards larvae of P . xylostella . The differences in susceptibility of two populations to B . thuringiensis Cry1Ab were not due to midgut proteolytic activity . Further, the proteolytic patterns of Cry1A protoxins were similar in the resistant as well as susceptible populations of P . xylostella.

Adv Space Res, 2003, 31(1), 235 - 40
High temperature effect on microflora of radish root-inhabited zone and nutrient solutions for radish growth; Borodina EV et al.; The effect of high temperatures (35 and 45 degrees C) on microflora of the root zone of radish plants grown in phytotron was evaluated by the response of microorganisms from 9 indicator groups . Phytotron air temperature elevated to 35 degrees C for 20 hours caused no significant changes in qualitative and quantitative composition of the root microflora in experimental plants . By the end of the experiment, the species diversity of microflora had changed . The amount of phytopathogenic microorganisms decreased which can be interpreted as more stable co-existence of microflora with plants . The numbers of microbes from other indicator groups was in dynamic equilibrium . The plants' condition did not deteriorate either . Exposure to the temperature of 45 degrees C for 7 hours have been found to change the numbers and species diversity in the radish root zone microflora . The microorganisms were observed to increase their total numbers at the expense of certain indicator groups . Bacteria increased spore forms at the stage of spores . Colon bacillus bacteria of increased their numbers by the end of experiment by an order . By the end of experiment the roots of experiment plants had microscopic fungi from Mucor, Aspergillus, Trichoderma, Cladosporium genera . The observed changes in the microbial complex seem to be associated with the changes of root emissions and general deterioration of the plants' condition . It is suggested that the response of the microorganisms can be indicative of the condition of plants under investigation . c2002 Published by Elsevier Science Ltd on behalf of COSPAR.

Yao Xue Xue Bao, 2001 Feb, 36(2), 81 - 3
{Effect of stilbene polymer (Gn-3) on experimental liver injuries in mice}; Hou YN et al.; AIM: To study the protective effect of Gn-3 (a stilbene polymer isolated from Gnetum parvifolium) against liver injury induced by CCl4, N-acetyl-p-aminophenol (APAP) and Bacillus Calmette-Guerin (BCG) plus bacterial lipopolysaccharide (LPS) in mice . METHODS: The experimental model of liver injury were induced by 0.1% CCl4 i.p . (10 mL.kg-1.d-1 for 3d), APAP i.p . (150 mg.kg-1) or BCG (5 mg) plus LPS (7.5 micrograms) in mice . The levels of ALT in serum, MDA and GSH in liver tissues were detected . The histopathologic changes were observed by light microscope . RESULTS: Gn-3 was shown to markedly reduce the elevated serum ALT levels, liver tissue MDA and improve the histopathological changes in all the three experimental liver injury models . No effect of Gn-3 was observed on the liver GSH level in liver injury mice . CONCLUSION: Gn-3 was found to inhibit the development of liver injury caused by CCl4, APAP, or BCG plus LPS . This means that Gn-3 has liver protective effects.

Biochemistry, 2003 Feb 18, 42(6), 1603 - 10
Altering substrate specificity of phosphatidylcholine-preferring phospholipase C of Bacillus cereus by random mutagenesis of the headgroup binding site; Antikainen NM et al.; PLC(Bc) is a 28.5 kDa monomeric enzyme that catalyzes the hydrolysis of the phosphodiester bond of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine to provide a diacylglycerol and the corresponding phosphorylated headgroup . Because single replacements of Glu4, Tyr56, and Phe66 in the headgroup binding pocket led to changes in substrate specificity {Martin et al . (2000) Biochemistry 39, 3410-3415}, a combinatorial library of approximately 6000 maltose binding protein-PLC(Bc) fusion protein mutants containing random permutations of these three residues was generated to identify PLC(Bc) mutants with altered specificity profiles and high catalytic activities . Members of this library were screened for hydrolytic activity toward the water soluble substrates C6PC, C6PE, and C6PS using a novel protocol that was conducted in a 96-well format and featured the in situ cleavage of the fusion protein to release the mutant PLC(Bc)s . Ten mutant enzymes that exhibited significant preferences toward C6PE or C6PS were selected and analyzed by steady-state kinetics to determine their specificity constants, k(cat)/K(M) . The C6PS selective clones E4G, E4Q/Y56T/F66Y, and E4K/Y56V exhibited higher specificity constants toward C6PS than wt, whereas Y56T, F66Y, and Y56T/F66Y were C6PE selective and had comparable or higher specificity constants than wt for C6PE . The corresponding wt residues were singly reinserted back into the E4Q/Y56T/F66Y and E4K/Y56V mutants via site-directed mutagenesis, and the E4Q/F66Y mutant thus obtained exhibited a 10-fold higher specificity constant toward C6PS than wt, a value significantly higher than other PLC(Bc) mutants . On the basis of available data, an aromatic residue at position 66 appears important for significant catalytic activity toward all three substrates, especially C6PC and C6PE . The charge of residue 4 also appears to be a determinant of enzyme specificity as a negatively charged residue at this position endows the enzyme with C6PC and C6PE preference, whereas a polar neutral or positively charged residue results in C6PS selectivity . Replacing Tyr56 with Val, Ala, Thr, or Ser greatly reduces activity toward C6PC . Thus, the substrate specificity of PLC(Bc) can be modulated by varying three of the amino acid residues that constitute the headgroup binding pocket, and it is now apparent that this enzyme is not evolutionarily optimized to hydrolyze phospholipids with ethanolamine or serine headgroups.

Otolaryngol Pol, 2002, 56(6), 745 - 8
{The achievements of eminent Warsaw physicians in otolaryngology at the turn of the 19th and 20th century}; Kierzek A; The scientific achievements of many Warsaw otolaryngologists in the second part of the 19th and in the beginning of the 20th century are presented . Karol Benni's (1843-1916) contribution to the world otology is described . The famous Warsaw laryngologist Teodor Heryng (1847-1925) was a creator of a new operating method of the larynx tuberculosis by curettage . He was also a propagator of the diaphanoscopy (transillumination) of maxillary sinuses . Ludwik Guranowski (1853-1926) . a distinguished otiatrist performed a first myringoplasty in the Polish territories and explained the causes of external otitis by bacillus pyocyaneous . Jan Sedziak's (1861-1932) work on malignant neoplasms of larynx was given a prize at international congress in Tuluse in 1893 . Zdzislaw Dmochowski (1864-1924) described the pathologic anatomy changes in maxillary sinuses . Benni and Alfred Sokolowski (1849-1924) were the authors of several chapters in foreign medical textbooks . Heryng's inhalation apparatuses exhibited in Paris are described . Sedziak, Przemyslaw Pieniazek (1850-1916) and Teodor Heiman (1848-1917) were also the authors of articles in foreign otolaryngological journals written in connection with the jubilee of famous otolaryngologists: Leopold von Schroetter, Bernhard Fraenkel and Hermann Schwartze . The problem of focal infection in otolaryngology described by Feliks Erbrich (1874-1938) is briefly pointed out.

Res Microbiol, 2003 Jan-Feb, 154(1), 49 - 53
Nutritional and cultural parameters influencing antidipteran delta-endotoxin production; Ozkan M et al.; In this study, various nutritional and cultural parameters influencing diptera-specific delta-endotoxin synthesis by Bacillus thuringiensis subsp . israelensis HD500 were investigated . Of various inorganic nitrogen sources, the highest yields of Cry11Aa and Cry4Ba proteins were obtained on (NH(4))(2)HPO(4) . Among carbon sources, inulin, dextrin, maltose, lactose, sucrose, whey and glycerol were all stimulatory, while glucose, starch and molasses were suppressive . High concentrations of inorganic phosphate (50 to 100 mM K(2)HPO(4)) were required for an effective synthesis of Cry4Ba . Mn was the most critical element for the biosynthesis of both toxins at 10(-6) M concentration . Mg and Ca favored production when provided at 8 x 10(-3) M and 5.5 x 10(-4) M concentrations, respectively, while Fe, Zn and Cu negatively influenced biosynthesis . Cry4-toxin synthesis was best at neutral pH and also when the organism was grown at 25 degrees C . Throughout the study, the extent of growth and sporulation of the producer organism was also monitored.

Hunan Yi Ke Da Xue Xue Bao, 2002 Feb 28, 27(1), 9 - 12
{Molecular conformation and expression efficiency in the hepatocyte targeting gene drug}; Lu FG et al.; OBJECTIVE: To modulate the molecular conformation of the hepatocyte targeting drug to increase exogenous gene expression efficiency in the targeted cell . METHODS: Some specific auxiliary molecules were added to the gene drug its molecular liquid conformation was investigated under the electron-microscope and they were transfected into human hepatoma cell line BEL-7402 in vitro . We detected IFN-gamma gene expression product by ELISA and screened out the most efficient drug molecules . RESULTS: Due to various concentrations of auxiliary molecules, the conformation of hepatocyte targeting drug changed, including floccule, globulous, stringbeads, bacillform and chromosome-like mixture . While the most efficient expression conformation was the bacilliform and chromosome-like mixture and didn't need to inhibit the lysozyme activity, the expression efficiency of such drug molecules was much higher than that of the liposome targeting vector . CONCLUSION: The molecular drug conformation had much influence on targeted gene expression efficiency in gene therapy . Bacilliform and chromosome-like mixture may be the most efficient expression conformation to construct drug molecules in ASOR targeted hepatocyte gene therapy . So we put forward a new concept "chromosome mimic conformation" in screening out the most efficient molecular drug conformation in gene therapy.

Science, 2003 Feb 7, 299(5608), 900 - 2
Yield effects of genetically modified crops in developing countries; Qaim M et al.; Onfarm field trials carried out with Bacillus thuringiensis (Bt) cotton in different states of India show that the technology substantially reduces pest damage and increases yields . The yield gains are much higher than what has been reported for other countries where genetically modified crops were used mostly to replace and enhance chemical pest control . In many developing countries, small-scale farmers especially suffer big pest-related yield losses because of technical and economic constraints . Pest-resistant genetically modified crops can contribute to increased yields and agricultural growth in those situations, as the case of Bt cotton in India demonstrates.

Am J Respir Crit Care Med, 2003 May 15, 167(10), 1355 - 9 Epub 2003 Feb 05.
High plasma osteopontin level and its relationship with interleukin-12-mediated type 1 T helper cell response in tuberculosis; Koguchi Y et al.; Osteopontin (OPN, also known as Eta-1), a noncollagenous matrix protein produced by macrophages and T lymphocytes, is expressed in granulomatous lesions caused by Mycobacterium tuberculosis infection . In the present study, we compared plasma concentrations of OPN in patients with active pulmonary tuberculosis with those of healthy control subjects and patients with sarcoidosis, another disease associated with granuloma formation . Plasma OPN levels were significantly higher in patients with tuberculosis (n = 48) than in control subjects (n = 34) and patients with sarcoidosis (n = 20) . OPN levels correlated well with severity of pulmonary tuberculosis, as indicated by the size of lung lesions on chest X-ray films . Furthermore, chemotherapy resulted in a significant fall in plasma OPN levels . In patients with tuberculosis, plasma OPN concentrations correlated significantly with those of interleukin (IL)-12 . In vitro experiments showed that OPN production by peripheral blood mononuclear cells infected with Mycobacterium bovis bacillus Calmette-Guerin preceded the synthesis of IL-12 and interferon-gamma and that the neutralizing anti-OPN monoclonal antibody significantly reduced the production of IL-12 and interferon-gamma . Our results suggest that OPN may be involved in the pathologic process associated with active pulmonary tuberculosis by inducing IL-12-mediated type 1 T helper cell responses.

J Colloid Interface Sci, 2002 Dec 15, 256(2), 237 - 48
Surface chemical studies on sphalerite and galena using extracellular polysaccharides isolated from Bacillus polymyxa; Santhiya D et al.; Adsorption, electrokinetic, microflotation, and flocculation studies have been carried out on sphalerite and galena minerals using extracellular polysaccharides (ECP) isolated from Bacillus polymyxa . The adsorption density of ECP onto galena is found to be higher than that onto sphalerite . The adsorption of ECP onto sphalerite is found to increase from pH 3 to about pH 7, where a maximum is attained, and thereafter continuously decreases . With respect to galena, the adsorption density of ECP steadily increases with increased pH . The addition of ECP correspondingly reduces the negative electrophoretic mobilities of sphalerite and galena in absolute magnitude without shifting their isoelectric points . However, the magnitude of the reduction in the electrophoretic mobility values is found to be greater for galena compared to that for sphalerite . Microflotation tests show that galena is depressed while sphalerite is floated using ECP in the entire pH range investigated . Selective flotation tests on a synthetic mixture of galena and sphalerite corroborate that sphalerite could be floated from galena at pH 9-9.5 using ECP as a depressant for galena . Flocculation tests reveal that in the pH range 9-11, sphalerite is dispersed and galena is flocculated in the presence of ECP . Dissolution tests indicate release of the lattice metal ions from galena and sphalerite, while co-precipitation tests confirm chemical interaction between lead or zinc ions and ECP . Fourier transform infrared spectroscopic studies provide evidence in support of hydrogen bonding and chemical interaction for the adsorption of ECP onto galena/sphalerite surfaces.

Bioresour Technol, 2003 May, 88(1), 9 - 15
Xylan-hydrolyzing enzyme system from Bacillus pumilus CBMAI 0008 and its effects on Eucalyptus grandis kraft pulp for pulp bleaching improvement; Duarte MC et al.; The extracellular productions of beta-xylanase, beta-xylosidase, beta-glucosidase, beta-mannanase, arabinosidase, alpha-glucuronidase, alpha-galactosidase and Fpase from Bacillus pumilus CBMAI 0008 were investigated with three different xylan sources as substrate . The enzymatic profiles on birchwood, Eucalyptus grandis and oat were studied at alkaline and acidic pH conditions . B . pumilus CBMAI 0008 grown on the three carbon sources produced mainly beta-xylanase . At pH 10, the levels of xylanase were 328, 160 and 136 U/ml, for birch, oat and E . grandis, respectively . beta-Mannanase production was induced on E . grandis (5 U/ml) and arabinofuranosidase on oat (5 U/ml) . Although small quantities of alpha-glucuronidase had been produced at pH 10, activity at pH 4.8 was 1.5 U/ml, higher than observed for Aspergillus sp . in literature reports . Preliminary assays carried out on E . grandis kraft pulp from an industrial paper mill (RIPASA S.A . Celulose e Papel, Limeira, SP, Brazil) showed a reduction of 0.3% of chlorine use in the pulp treated with the enzymes, resulting in increased brightness, compared to conventional bleaching . The enzymes were more efficient if applied before the initial bleaching sequence, in a non-pre-oxygenated pulp.

Proc Natl Acad Sci U S A, 2003 Feb 18, 100(4), 1519 - 23 Epub 2003 Feb 05.
Long-term regional suppression of pink bollworm by Bacillus thuringiensis cotton; Carriere Y et al.; Despite the potentially profound impact of genetically modified crops on agriculture and the environment, we know little about their long-term effects . Transgenic crops that produce toxins from Bacillus thuringiensis (Bt) to control insects are grown widely, but rapid evolution of resistance by pests could nullify their benefits . Here, we present theoretical analyses showing that long-term suppression of pest populations is governed by interactions among reproductive rate, dispersal propensity, and regional abundance of a Bt crop . Supporting this theory, a 10-year study in 15 regions across Arizona shows that Bt cotton suppressed a major pest, pink bollworm (Pectinophora gossypiella), independent of demographic effects of weather and variation among regions . Pink bollworm population density declined only in regions where Bt cotton was abundant . Such long-term suppression has not been observed with insecticide sprays, showing that transgenic crops open new avenues for pest control . The debate about putative benefits of Bt crops has focused primarily on short-term decreases in insecticide use . The present findings suggest that long-term regional pest suppression after deployment of Bt crops may also contribute to reducing the need for insecticide sprays.

Appl Environ Microbiol, 2003 Feb, 69(2), 1331 - 4
Recombinant strain of Bacillus thuringiensis producing Cyt1A, Cry11B, and the Bacillus sphaericus binary toxin; Park HW et al.; A novel recombinant Bacillus thuringiensis subsp . israelensis strain that produces the B . sphaericus binary toxin, Cyt1Aa, and Cry11Ba is described . The toxicity of this strain (50% lethal concentration {LC(50)} = 1.7 ng/ml) against fourth-instar Culex quinquefasciatus was higher than that of B . thuringiensis subsp . israelensis IPS-82 (LC(50) = 7.9 ng/ml) or B . sphaericus 2362 (LC(50) = 12.6 ng/ml).

Appl Environ Microbiol, 2003 Feb, 69(2), 1197 - 205
Use of a promoter trap to identify Bacillus cereus genes regulated by tomato seed exudate and a rhizosphere resident, Pseudomonas aureofaciens; Dunn AK et al.; The goal of this study was to identify genes in Bacillus cereus, a bacterium commonly associated with plant seeds and roots, that are affected by compounds originating from a host plant, tomato, or another rhizosphere resident, Pseudomonas aureofaciens . We constructed a B . cereus chromosomal DNA library in a promoter-trap plasmid, pAD123, which contains a promoterless version of the green fluorescent protein (GFP) gene, gfpmut3a . The library was screened by using fluorescence-activated cell sorting for clones showing a change in GFP expression in response to either tomato seed exudate or culture supernatant of P . aureofaciens strain 30-84 . We identified two clones carrying genes that were induced by the presence of tomato seed exudate and nine clones carrying genes that were repressed by P . aureofaciens culture supernatant . A clone chosen for further study contained an open reading frame, designated lipA, that encodes a deduced protein with a lipoprotein signal peptide sequence similar to lipoproteins in B . subtilis . Expression of gusA under control of the lipA promoter increased twofold when cells were exposed to tomato seed exudate and in a concentration-dependent manner when exposed to a mixture of amino acids . When the wild type and a 10-fold excess of a lipA mutant were applied together to tomato seeds, 2 days after planting, the wild type displayed medium-dependent culturability, whereas the lipA mutant was unaffected . This study demonstrates the power of a promoter trap to identify genes in a gram-positive bacterium that are regulated by the biotic environment and resulted in the discovery of lipA, a plant-regulated gene in B . cereus.

Appl Environ Microbiol, 2003 Feb, 69(2), 1100 - 7
Small, acid-soluble proteins as biomarkers in mass spectrometry analysis of Bacillus spores; Hathout Y et al.; The use of 1 N HCl for extraction of small, acid-soluble proteins (SASP) from different Bacillus spore species was examined . The extracts were analyzed by high-performance liquid chromatography and matrix-assisted laser desorption mass spectrometry and were found to be both qualitatively and quantitatively superior to extraction by acetonitrile-5% trifluoroacetic acid (70:30, vol/vol) . Both major and minor alpha/beta- and gamma-type SASP were characterized by their molecular masses or tryptic peptide maps and by searches of both protein and unannotated genome databases . For all but 1 pair (B . cereus T and B . thuringiensis subsp . Kurstaki) among the 11 variants studied the suites of SASP masses are distinctive, consistent with the use of these proteins as potential biomarkers for spore identification by mass spectrometry.

Appl Environ Microbiol, 2003 Feb, 69(2), 1023 - 9
Cloning, sequencing, and expression of the chitinase gene chiA74 from Bacillus thuringiensis; Barboza-Corona JE et al.; The endochitinase gene chiA74 from Bacillus thuringiensis serovar kenyae strain LBIT-82 was cloned in Escherichia coli DH5 alpha F' . A sequence of 676 amino acids was deduced when the gene was completely sequenced . A molecular mass of 74 kDa was estimated for the preprotein, which includes a putative 4-kDa signal sequence located at the N terminus . The deduced amino acid sequence showed high degree of identity with other chitinases such as ChiB from Bacillus cereus (98%) and ChiA71 from Bacillus thuringiensis serovar pakistani (70%) . Additionally, ChiA74 showed a modular structure comprised of three domains: a catalytic domain, a fibronectin-like domain, and a chitin-binding domain . All three domains showed conserved sequences when compared to other bacterial chitinase sequences . A ca . 70-kDa mature protein expressed by the cloned gene was detected in zymograms, comigrating with a chitinase produced by the LBIT-82 wild-type strain . ChiA74 is active within a wide pH range (4 to 9), although a bimodal activity was shown at pH 4.79 and 6.34 . The optimal temperature was estimated at 57.2 degrees C when tested at pH 6 . The potential use of ChiA74 as a synergistic agent, along with the B . thuringiensis insecticidal Cry proteins, is discussed.

J Biol Chem, 2003 Apr 11, 278(15), 12968 - 76 Epub 2003 Feb 04.
Crystal structure of two ternary complexes of phosphorylating glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus with NAD and D-glyceraldehyde 3-phosphate; Didierjean C et al.; The crystal structure of the phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus was solved in complex with its cofactor, NAD, and its physiological substrate, D-glyceraldehyde 3-phosphate (D-G3P) . To isolate a stable ternary complex, the nucleophilic residue of the active site, Cys(149), was substituted with alanine or serine . The C149A and C149S GAPDH ternary complexes were obtained by soaking the crystals of the corresponding binary complexes (enzyme.NAD) in a solution containing G3P . The structures of the two binary and the two ternary complexes are presented . The D-G3P adopts the same conformation in the two ternary complexes . It is bound in a non-covalent way, in the free aldehyde form, its C-3 phosphate group being positioned in the P(s) site and not in the P(i) site . Its C-1 carbonyl oxygen points toward the essential His(176), which supports the role proposed for this residue along the two steps of the catalytic pathway . Arguments are provided that the structures reported here are representative of a productive enzyme.NAD.D-G3P complex in the ground state (Michaelis complex).

Biochem J, 2003 May 1, 371(Pt 3), 997 - 1003
Pre-steady-state kinetics of Bacillus licheniformis 1,3-1,4-beta-glucanase: evidence for a regulatory binding site; Abel M et al.; In a previous paper, we reported the first stopped-flow experiments on a Bacillus licheniformis 1,3-1,4-beta-glucanase {Abel, Planas and Christensen (2001) Biochem . J . 357, 195-202} . It was shown that the pre-steady-state kinetics of the 1,3-1,4-beta-glucanase using the substrate 4-methylumbelliferyl 3-O-beta-cellobiosyl-beta-D-glucoside may be explained by a reaction scheme involving an induced fit and the binding of two substrates as well as a second enzymic conformational change, whereas the results definitely could not be explained in terms of the simple double-displacement scheme . In the present study, we report further stopped-flow kinetic results on the glucanase using a series of low-molecular-mass substrates with various leaving groups and varying chain length . The analysis of the resulting data leads to the conclusion that the free enzyme exists in two conformations, one of which binds the substrates rather strongly in a regulatory site, before any productive interactions can take place . This corresponds to an allosteric activation mechanism . With these substrates, however, the productive enzyme-substrate species are also able to change into less active or inactive forms . This may be seen as a feedback inhibitory mechanism.

Curr Microbiol, 2003 Mar, 46(3), 211 - 6
Replacement of methionine 208 in a truncated Bacillus sp . TS-23 alpha-amylase with oxidation-resistant leucine enhances its resistance to hydrogen peroxide; Lin LL et al.; The methionine residues at positions 17, 104, 208, 214, 292, 315, 324, and 446 in the primary amino acid sequence of a truncated Bacillus sp . TS-23 alpha-amylase (His(6)-tagged BLADeltaNC) was changed to oxidative-resistant leucine by site-directed mutagenesis . The mutant enzymes with an apparent molecular mass of approximately 54 kDa were overexpressed in recombinant Escherichia coli . The specific activity for Met315Leu and Met446Leu was decreased by more than 76%, while Met17Leu, Met104Leu, Met208Leu, Met214Leu, Met292Leu, and Met324Leu showed 247, 128, 37, 260, 232, and 241%, respectively, higher activity than the wild-type enzyme . In comparison with wild-type enzyme, a lower K(m) value was observed for all mutant enzymes . The 3.2- and 4.5-fold increases in the catalytic efficiency (k(cat)/K(m)) for Met208Leu and Met324Leu, respectively, were partly contributed by a 68% and 38% decrease in K(m) values . Wild-type enzyme was sensitive to chemical oxidation, but Met208Leu was stable even in the presence of 500 mM H(2)O(2) . Except for Met214Leu, which was quite sensitive to H(2)O(2), the other mutants showed a profile of oxidative inactivation similar to that of the wild-type enzyme . These observations indicate that the oxidative stability of His(6)-tagged BLADeltaNC can be improved by replacement of the critical methionine residue with leucine.

Curr Microbiol, 2003 Mar, 46(3), 169 - 73
Purification and characterization of an extracellular alkaline serine protease with dehairing function from Bacillus pumilus; Huang Q et al.; An extracellular alkaline serine protease (called DHAP), produced by a Bacillus pumilus strain, demonstrates significant dehairing function . This protease is purified by hydrophobic interaction chromatography, ion exchange, and gel filtration . DHAP had a pI of 9.0 and a molecular weight of approximately 32,000 Dalton . It shows maximal activity at pH 10 and with a temperature of 55 degrees C; the enzyme activity can be completely inhibited by phenylmethylsulfonyl fluoride (PMSF) and diisopropyl fluorophosphates (DFP) . The first 20 amino acid residues of the purified DHAP have been determined with a sequence of AQTVPYGIPQIKAPAVHAQG . Alignment of this sequence with other alkaline protease demonstrates its high homology with protease from another B . pumilus strain.

Dan Medicinhist Arbog . 2002;:164-70.
{The human in the art: Henrik Have's painting for a medical thesis}; Kristiansen JE et al.; A painting made by the Danish painter and author Henrik Have (born 1946) illustrates the front page of Jette E . Kristiansen's medical thesis The Antimicrobial Activity of Psychotherapeutic Drugs and Stereo-isomeric Analogues (1990) . The painting illustrates beautifully that art and science can go hand in hand . Even very complicated chemical, pharmacological and microbiological questions can be expressed by means of colours and by means of symbols used in everyday life, such as a spiral (DNA), or a pair of hands expressing working together . Prayers, wishes and the most difficult questions in receptor stereo-chemistry in eucaryotic and procaryotic cell-systems are illustrated in this painting . Synthetic chemistry and pharmacology are linked in the development of the synthetic dyes . The chemical colours are often the same in dyes and drugs . The red colour in the pharmacology is associated with the antibiotic drugs as sulfonamides as well as with the staining of Gram-negative bacteria . The yellow colour is associated with the antibiotic drugs, quinolones and the Ziehl-Neelsen staining for tubercle bacillii . The blue colour is associated with the psychoactive drugs, phenotiazines, as well as with methylenblue staining and the staining of Gram-positive bacteria . These association and symbols have been used in this painting.

Commun Dis Public Health, 2002 Dec, 5(4), 289 - 98
Microbiological quality of take-away cooked rice and chicken sandwiches: effectiveness of food hygiene training of the management; Little CL et al.; During August 2001 a microbiological study of ready-to-eat cooked rice from take-aways and of chicken sandwiches made on the premises from sandwich bars was undertaken . The intention was to identify risk factors in the production, storage and handling of cooked rice and sandwiches, and to establish their effect on microbiological quality . Examination of cooked rice revealed that the majority of samples (87%; 442 of 508) were of satisfactory/acceptable microbiological quality; 50 (10%) were unsatisfactory, and 16 (3%) were of unacceptable quality due to Bacillus cereus and/or other Bacillus spp in excess of 10(5) cfu/g . The microbiological quality of cooked rice was associated with cuisine type (p < 0.00001), rice type (p < 0.01), cooking (p < 0.01), serving methods (p < 0.00001), and management food hygiene training (p < 0.01) . Examination of chicken sandwiches found that most (75%; 335 of 449) were of satisfactory/acceptable microbiological quality and 114 (25%) were unsatisfactory . Acceptable microbiological quality of sandwiches was associated with sandwich bars that had hazard analysis in place (p < 0.05) . Smaller businesses, as indicated by Local Authority Inspectors' Consumer at Risk scores, were more likely to have samples classified as unsatisfactory or unacceptable compared to larger businesses (p < 0.001) . The majority (90%) of premises had hand-washing facilities accessible and available for use, although only over half (55%) were correctly used as judged by the sampling officer . Where the manager of the premises had received some form of food hygiene training, food safety procedures such as the hazard analysis system were more likely to be in place (p < 0.0001).

Mem Inst Oswaldo Cruz, 2002 Dec, 97(8), 1157 - 63 Epub 2003 Jan 20.
Mtp-40 and alpha antigen gene fragment amplification for the detection of Mycobacterium tuberculosis in Colombian clinical specimens; Alfonso R et al.; In this study, the use of Mtp-40 and alpha antigen polymerase chain reaction (PCR) amplification fragments for the precise tuberculosis (TB) diagnosis was evaluated . One hundred and ninety two different samples were obtained from 113 patients with suspected TB . Mtp-40 and alpha antigen protein genes were amplified by the PCR technique and compared to both the "gold standard" (culture) test, as well as the clinical parameters (including a clinical record and X-ray film exam in 113 patients) . Thirty-eight of the 113 patients had a presumptive clinical diagnosis of TB; 74% being detected by PCR technique, 58% by culture and 44% by direct microscopic visualization . Weconclude that it is possible to use PCR as a suitable technique for the detection of any mycobacteria by means of the alpha antigen product, or the specific infection of Mycobacterium tuberculosis by means of the mtp-40 gene . This might be a good supporting tool in difficult clinical TB diagnosis and pauci-bacillary cases.

J Antimicrob Chemother, 2003 Feb, 51(2), 361 - 5
Enzymic degradation of a beta-lactam antibiotic, ampicillin, in the gut: a novel treatment modality; Harmoinen J et al.; Antibiotics can cause severe alterations in the gut microflora and promote diarrhoea and overgrowth of pathogenic bacteria . The present study investigated the potency of targeted recombinant beta-lactamase (TRBL) to degrade a beta-lactam antibiotic in the jejunum of fistula-operated beagles . We used different peroral doses of purified beta-lactamase (PenP) of Bacillus licheniformis in enteric-coated pellets together with intravenous ampicillin . Serum and jejunal samples were collected for ampicillin and beta-lactamase analysis . A dose-response effect of TRBL on ampicillin concentrations in the jejunal samples could be observed . The highest doses applied decreased the jejunal ampicillin concentrations to undetectable levels . In the serum samples, the ampicillin concentrations were not affected by the beta-lactamase dose used . Our results indicate that it may be possible to evolve a targeted treatment to degrade beta-lactam antibiotics intestinally and, thus, decrease antibiotic-induced adverse effects on the gut microflora.

Indian J Exp Biol, 2001 Jun, 39(6), 584 - 9
14C-{lignin}-lignocellulose biodegradation by bacteria isolated from polluted soil; Kumar L et al.; Four bacterial species {Branhamella catarrhalis (gram -ve), Brochothrix species (gram -ve), Micrococcus luteus (gram +ve) and Bacillus firmus (gram +ve)}, isolated from the soil polluted with cane sugar factory effluents, were found capable of growing on solid media supplemented with indulin AT (a polymeric industrial lignin) as sole C source . All the four species could metabolize cinnamic acid (a non-hydroxylated phenylpropanoid) as sole carbon source with significant suppression on addition of readily metabolizable carbon source (glucose) . However, Br . catarrhalis and Brochothrix sp . were capable of metabolizing ferulic acid, but could not do so on addition of glucose . Of the four species, Br . catarrhalis could evolve significant amount of 14CO2 from U-14C (lignin)-lignocellulose prepared from rice stalks (ca . 10% of the added radioactivity in 3 weeks), in addition to solubilization of another 11.7% radioactivity in culture filtrate . The other three species could not significantly evolve 14CO2, though a significant fraction of added 14C-lignin (6.1 to 11.2%) could be solubilized into culture filtrate, suggesting lack of ring-cleavage or other CO2 evolving mechanisms in these species.

Chemistry, 2002 Dec 2, 8(23), 5447 - 55
Chemoenzymatic syntheses of linear and branched hemithiomaltodextrins as potential inhibitors for starch-debranching enzymes; Greffe L et al.; Oligosaccharides embodying the S-maltosyl-6-thiomaltosyl structure have been readily synthesised by using convergent chemoenzymatic approaches . The key steps for the preparation of these molecules involved: 1) transglycosylation reactions of maltosyl fluorides onto suitable acceptors catalysed by the bacterial transglycosylase, cyclodextrin glycosyltransferase (CGTase), and 2) the SN2-type displacement of a 6-halide from acetylated acceptors by activated 1-thioglycoses . The target molecules, which were obtained in good overall yields, proved to be useful for investigating substrate binding in the active sites of several enzymes that act upon the alpha-1,6-linkage of pullulan and/or amylopectin . The compounds exhibit Ki values in the 2.5-1350 microM range with the different enzymes, and the highest affinity found by using these molecules was seen for the pullulanase from Bacillus acidopullulyticus . Both barley-malt limit dextrinase and pullulanase type II from Thermococcus hydrothermalis only recognised the longest linear thiooligosaccharide, while a branched heptasaccharide was the strongest inhibitor of pullulanase from Klebsiella planticola.

Sheng Wu Gong Cheng Xue Bao, 2002 Sep, 18(5), 578 - 82
{The analysis of Bacillus thuringiensis vegetative insecticical protein gene cloning and expression}; Cai QL et al.; Three kinds of Bacillus thuringiensis serotype-subsp . Leesis(H33) strain YBT-833, subsp . Aizawai(H7) strain YBT-1416 and subsp . Kurstaki(H3ab) strain YBT-1535, which were isolated by our lab, are chosen as original strain to clone vegetative insecticidal protein gene . Southern hybridization showed that vip genes are all localized at roughly 4-5 kb size-fractionated XbaI fragments of total DNA from YBT-833, YBT-1416 and YBT-1535 . Three subgenomic libraries containing the vip gene fragment, were constructed with pUC19 as vector . Then, three vegetative insecticidal protein gene vip83, vip14 and vip15 are obtained from the libraries through the methods of colony-blot-in-situ screening and enzyme-cut detection . Comparision of DNA sequence made out that only vip83 gene exist five different base pairs with known vip genes . Because the sequences of vip14 and vip15 are the same, two of the three genes, vip83 and vip14, were subcloned to shuttle vehicle pHT315 to get recombinant plasmids pBMB8901 and pBMB8902 in turn . The plasmids were separately transformed into vip Bt . receptors BMB171 and 4Q7 to obtain four engineered strains BMB8901-171, BMB8902-171, BMB8901-4Q7 and BMB8902-4Q7 . SDS-PAGE results indicated that all recombinant strains express 88 kD vegetative insecticidal protein . Bioassay also showed that the proteins of genes vip83 and vip14 both have certain toxicity to Lepidopteran insect larvae such as Heliochis armigera, Spodotera exigua and Plutella xylostella . While the toxicity of vip protein from four engineered strains to Plutella xylostellas are highest, whose LC50 value is 28.6, 31.6, 45.4 and 37.6 microL/mL respectively . This study will contributed to construct high efficacy and wide spectrum engineered strains on theory and reality.

J Mol Biol, 2003 Feb 14, 326(2), 353 - 69
Regulation of arginine biosynthesis in the psychropiezophilic bacterium Moritella profunda: in vivo repressibility and in vitro repressor-operator contact probing; Xu Y et al.; We report the cloning of the arginine repressor gene from the psychropiezophilic Gram-negative bacterium Moritella profunda, the purification of its product (ArgR(Mp)), the identification of the operator in the bipolar argECBFGH(A) operon, in vivo repressibility studies, and an in vitro analysis of the repressor-operator interaction, including binding to mutant and heterologous arginine operators . The ArgR(Mp) subunit shows about 70% amino acid sequence identity with Escherichia coli ArgR (ArgR(Ec)) . Binding of purified hexameric ArgR(Mp) to the control region of the divergent operon proved to be arginine-dependent, sequence-specific, and significantly more sensitive to heat than complex formation with ArgR(Ec) . ArgR(Mp) binds E.coli arginine operators very efficiently, but hardly recognizes the operator from Bacillus stearothermophilus or Thermotoga maritima . ArgR(Mp) binds to a single site overlapping the -35 element of argC(P), but not argE(P) . Therefore, the arrangement of promoter and operator sites in the bipolar argECBFGH(A) operon of M.profunda is very different from the organization of control elements in the bipolar argECBH operon of E.coli, where both promoters overlap the common operator and are equally repressible . We demonstrate that M.profunda argC(P) is about 44-fold repressible, whereas argE(P) is fully constitutive . A high-resolution contact map of the ArgR(Mp)-operator interaction was established by enzymatic and chemical footprinting, missing contact and base-specific premodification binding interference studies . The results indicate that the argC operator consists of two ARG box-like sequences (18bp imperfect palindromes) separated by 3bp . ArgR(Mp) binds to one face of the DNA helix and establishes contacts with two major groove segments and the intervening minor groove of each ARG box, whereas the minor groove segment facing the repressor at the center of the operator remains largely uncontacted . This pattern is reminiscent of complex formation with the repressors of E.coli and B.stearothermophilus, and suggests that each ARG box is contacted by two ArgR subunits belonging to opposite trimers . Moreover, the premodification interference patterns and mutant studies clearly indicate that the inner, center proximal halves of each ARG box in the M.profunda argC operator are more important for complex formation and repression than the outermost halves . A close inspection of sequence conservation and of single base-pair O(c)-type mutations indicate that the same conclusion can be generalized to E.coli operators.

Carbohydr Res, 2003 Feb 14, 338(5), 415 - 21
Characterizing the pH-dependent stability and catalytic mechanism of the family 11 xylanase from the alkalophilic Bacillus agaradhaerens; Poon DK et al.; The xylanase, BadX, from the alkalophilic Bacillus agaradhaerens was cloned, expressed and studied in comparison to a related family 11 xylanase, BcX, from B . circulans . Despite the alkaline versus neutral conditions under which these bacteria grow, BadX and BcX both exhibit optimal activity near pH 5.6 using the substrate o-nitrophenyl beta-xylobioside . Analysis of the bell-shaped activity profile of BadX yielded apparent pK(a) values of 4.2 and 7.1, assignable to its nucleophile Glu94 and general acid Glu184, respectively . In addition to having an approximately 10-fold higher k(cat)/K(m) value with this substrate at pH 6 and 40 degrees C, BadX has significantly higher thermal stability than BcX under neutral and alkaline conditions . This enhanced stability, rather than a shift in its pH-optimum, may allow BadX to hydrolyze xylan under conditions of elevated temperature and pH.

Med Parazitol (Mosk), 2002 Oct-Dec, (4), 23 - 7
{The duration of action of Bacillus thuringiensis spp . israelensis and Bacillus sphaericus after encapsulation by infusoria Tetrahymena pyriformis}; Ganyshkina LA et al.; The study of the duration of the insecticidal activity of a mixture of infusoria with each of these bacteria revealed differences in the pattern of their relations during their long contact . T . pyriformis may be used for the prolongation of its insecticidal action if Bti is applied . An increase in the duration of this action depends on the viability of tetrahymena in the experimental flasks and on the presence of free spores and crystals of Bti . The prolongation of the action may be associated with the multiplication of this type of bacteria in the digestive vacuoles of infusoria . The use of Bsph in the mixture with T . pyriformis caused no increase in the insecticidal activity of bacteria . Tetrahymena increased the duration of Bsph and even reduced the period of its larvicidal activity . Tetrahymena used a sporocrystalline complex of Bsph as food and reduced its water concentration.

Wei Sheng Wu Xue Bao, 2002 Apr, 42(2), 175 - 80
{Analysis of function of the 452 bp sequence proceeding the pgiB gene in Bacillus stearothermophilus}; Chen L et al.; By random mutagenesis with chemical mutagen N-methyl-N'-nitro-N-nitrosoguanidine, a shuttle promoter probe vetor mutant pPGV5 (tr65) was obtained . A single site mutation from G to T was discovered at nucleotide + 238 of kan gene encoding kanamycin nucleotidyltransferae by sequence analysis . Using the gene pheB that encodes thermostable catechol 2'3-dioxygenase from Bacillus stearothermophilus FDTP-3 as the reporter gene, a transcriptional fusion plasmid pPGVPB452 was constructed and electroportated into B . stearothermophilus . By analyzing the reporter protein, it proved that the 452 bp sequence proceeding the coding region of phosphoglucoisomerase pgiB gene from B . stearothermophilus T521, which contained a putative promoter by sequence analysis, but has no transcriptional activity in B . stearothermophilus.

Wei Sheng Wu Xue Bao, 2002 Apr, 42(2), 169 - 74
{Characterization of cry gene and broad spectrum against lepidopteran of Bacillus thuringiensis subsp . colmeri 15A3}; Chen Y et al.; Bacillus thuringiensis wild type strain 15A3 belongs to subspecies colmeri serotype H-21 . RFLP and PCR analysis show that it contains six types of ICP genes: cry1Aa, cry1Ac, cry1Ca, cry1D, cry1I and cry . The sequence of the 1.45 kb N-terminal fragment of cry1Aa differed from that of published . SDS-PAGE showed that the crystal consists of proteins with molecular weight about 130, 79, 70, 65, 51 and 45 kD . Strain 15A3 didn't sysnthesize heat-stable beta-exotoxins according to test of house fly aberration . The 1.2 tons fermentative production exhibited high toxicity against three lepidopteran pests: H . armigera, S . exigua and H . cunea . It was proved that wild type strain can produce a broad specturm of ICP.

Wei Sheng Wu Xue Bao, 2002 Jun, 42(3), 335 - 40
{Study on amount and stability of cry1D and cry1Ab mRNA in Bacillus thuringiensis subps aizawai}; Chang L et al.; cry1Ab, cry1C and cry1D are three typical cry1 class genes encoding protoxins in Bacillus thuringiensis subsp . aizawai strain HD-133 . The expression of cry1D is obviously different from that of cry1Ab in this strain . Amount and stability of cry1D and cry1Ab mRNA in HD-133 were investigated . Northern blotting analysis showed that cry1D mRNA was more stable than cry1Ab mRNA, however, cry1D mRNA formed later 3 h and less 3.7 times than cry1Ab mRNA during the mid-phase of sporulation . Halve lives of cry1D mRNA was 18 min, but halve lives of cry1Ab mNRA was 14 min . Moreover, cry1Ab mRNA could keep stable and large amount during the post-phase of sporulation . This suggested that difference of transcriptional efficiency and initiate time might be main reason for the difference of expression of cry1Ab and cry1D.

Wei Sheng Wu Xue Bao, 2002 Feb, 42(1), 76 - 80
{Studies on enzymatic synthesis of cephalexin by immobilized penicillin acylase by polyacrylonitrile fibres}; Chen H et al.; The extracellular penicillin acylase from Bacillus megaterium was immobilized by coupling to derivatives of polyacrylonitrile fibres . The apparent activity of the immobilized enzyme was about 153 U/g (wet weight) . The optimal pH and temperature were 6.5 and 40 degrees C for synthesis of cephalexin by penicillin acylase, respectively . Whe the concentration of 7-ADCA was 4% and the ratio of PGME and 7-ADCA and 1:2, the average velocity of synthesis reaction was highest . The optimal supplied amount of immobilized penicillin acylase was 1.125 g/g 7-ADCA. . The apparent Michaelis constant for 7-ADCA was 0.162 mol/L and for PGME was 0.364 mol/L, Vmax was 0.0462 mol.L-1.min-1 at 30 degrees C and pH6.5 . The remained activity was about 83.9% after operating 50 times.

Wei Sheng Wu Xue Bao, 2002 Feb, 42(1), 45 - 9
{The cloning of plasmid replicon ori165 from Bacillus thuringiensis subsp . thenebrionis}; Wei F et al.; An EcoRI DNA fragment containing the plasmid replicon, named ori165, was cloned from the plasmids of B . thuringiensis subsp . tenebrionis strains YBT-1765 . The replicon ori165 was the first replicon cloned from this subspecie . Its size is approximately 11 kb . The replicon was minimized to an 8 kb fragment by deletion analysis . Southern hybridization exhibited that ori165 was coming from the plasmid detected with the highest molecular weight . A shuttle vector pBMB6071 based on ori165 was constructed . The stabilities of pBMB6071 in different B . thuringiensis host had great discrepancy . In strain 4Q7, a crystal-negative derivative of B . thuringiensis subsp . isrealensis, the stability was 100% . ori165 was compatible with ori1030 and ori2062 in strain BMB171, a crystal-negative derivative of B . thuringiensis subsp . kurstaki.

Wei Sheng Wu Xue Bao, 1999 Aug, 39(4), 381 - 4
{Detecting microorganisms in soil by PCR and molecular hybridization}; Ding Z et al.; PCR and molecular hybridization provide new methods to detect microorganisms in environment . A simple, sensitive protocol was developed to detect Bacillus thuringiensis HD-1 in soil by PCR and Southern Blotting in this study . Methods of extracting DNA from soil and purification were compared . By a modified total DNA extraction from soil and purification, followed by PCR and Southern Blotting, strain HD-1 released into soil can be detected with a sensitivity of 2.2 x 10(2) cells/g soil . By using other specific primers, this detection protocol can be widely used in the research of ecology and risk assessment of genetically engineered microorganisms released into environment.

Wei Sheng Wu Xue Bao, 1999 Aug, 39(4), 344 - 9
{Purification and characterization of a novel antifungal peptide APS-1 produced by Bacillus cereus}; Pei Y et al.; In previous study, we isolated an antagonist Bacillus cereus strain: S-1 from cotton plant . In field experiments, this bacterium was shown strong inhibition to several plant diseases . In this paper, we reported the purification of the antifungal substance produced by the bacterium and its properties . After the steps of acid precipitation, methanol and ethy lether extraction, Sephadex G100 and DEAE52 column chromatography, the antifungal material was purified . The purified material had absorption peak at 275 nm, and was exhibited negative in biuret color reaction . However after hydrolyzed with HCl, this substance shown positive in the same reaction . Amino acid analysis to the hydrolysate of APS-1 showed that APS-1 was composed of Glu, Asp, Tyr, Ser, Thr, Pro, Leu, Ile, Val and an unknown amino acid . Combining with its pratial resistance to proteinases, it was suggested that this antifungal material was a cyclic peptide . This peptide, named APS-1, with strong inhibition on the germination of spores of the phytopathogens tested, was shown high stability against ultraviolet radiation and heat . APS-1 may have potential role in plant diseases biological control.

Wei Sheng Wu Xue Bao, 1999 Dec, 39(6), 515 - 20
{New coleopterancidai strains of Bacillus thuringiensis and production of coleopterancide}; Gao M et al.; Two coleopterancidal strains of Bacillus thuringiensis, YM-03 and SHQ11-10, were isolated from soil in China . H-Serotype of YM-03 was H8a8b, and H-Serotype of SHQ11-10 has not been identified . Both of them produced flat bipyramid-like crystals which consisted of 68-70 kD and 65 kD protein respectively . Bioassay indicated that these two Bt strains were very active on the larva of Leptinotarsa decemlineata and Plagioder aversicolora . They grew prosperously in fermentation . Powder of YM-03 product was high effect against colorado potato beetle in the field experiment.

Wei Sheng Wu Xue Bao, 1999 Jun, 39(3), 260 - 3
{Studies on reduction of Au3+ by bacteria for preparing gold catalyst}; Liu Y et al.; The strain D01, screened from different source bacterial strains, has relatively strong ability of adsorbing Au3+ . The strain D01 was identified as Bacillus megatherium D01 and still grew well in the medium containing 600 mg/L Au3+ . The results of electrochemical reaction indicated that the strain D01 had a strong ability of reduction . It could reduce the Au3+/alpha-Fe2O3(precursor of the gold catalyst) to highly dispersive Au0/alpha-Fe2O3 catalyst which could catalyze the reaction of CO + O2-->CO2.

Wei Sheng Wu Xue Bao, 1999 Oct, 39(5), 469 - 74
{The effects of reactive oxidants on Bacillus thuringiensis parasporal crystals}; Wang W et al.; The effects of hydrogen peroxides(H2O2) and hydroxyl radicals(.OH) on the purified parasporal crystals from Bacillus thuringiensis var . Kurstaki HD-1 were studied by SDS-PAGE and bioassay . The results showed that to some extent H2O2 and .OH produced a direct and deleterious effect . Hydroxyl radicals from the system Fe(II)-EDTA-H2O2 indicated more serious damage than hydrogen peroxides . There was a 44.5% loss in activity against 12-h-old larvae of Heliothis armigera when parasporal crystals were treated with 0.3% H2O2 only . However, 85.2% activity lost when parasporal crystals were treated with 0.3% H2O2 in the presence of iron(II) and EDTA.

Wei Sheng Wu Xue Bao, 1999 Oct, 39(5), 426 - 9
{Location of the binary toxin gene of Bacillus sphaeriucs and some biological characteristics of its asprogenous mutants}; Liu Y et al.; Location of the binary toxin gene of Bs C3-41 and some biological characteristics of its asporogenous mutants which obtained by DES treatment were studied in this paper . Mutant C4, L5 with blocked at the stage II of sporulation course were not possessed of any crystalline inclusion, but the binary toxin proteins were detected in their cells by SDS-PAGE analysis . Another mutant G5 which blocked at the stage III of sporulation course formed parasporal crystal in the cells, and its toxicity to larvae of Culex quinquefasciatus was about 50 times higher than those of mutant C4 and L5 . The result of curing plasmid and Southern blot analysis confirm the binary toxin gene of Bs C3-41, Bs 1691 and Bs-10 locate on the chromosome.

Wei Sheng Wu Xue Bao, 1999 Apr, 39(2), 154 - 9
{The identification of 5 new serotypes of Bacillus thuringiensis from soil in China}; Li R et al.; Five new strains of Bacillus thuringiensis(Bt) from soil in China were classified . 5 new H-serotypes: H42, H43, H56, H60 and H62 were identified . There were some distinction in the morphology, biochemical reaction and toxicity against species of insect . Five new Bt subspecies were identified as follows: Strain YGd22-03 Bacillus thuringiensis subsp . jinghongiensis, H42; Strain KK31-01 Bacillus thuringiensis subsp . guiyangiensis, H43; Strain SCg04-02 B . thuringiensis subsp . rongseni, H56; Strain NXP15-04 B . thuringiensis subsp . pingluonsis, H60; Strain HZ39-04 B . thuringiensis subsp . zhaodongensis, H62.

Wei Sheng Wu Xue Bao, 1999 Apr, 39(2), 114 - 9
{Preliminary study of P19 gene from Bacillus thuringiensis subsp israelensis}; Liu Z et al.; The P19 gene and cyt1A gene were obtained by PCR with 9.7 kb HindIII fragment containing P19 gene and cyt1A gene from 72 MD plasmid of Bt as a template . Digested PCR products were ligated to expression vector pUHE24 and transformed into E . coli XL-1 . Three clones, LZ19 harboring P19 gene, LZcyt1A harboring cyt1A gene, LZ19A harboring P19 gene and cyt1A gene, were screened . The growth cure of cloned strains were determined under IPTG induction . The result showed that pLZ19 did not affect the growth of E . coli, pLZcyt1A is typically Lethal for E . coli, Lethal initial efficiency of pLZ19A is much higher than that of pLZcyt1A . Probably, this was a result that P19 gene enhanced initial expression of cyt1A gene in E . coli.

Wei Sheng Wu Xue Bao, 1999 Feb, 39(1), 29 - 35
{Cloning and expression of the binary toxin genes of Bacillus sphaericus C3-41 in a crystal minus B . thuringiensis subsp . israelensis}; Yuan Z et al.; Bacillus sphearicus C3-41, belonging to serotype H5a5b, is ahighly toxic strain isolated from a mosquito-breeding site in China . It has been shown that it had a higher toxicity against Culex spp . than the reference strain 2362 at the laboratory and field conditions . Using synthetic oligonucleotides designed on the basis of the binary toxin gene sequence of 2362, a 1.1 kb DNA fragment was produced and the genetic library was prepared from a HindIII digest of total DNA from C3-41 . One colony containing the 3.5 kb HindIII fragment was selected for further studies . Sequence analysis revealed that this 3.5 kb Hind DNA fragment was composed of 3479 nucleic acids and the sequence of the binary toxin gene of C3-41 is completely identical to that of strain 2362 . The toxin genes have been transferred into a nontoxic crystal-minus strain of Bacillus thuringiensis subsp . israelensis . The recombinant strains could express the binary toxin of B . sphaericus as crystaline structures during their sporulation . The expression products of the recombinants have a highly toxicity to susceptible Culex pipiens subsp . quinquefasciatus and no toxicity to resistant larvae(Culex pipiens subsp . pipiens) from France.

Acta Crystallogr D Biol Crystallogr, 2003 Feb, 59(Pt 2), 348 - 9 Epub 2003 Jan 23.
Crystallization and preliminary X-ray diffraction studies of alpha-cyclodextrin glucanotransferase isolated from Bacillus macerans; Choe HW et al.; Single crystals of alpha-cyclodextrin glucanotransferase isolated from Bacillus macerans have been grown with polyethylene glycol 6000 as a precipitating agent by sitting-drop vapour diffusion at room temperature . The crystals were suitable for X-ray analysis and diffracted to at least 2.0 A (space group P2(1)2(1)2(1)), with unit-cell parameters a = 66.79 (2), b = 79.66 (1), c = 141.16 (1) A . Assuming the asymmetric cell to be occupied by a monomer of 74 kDa, the unit cell contains 42.6% solvent with a crystal Volume per protein mass, V(M), of 2.53 A(3) Da(-1).

Acta Crystallogr D Biol Crystallogr, 2003 Feb, 59(Pt 2), 341 - 2 Epub 2003 Jan 23.
Crystallization and preliminary X-ray analysis of a thermostable pectate lyase PL 47 from Bacillus sp . TS 47; Nakaniwa T et al.; The thermostable pectate lyase PL 47 from Bacillus sp . TS 47, with a molecular weight of 50 kDa, was crystallized by the hanging-drop vapour-diffusion method using 2-propanol and polyethylene glycol 4000 as precipitants . The crystals belong to the trigonal space group P3(1)21, with unit-cell parameters a = b = 58.8, c = 229.7 A, gamma = 120 degrees . The calculated V(M) based on one molecule per asymmetric unit is 2.30 A(3) Da(-1) . A native data set from a frozen crystal was collected to 1.8 A resolution using synchrotron radiation at SPring-8 . A molecular-replacement solution was obtained using the structure of pectate lyase from B . subtilis as a model.

J Exp Bot, 2003 Feb, 54(383), 835 - 44
Transgenic loblolly pine (Pinus taeda L.) plants expressing a modified delta-endotoxin gene of Bacillus thuringiensis with enhanced resistance to Dendrolimus punctatus Walker and Crypyothelea formosicola Staud; Tang W et al.; A synthetic version of the CRY1Ac gene of Bacillus thuringiensis has been used for the transformation of loblolly pine (Pinus taeda L.) using particle bombardment . Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants . Expression vector pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) CRY1Ac coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator sequences, and the neomycin phosphotransferase II (NPTII) gene controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment . The transformed tissues were proliferated and selected on media with kanamycin . Shoot regeneration was induced from the kanamycin-resistant calli, and transgenic plantlets were then produced . More than 60 transformed plants from independent transformation events were obtained for each loblolly pine genotype tested . The integration and expression of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern hybridization, by Northern blot analysis, and by Western blot analysis . Effective resistance of transgenic plants against Dendrolimus punctatus Walker and Crypyothelea formosicola Staud was verified in feeding bioassays with the insects . The transgenic plants recovered could represent a good opportunity to analyse the impact of genetic engineering of pine for sustainable resistance to pests using a B . thuringiensis insecticidal protein . This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform.

J Pharm Pharm Sci, 2002 Sep-Dec, 5(3), 272 - 8
Response surface optimization of the critical medium components for the production of alkaline protease by a newly isolated Bacillus sp; Adinarayana K et al.; PURPOSE: Optimization of the fermentation medium for maximum alkaline protease production was carried out using a new strain, Bacillus Sp . PE-11 . METHODS: The carbon source (glucose), the nitrogen source (peptone) and salt solution were selected to optimize . A 2(3 )full factorial composite experimental design and response surface methodology were used in the design of experiments and in the analysis of results . This procedure limited the number of actual experiments performed while allowing for possible interactions between the three components . RESULTS AND DISCUSSION: The optimum values for the tested variables for the maximum alkaline protease production were; glucose 7.798 (g/L), peptone 9.548 (g/L) and salt solution 8.757% . The maximum alkaline protease production was 4,98,123 PU/L . This method was efficient; only 20 experiments were necessary to assess these conditions, and model adequacy was very satisfactory, as the coefficient of determination was 0.941 . CONCLUSIONS: In the work, we have demonstrated the use of a central composite factorial design by determining the conditions leading to the high yield of enzyme production . Thus, smaller and less time consuming experimental designs could generally suffice for the optimization of many fermentation processes.

Wei Sheng Wu Xue Bao, 2001 Aug, 41(4), 475 - 80
{Immobilization of penicilin G acylase on polyacrylonitrile fiber}; Xian H et al.; The immobilization of Penicillin G Acylase from Bacillus megaterium by glutaraldehyde crosslinking on the partially acid-hydrolyed polyacrylonitrile fiber was studied . When the amount of--NH2 on fiber were 690 mumol/g and the moisture in the fiber was 64%, and the content of enzyme protein immobilized on fiber was more than 100 mg/g . The activity of 2300 IU/g was obtained with 30% of overall yield and 56% of binding efficiency . The immobilization yield was markedly influenced by ratio of the amount of free enzyme used to the weight of the fiber . The half-life of storage stability of immobilized PGA at room temperature was 130 days . The immobilized PGA kept 80% of the initial activity after 20 cycles of operation in 10% of PGK(W/V) in 0.05 mol/L phosphate buffer, pH 8.0, at 37 degrees C and an enzyme load of 150 IU/g(PGK) and 10 g(PGK) for per cycle of operation . The hydrolysis conversion of PGK in the range of 2.5%-12.5% (W/V) were over 98% for the immobilized PGA . The operation stability of immobilized PGA treated with DTT was better than that of immobilized PGA untreated.

Wei Sheng Wu Xue Bao, 2001 Aug, 41(4), 463 - 8
{Purification and characterization of 2-carbonyl reductase from marine bacteria Bacillus sp.}; Li X et al.; ANADPH-dependent 2-Oxoaldehyde reductase was isolated and purified from a marine bacteria Bacillus sp . The purification procedure involved ammonium sulfate fractionation and Q Sepharose FF, Hydroxyapatite, Sephadex G-100 column chromatographies . The specific activity of the purified enzyme was increased by 141.1 folds over crude extract and the recovery yield was 11.4% . 2-Oxoaldehyde compounds were found to be speciall good substrates . The optimum pH of the enzyme activity was 6.2-6.6 . The Km coefficient for 3-deoxyglucosone was 2.5 mmol/L . The molecular weight of the enzyme was estimated to be 33 kD The enzyme activity is stable below 30 degrees C and pH 5.0-8.0 . EDTA, beta-mercaptoethanol and dithiothreitol enhanced the enzyme activity . On the other hand, the enzyme activity was partially lost by idoacetic acid or N-ethylmaleimide.

Wei Sheng Wu Xue Bao, 2001 Oct, 41(5), 611 - 6
{Fermentation conditions for production of alkaline elastase by alkaliphilic Bacillus XE22-4-1}; Xiao C et al.; A extracellular alkaline elastase producing bacterial strain Bacillus XE22-4-1, with the optimum pH of 10.0 for its growth, was isolated and screened from alkali lake of Tibet . 2% glucose and 0.25% yeast extract are respectively the appropriate carbon and nitrogen source on elastase production . Soymeal can promote elastase production . The fermentation conditions in a 2 L stirred tank were investigated . The results have revealed that the dissolved oxygen is the most effective factor on elastase production . The maximum elastolytic activity reached 266 u/mL among 48 hours of fermentation by combination of enhancing aeration and changing the mixing speed.

J Infect Dis, 2003 Jan 15, 187(2), 270 - 8 Epub 2003 Jan 06.
Whole blood bactericidal activity during treatment of pulmonary tuberculosis; Wallis RS et al.; The timely evaluation of new drugs that can be used to shorten tuberculosis (TB) treatment will require surrogate markers for relapse . This study examined bactericidal activity against intracellular Mycobacterium tuberculosis in whole blood culture (whole blood bactericidal activity; WBA) during TB treatment . In the absence of chemotherapy, immune mechanisms in patient blood resulted in bacteriostasis, whereas administration of oral chemotherapy resulted in bacillary killing . Total WBA per dose was greater during the intensive phase of treatment than during the continuation phase (mean, -2.32 vs . -1.67 log(10) cfu-days, respectively; P<.001) . Cumulative WBA throughout treatment was greater in subjects whose sputum cultures converted to negative by the eighth week of treatment than in those for whom conversion was delayed (mean, -365 vs . -250 log(10) cfu-days; P=.04) and correlated with the rate of decrease of sputum colony-forming unit counts during the first 4 weeks of treatment (P=.018), both of which are indicative of prognosis . These findings indicate that measurement of WBA may have a role in assessing the sterilizing activity of new anti-TB drugs.

J Infect Dis, 2003 Feb 1, 187(3), 513 - 7 Epub 2003 Jan 24.
Lipoarabinomannan-reactive human secretory immunoglobulin A responses induced by mucosal bacille Calmette-Guérin vaccination; Brown RM et al.; The ability of 17 recombinant mycobacterial proteins, native antigen 85 complex, lipoarabinomannan (LAM), and Mycobacterium tuberculosis lysate to detect antibody responses induced by bacille Calmette-Guerin (BCG) vaccination and active tuberculosis infection were studied in enzyme-linked immunosorbent assays . Only LAM-reactive serum immunoglobulin G responses were significantly increased in both BCG-vaccinated patients and patients with active tuberculosis (P<.05), and oral BCG vaccination also induced significant increases in LAM-reactive secretory immunoglobulin A (P<.05) . LAM-reactive antibody assays can serve as markers of humoral and mucosal immunity in future trials of BCG and newer attenuated mycobacterial vaccines.

Lepr Rev, 2002 Dec, 73(4), 386 - 8
Relapse of lepromatous leprosy after WHO/MDT with rapid bacterial growth; Oliveira ML et al.; The authors report a case of relapse in a lepromatous patient 6 years after he had been cured by MDT/WHO/24 doses . The atypical aspect emphasized in this case is the bacterial load increase in a short period of time of 1 year after the smear count was negative, and the case reinforces the importance of patient education on release . No leprosy cases were identified in the patient's close contacts . It seems that relapse was a result of bacillary persistence, since a significant improvement was noted in relapsed lesions after two doses of MDT/WHO.

Wei Sheng Wu Xue Bao, 1998 Aug, 38(4), 310 - 2
{Purification and properties of alkaline cellulase from alkalophilic Bacillus N6-27}; Tian X et al.; The alkaline cellulase produced by alkalophilic Bacillus sp . N6-27 was purified to electrophoresis homogeneity by (NH4)2SO4 fractionation, Sepharose CL-4B hydrophobic interaction chromatography, Bio-gel P-150 chromatography . The molecular weight and pI determined by SDS-PAGE and by PAGE-IEF were 94,000 and 4.2, respectively . The optimum temperature and pH for the enzymatic catalysis were 55 degrees C and 8.5, respectively . The enzyme activity was stable under 50 degrees C and in the pH range of 6-11 . The substrate was carboxymethylcellulose (CMC) . The enzyme activity was strongly inhibited by Fe2+, Cu2+ and Hg2+.

Wei Sheng Wu Xue Bao, 1998 Aug, 38(4), 300 - 3
{Enzymatic synthesis of cephradine}; Cui F et al.; Cephradine was synthesized by gamma-alumina-immobilized form of the penicillin G acylase of Bacillus megaterium with D-phenyglycine methylester hydrochloride (CH DGME.HCl) as acyl donor and 7-aminodeacetoxycephalosporanic acid (7-ADCA) as acyl acceptor . 0.1 g of 7-ADCA was dissolved by adding 2.5 ml of distilled water and about 0.25 ml of 2 mol/L NaOH in a 25 ml flask . To the solution, after 0.25 g of CHDGME.HCl was added, 0.1 mol/L phosphate-0.05 mol/L citric acid buffer, pH 7.5 was added to result in a volume of 5 ml with pH 7.5 Then 1 g(220 IU) of immobilized enzyme was added . The flask was shaken on a rotary shaker at 110 r/min and 25 degrees C for 5 h . The conversion rate of 7-ADCA was 81% . In an expanded experiment in 500 ml of reactive volume, 11.8 g of cephradine was obtained from 10 g of 7-ADCA . The conversion rate of 7-ADCA was 80% with about 87% yield of cephradine . Enzymatic synthesis was inhibited in varying degrees by phenylacetic acid, phenoxyacetic acid and cephalosporin G.

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi, 1998 Mar, 15(1), 38 - 40
{Study on immobilized cells for producing alpha-amylase by using polyving alcohol as the carrier(II): The effect of fermentating conditions on the ability producing alpha-amylase of the cells immobilized with polyving alcohol as the corrier and continuous fermentation of the immobilized cells in CSTR}; Liu Z et al.; The effects of fermentating conditions on the ability of immobilized cells with PVA as carrier for producing alpha-amylase were studied . The continuous fermentation with the immobilized cells were tested in continuous flow stirred tank reactor (CSTR) . The results showed that the adaptability of the immobilized Bacillus substilis to pH increased after immobilization . In CSTR, the immobilized cells can be fermentated continuously for 360 hrs and the activity of alpha-amylase can be kept on the level of about 170 u/ml.

Wei Sheng Wu Xue Bao, 1998 Jun, 38(3), 204 - 7
{Studies on some properties of the immobilized penicillin acylase by polyacrylonitrile fibres}; Han H et al.; The extracellular penicillin acylase from Bacillus megaterium was immobilized by coupling to derivatives of polyacrylonitrile fibres . The apparent activity of the immobilized penicillin acylase was about 2,000 u/g (dry weight) . The optimal pH and temperature were 9.0 and 50 degrees C for hydrolysis of penicillin G, respectively . The immobilized enzyme was stable in the pH range of 5.5-10.3, and at the temperature below 50 degrees C . The apparent Michaelis constant Ka and Vm of the immobilization enzyme were 1.33 x 10(-2) mol/L and 2.564 mmol.min-1, respectively . The inhibition constant of phenylacetic acid acted as a competitive inhibitor for The immobilization enzyme was 0.16 mol/L . The remained activity was about 80% after operating 20 times.

Wei Sheng Wu Xue Bao, 1998 Jun, 38(3), 181 - 5
{Selection of drug-labelled Bacillus sphaericus and studies on transformation and expression of foreign DNA}; Zhang P et al.; NTG was used to make chemical mutation for Bacillus sphaericus, RifR and SmR labelled strains were selected, which could resist drug as much as 100 u/ml . The resistance to drug was stably inherited . The RifR strain was used as recipient and the plasmid containing the lysostaphin gene was transfered into it by protoplasts . Results showed that the lysostaphin gene could be expressed stably at high level in Bacillus sphaericus and the lysostaphin activity was about 122 u/ml medium after shaking culture.

Wei Sheng Wu Xue Bao, 2001 Feb, 41(1), 76 - 81
{Cloning and DNA sequencing of Bacillus pumilus endo-1,4-beta-glucanase gene}; Yang Z et al.; Bacillus pumilus S-27 screened from marine environment produces an extracellular endo-1,4-beta-glucanase . The enzyme has its optimal activity at pH 5.0-7.0 and 55 degrees C and still retained 79.0% of it at pH 9.0 . The gene encoded this endo-1,4-beta-glucanase was cloned and sequenced . The structural gene contained an open reading frame of 1980 bp, corresponding to 660 amino acids, the amino acid sequence of this enzyme is very close to that of an EG of C . cellulovorans and an EG of Bacillus sp . KSM-522, all belong to the cellulase family E2.

Wei Sheng Wu Xue Bao, 2001 Jun, 41(3), 357 - 62
{Synthesis of galacto-oligosaccharides by immobilized Bacillus stearothermophilus}; Chen S et al.; Bacillus stearothermophilus was embedded in sodium alginate, chitosan, gelatin respectively and utilized for galacto-oligosaccharides (GOS) production . By comparing with each other in the enzyme recovery, optimal reaction conditions, yields of GOS and mechanical strength of the carriers, the gelatin was selected as better carrier for immobilization . pH, temperature, lactose concentration, lactose conversion and mass transfer resistance of carrier significantly influenced the Yield of GOS . The maximum yield of GOS was 31.2% in a stirred reactor with lactose concentration at 60% . It remained about 88% after 96 h (8 batches) reaction . A packed bed was employed for a continuous reaction at space velocity 0.09 h-1, where GOS yield and reactor productivity was 31.5% and 17.4% respectively . It lost about 20% of its original yield of GOS after 140 h reaction . Products were separated by active charcoal columm . The chemical structure of tetrasaccharide identified by 13C-NMR was beta-D-Gal-(1-->3)-D-Gal-(1-->6)-Dal-(1-->4)-D-Glu.

Wei Sheng Wu Xue Bao, 2001 Jun, 41(3), 304 - 9
{Transcriptional activator PlcR regulate the expression of multiple genes in Bacillus cereus}; Wang L et al.; Food-poisoning and some diseases of human can be caused by potential pathogenic factors including phospholipases of c-type, hemolysins, enterotoxins, emetic toxin and others from Bacillus cereus which is an opportunistic pathegon . Generally, the expression of pathogenic genes is completed by a co-regulation in many bacteria . The isolation of a mutant, displaying significant reducing expression of lecithinase and general proteolytic activities, from a transposon-induced bank of B . cereus type strain ATCC14579T were described . Sequencing data revealed that a gene with high homology to transcriptional activator (plcR) of B . thuringiensis was inactivated by insertion in this mutant . The results suggest that plcR regulates the expression of multiple genes in B . cereus.

Wei Sheng Wu Xue Bao, 2001 Jun, 41(3), 293 - 7
{Identification and location of the toxin protein genes in 56 Bacillus thuringiensis isolates}; Zhong X et al.; Polymerase chain reaction was performed to screen Bacillus thuringiensis isolates for cryI, cryIII, and cryV genes with cryI-, cryIII-, and cryV-specific primers respectively . The results indicated that, in the 56 Bt isolates examined, 7 isolates contain cryI gene, 2 isolates contain cryIII gene, and 21 isolates contain both cryI and cryV genes . The cryV gene was located by Southern hybridization in the large plasmid of about 150 MD molecular mass.

Shi Yan Sheng Wu Xue Bao, 2000 Dec, 33(4), 357 - 65
{The somatic embryogenesis and establishment of transformation experiment system in Larix principis-Rupprechtii}; Qi LW et al.; Larix principis-Rupprechtii is one of the superior afforestation forest trees growing in north China . Embryogenic cultures were initiated from immature zygotic embryos of Larix principis-Rupprechtii on S culture medium containing 2, 4-D 0-2.2 mg/L, KT and BA each at 0-0 . 8 mg/L . Embryogenic calli were subcultured and multiplicated on S + B culture medium containing dropping off each hormone concentration . We set up 33 steady-going embryogenic cell lines; We studied on the growth stage and genotype differences of every embryogenic cell lines; and Finded more than 10 high-frequency somatic embryogenesis cell lines such as 2K, 2T, 2I, 2J, 3C etc. . The number of 2T somatic embryos reaches 314/per gram of embryogenic tissue and the number of 3C somatic embryos is 185/per gram of embryogenic tissue . The re-induction method of Larix principis-Rupprechtii from somatic embryos was used to produce renewable embryogenic cultures and steady-going embryogenic cell lines effectively . Mature somatic embryos can germinate and develop further into plantlets when they are isolated and cultured on a hormone-free WPM culture medium . The regeneration plantlets were obtained . Furthermore, the transformation with a truncated gene of Bacillus thuringensis (B . t) were carried out, the PCR showed positive results, because of this, embryogenic cell line of Larix principis-Rupprechtii can be used for transformation experiments to support further breeding in forestry.

Wei Sheng Wu Xue Bao, 2001 Apr, 41(2), 204 - 8
{Studies on the immobilized penicillin acylase on polymer beads}; Han H et al.; The extracellular penicillin acylase from Bacillus megaterium was immobilized on oxirane group of Eupergit C beads . The apparent activity of the immobilized enzyme was about 1400 u.g-1 (dry weight) . The optimal pH and temperature were 8.0 and 55 degrees C for hydrolytic reaction of penicillin G, respectively . The immobilized enzyme was stable in the pH range of 6.0-8.5 and at temperature below 45 degrees C . The apparent Michaelis constant for penicillin G was inhibition constant of phenylacetic acid as competitive 2 x 10(-2) mol.L-1 and Vm was 1.33 mmol.g-1 min-1 (dry weight) at 37 degrees C and PH8.0 . The inhibitor and 6-APA as non-competitive inhibitor were 2.8 x 10(-2) mol.L-1 and 0.125 mol.L-1 for the immobilized enzyme at pH 8.0 and 37 degrees C, respectively . The remained activity of the immobilized enzyme was about 80% after operating 200 times for hydrolysis of penicillin G to 6-APA, and the average yield of 6-APA was 89.48%.

Wei Sheng Wu Xue Bao, 2001 Apr, 41(2), 148 - 54
{Detection of enterotoxin genes in Bacillus thuringiensis by PCR}; Yuan Z et al.; The presence of hemolysin HblA (hblA), enterotoxin BceT (bceT) and enterotoxin S (entS) genes from 45 strains of B . thuringiensis, 4 strains of B . cereus and B . sphaericus have been detected respectively by multiple PCR . The results showed that 95.6% B . thuringiensis strains contain the B component of hblA gene, 91.1% and 93.3% of them contain bceT and entS genes sequences respectively . The enterotoxin productions in all strains have also been analysis using two commercial immunoassay kits(TECRA and RPLA) and it has proved that most of B . thuringiensis stains and the positive B . cereus strain can produce entero toxins during their growth . However, the two hblA sequence positive stains, DBT007 was negative when tested both by RPLA and Tecra, T24 001 was negative when assayed by Tecra and positive by RPLA . One hblA sequence negative strain Dmu39 was negative when tested by RPLA but positive by Tecra . No enterotoxin and enterotoxin gene could be detected in B . thuringiensis DBT248 and the B . sphaericus strains . The results suggest that the potential risk of using B . thuringiensis, as biopesticide needs to be further evaluated.

Wei Sheng Wu Xue Bao, 2000 Jun, 40(3), 264 - 9
{A new resolution vector with cry1Ac10 gene based on Bacillus thuringiensis transposon Tn4430}; Wu L et al.; A new resolution vector with cry1Ac10 gene based on TnpI-mediated site-specific recombination system of Bacillus thuringiensis(Bt) transposon Tn4430 was developed . The gene cry1Ac10, encoding a protoxin against plutella xylostella larvae, and the gene ori1030, from a plasmid of wide type Bacillus thuringiensis, were inserted into two copy sets of RES sites, named pBMB801 . When pBMB801 was introduced into crystal negative Bt host BMB171, antibiotic resistance genes and other non-Bt DNA can be selectively eliminated . This recombinant plasmid was found very stable without antibiotic selection . The resulting strain only contained Bt DNA and is free of antibiotic resistance genes . This strategy should facilitate regulatory approval for its development as a commercial biopesticide.

Wei Sheng Wu Xue Bao, 2000 Jun, 40(3), 252 - 6
{Infection of white spot baculo-like virus(WSBV) in Penaeus chinensis: evidence from electron microscopy and DNA hybridization}; Xu H et al.; Severe epizootic causing high mortalities in cultivated Penaeus chinensis has occurred every year in mainland China since 1993 with great economic loss . Naturally diseased P . chinensis sampled in Qingdao shrimp farming regions between 1993 and 1998 were examined by electron microscopy . A similar non-occluded nuclear bacilliform virus like the white spot baculo-like virus (WSBV) reported in Taiwan was mostly found in the gill, cuticular epidermis under the exoskeleton, stomach, lymphoid organ and other tissues of ectodermal and mesodermal origin from these diseased samples . The viron was enveloped with a mean size of (125 +/- 7.6) nm x (345 +/- 16) nm . Sometimes, a nipple-like appendage could be found protruding from one extremity of the viron . Envelopes were all lost after purification from cesium chloride gradient ultracentrifugation . The cylindrical nucleocapsid in negatively staining preparations was (80 +/- 13) nm x (380 +/- 24) nm with 13-16 stripes perpendicular to the longitudinal axis of the nucleocapsid . Based on its pathogenic and morphological characteristics, this virus should be related to WSBV . A pair of primers created from information of WSBV genome DNA SalI fragment produced a 355 bp band using the WSBV isolate from P . chinensis in mainland China, as the DNA template . The specific PCR product was cloned, sequenced and labeled with digoxigenin (DIG) DNA labeling kit (Boehringer Mannherm) . The sequence of the probe is identical to that of WSBV . All of the sampled diseased shrimps between 1993 and 1998 reacted with the DIG-labeled probe by dot-blot hybridization and no hybridization was observed using DNA from health shrimp as templates . These results based on electron microscopy and DNA hybridization indicated that the shrimp epizootic occurred in mainland China was related to the WSBV infection previously occurred in Taiwan early 1992.

Wei Sheng Wu Xue Bao, 2000 Aug, 40(4), 425 - 9
{Studies on biosorption of Au3+ by Bacillus megaterium}; Liu Y et al.; Some characteristics of Au3+ biosorption and bioreduction by Bacillus megaterium D01 biomass were reported . The optimum pH value of Au3+ biosorption by strain D01 was 3.0 . The biosorption was a rapid process, reaching 95% of the maximal capacity within the first 5 min of contact . Temperature did not affect the biosorption . The biosorptive efficiency(99.1%) and capacity (302.0 mg Au3+/g dry weight biomass) were achieved under the conditions of pH 3.0 and 30 degrees C, both concentration with an initial Au3+/biomass radio of 305 mg/g dry weight for 30 min of the contact . The D01 biomass could reduce Au3+ in the solution to Au0, Au0 on the cells surface and in the contact solution could be crystallized to different forms gold crystals, Au3+ impregnated on SiO2 and alpha-Fe2O3 could be reduced to Au0 by D01 biomass . The results of electrochemical reaction indicated that the strain D01 exhibited a high selectivity for Au3+.

Wei Sheng Wu Xue Bao, 2000 Aug, 40(4), 372 - 8
{Identification of cry-type genes of 31 Bacillus thuringiensis isolates and analysis of their expression product}; Zhang J et al.; In this study, cry-type gene of 31 isolates of Bt had been identified using PCR-RFLP identification system, and SDS-PAGE analysis and bioassay had been performed . 25 strains containing cry1 type gene express 130-150 kD protein and 16 of them contain cry1I gene, which codes 80 kD protein and is larvicidal to both lepidopteran and coleopteran species . 15 strains contain both cry1 and cry2 type gene; 10 strains contain unknown cry-type gene; 6 strains don't contain seven identified cry-type gene . Result of bioassay shows that the strains containing both cry1 and cry2 gene have high toxicity to lepidopteran order, 7 of them are strongly insecticidal to Pristiphora beijingensis and Lymantria dispar larvae; the strains containing cry1Ab + 1Ac + 2A or cry1Aa + 1Ac + 2A have high activity to Heliothis armigera; no cry gene, no activity . It is indicated that identification of cry-type gene and its analysis of SDS-PAGE protein can predict insecticidal activity of Bt isolates, and it is no pertinency between cry-type gene and serotype of Bt strain.

Wei Sheng Wu Xue Bao, 2000 Aug, 40(4), 365 - 71
{Characterization of the insecticidal crystal protein genes of Bacillus thuringiensis YBT-1520}; Sun M et al.; The insecticidal crystal protein genes of highly toxic Bacillus thuringiensis YBT-1520 were analyzed . Southern hybridization and PCR analysis indicated that this strain bears cry1Aa, cry1Ab, cry1Ac and cry2 genes . The copy number of three cry1A genes is different and cry1Ac gene (here named cry218) is the highest . The restriction location of three genes is different from that in typical strains, such as HD-1 and HD-73 strains . 4190 bp nucleotide sequence of cry218 gene was determined and this gene was named cry1Ac10 in the recently published gene nomenclature.

Wei Sheng Wu Xue Bao, 2000 Apr, 40(2), 139 - 42
{Expression and insecticidal characteristics of cry3A in Lepidoptera-specific Bacillus thuringiensis}; Yue C et al.; The cry3A gene coding Coleoptera-specific protein was transformed into a wild strain YBT803-1 by electroporation, A transformant BMBY-001 was obtained . SDS-PAGE analysis demonstrated that cry3A could express 64 kD protein in BMBY-001 . However, the expression of cry1Ab and cry1Ac were effected . Bioassay showed that BMBY-001 was not only highly toxic to Phyllodecta vulgatissima larva(LC50 0.413 microL/mL) but also toxic to Plutella xylostella(LC50 3.319 microL/mL).

Wei Sheng Wu Xue Bao, 1998 Dec, 38(6), 480 - 3
{Isolation and identification of N2-fixing strains of Bacillus in rice rhizosphere of the Yangtze River Valley}; Xie G et al.; Rice rhizosphere soil samples were colected from 10 sites of 7 provinces in the Yangzi River Valley, and from the soil samples 16 endospore-forming strains with ARA (Acetylene Reduction Activity) were isolated, the nitrogen fixing ability was tested by the method of 15N tracer and the atom 15N% excess are ranged from 0.0297% to 0.4714% . The strains were identified as Bacillus licheniformis, B . subtilis, B . azotoformans, B . cereus, B . pumilus, B . brevis, B . megaterium, B . firmu.

Wei Sheng Wu Xue Bao, 1998 Dec, 38(6), 422 - 7
{Ultrastructural differences of RMV and TMV infected Nicotiana tabacum mesophyll cells for distinguishing virus strains}; Xu J et al.; Nine strains of ribgrass mosaic virus (RMV) and tobacco mosaic virus(TMV) from cruciferous and solanaceous plants in different geographic regions of China were used to inoculate 4-leave-old plants of Nicotiana tobacum for observation of ultrastructually pathological changes due to infection of different strains . The samples of the mesophyll cells of the plants 15 d after inoculation were prepared in a routine and examined using transmission electronic microscopy . Characteristic virion arrangement, inclusion forms, and cell structural aberrations were found different among the various strains . The virions were accumulated in cluster, arranged in ring scattered, irregularly congregated with membrane, and aggregated in angle layer or parallel layer in cytoplasm and vacuoles whereas they were not found in organelles including nuclei, chloroplasts, and mitochondria . The inclusions with bacilliform, club-shaped or granulous X-tubule varied in number and form among the viral strains investigated . The abnormality of cell wall including thickening, collapsing and distortions were also observed among the cells infected by the strains . Other distinguishable changes attributed to the strains included the number, size, shape, and clumping pattern of chloroplasts, and of starch grains and osmiphilic globules chroloplasts of mesophyll cells infected only by two of the 9 strains examined . Additionally, the increased numbers of mitochondria and endoplasmic reticulum were observed in the cells infected by some of the strains . The results indicate that the ultrastructural changes due to viral infection may be useful for distinguishing various RMV and TMV strains.

Wei Sheng Wu Xue Bao, 2000 Feb, 40(1), 85 - 90
{Screening of acrystalliferous mutants from Bacillus thuringiensis and their transformation properties}; Li L et al.; After elevating growth temperature of YBT-1463 and other 8 Bacillus thuringiensis parental strains to 42 degrees C, 9 acrystalliferous(Cry-) mutants were obtained, and then a series of partial plasmid-cured mutants were further obtained from 3 Cry- mutants BMB151, BMB154 and BMB160 by elevating temperature to 44 degrees C . One of which, BMB170, was further treated by elevating temperature to 46 degrees C and adding 0.05% SDS as the plasmid-curing agent . The resulting mutant, which was designated BMB171 and had no resident plasmid, was finally obtained . The studies on the transformation efficiency and stability of Cry- mutants showed that the transformation frequencies, as well as stability of several exogenous plasmids(pHT3101, pBMB1736, etc.) introduced into Cry- mutants by electroporation, were correlated with the numbers of resident plasmids bearing by the relevant recipients, the patterns of plasmid replicons and the sizes of those exogenous plasmids . Meanwhile, a much higher electro-transformation frequencies of Cry- mutants than the parental strain YBT-1463 was found, and the highest of the transformation frequency, in BMB 171, could reach up to 10(7) transformants/microgram DNA, and the stability of those exogenous plasmids in BMB171 were also higher than in other Cry- mutants and YBT-1463 strain.

Wei Sheng Wu Xue Bao, 2000 Feb, 40(1), 62 - 8
{Study on the production of beta-mannanase by Bacillus M50}; Chen Y et al.; Bacillus sp . M50 produced beta-mannanase efficiently in the medium composed of 4% konjac powder, 1.0% (NH4)2SO4 and 0.35% Na2CO3 . Fermentation of beta-mannanase by Bacillus sp . M50 has been carried out in 100 L tank . The temperature, ventitation quantity and agitation speed were 30-32 degrees C, 1:0.75 vvm and 220 r/min respectively . the fermentation cycle was 28 h and the highest beta-mannanase activity was 330 u/mL . The optimal temperature and pH for beta-mannanase reaction were 50 degrees C and 6.0 respectively . It was stable at pH 4.0-7.0 and below 50 degrees C . The activity of the enzyme was strongly inhibited by Fe3+, Al3+, EDTA, Hg2+ and slightly stimulated by Ba2+, Mn2+ . Treatment of ramie-fibers with crude enzyme preparations from Bacillus sp . M50 showed a good degradation for hemicellulose of ramie-fibers residues.

Wei Sheng Wu Xue Bao, 2000 Oct, 40(5), 535 - 9
{Studies on biosorption of Pd2+ by bacteria}; Liu Y et al.; The strain R08, showed relatively strong ability of adsorbing Pd2+, was screened from different source of bacterial strains . The strain R08 was identified as Bacillus licheniformis . The optimum pH value of Pd2+ biosorption by R08 dead biomass is 3.5 . The biosorption is a rapid and non-temperature dependent process . The biomass and Pd2+ initial concentration in solution affected the biosorption process . The bosorptive capacity could reach 224.8 mg/g under the conditions of Pd2+ 200 mg/L, 0.4 g biomass/L, pH3.5 and 30 degrees C for 45 min . TEM observation showed that R08 biomass is able to adsorb and reduce Pd2+ ions to Pd0 particles . IR spectrum analysis indicated that -COO- and HPO(4)2- groups on the cell walls may involve in process of adsorbing Pd2+.

Eur J Immunol, 2003 Feb, 33(2), 306 - 13
The interaction between NK cells and dendritic cells in bacterial infections results in rapid induction of NK cell activation and in the lysis of uninfected dendritic cells; Ferlazzo G et al.; NK and DC reciprocal interactions have only recently been investigated . In this study, we focused on the interplay between NK cells and DC in two models of bacterial infection . Immature monocyte-derived DC were cultured in the presence of live Escherichia coli or bacillus Calmette-Guerin . Upon exposure to either extracellular or intracellular bacteria, DC underwent maturation as assessed by the increased levels of expression of CD80,CD86, and HLA molecules and the de novo expression of CD83 and CCR7 . Significant amounts of TNF-alpha and IL-12 were released by DC upon infection, whereas IL-2 and IL-15 were barely detectable in culture supernatants . Both infected and uninfected DC were capable of inducing in fresh autologous NK cells the expression of CD69 and HLA-DR and of inducing cell proliferation . Remarkably, however, infected DC were much stronger inducers of NK cell activation and proliferation than uninfected DC . Thus, after just 24 h of NK/DC coculture, only those NK cells that had been exposed to bacteria-infected DC had acquired the ability to lyse autologous immature DC . In addition, infected DC were more resistant to NK-mediated lysis as a consequence of the up-regulation of HLA class I molecule expression on their surface . This study suggests a regulatory circuit involving NK cells and DC in which DC-induced NK cell activation is effectively enhanced by the presence of pathogens . Activated NK cells, by limiting the supply of immature DC, may then exert a control on subsequent innate and adaptive immune responses.

Lancet, 2003 Jan 18, 361(9353), 239 - 46
Whipple's disease; Marth T et al.; Whipple's disease, or intestinal lipodystrophy, is a systemic infectious disorder affecting mostly middle-aged white men . Patients present with weight loss, arthralgia, diarrhoea, and abdominal pain . The disease is commonly diagnosed by small-bowel biopsy; the appearance of the sample is characterised by inclusions in the lamina propria staining with periodic-acid-Schiff, which represent the causative bacteria . Tropheryma whipplei has been classified as an actinomycete and has been propagated in vitro, which allows the possibility of improving diagnostic strategies, for example through antibody-based detection of the bacillus on duodenal tissue or in circulating monocytes . Cell-mediated immunity in active and inactive Whipple's disease has subtle defects that might predispose some individuals to symptomatic infection with this bacillus, which probably occurs ubiquitously . Although most patients respond well to empirical antibiotic treatment, some with relapsing disease have a poor outlook . The recent findings and concerted research might allow development of new strategies for diagnosis, treatment, and monitoring of patients with Whipple's disease.

J Mol Biol, 2003 Feb 7, 326(1), 177 - 88
Three-dimensional structure and substrate binding of Bacillus stearothermophilus neopullulanase; Hondoh H et al.; Crystal structures of Bacillus stearothermophilus TRS40 neopullulanase and its complexes with panose, maltotetraose and isopanose were determined at resolutions of 1.9, 2.4, 2.8 and 3.2A, respectively . Since the latter two carbohydrates are substrates of this enzyme, a deactivated mutant at the catalytic residue Glu357-->Gln was used for complex crystallization . The structures were refined at accuracies with r.m.s . deviations of bond lengths and bond angles ranging from 0.005A to 0.008A and 1.3 degrees to 1.4 degrees, respectively . The active enzyme forms a dimer in the crystalline state and in solution . The monomer enzyme is composed of four domains, N, A, B and C, and has a (beta/alpha)(8)-barrel in domain A . The active site lies between domain A and domain N from the other monomer . The results show that dimer formation makes the active-site cleft narrower than those of ordinary alpha-amylases, which may contribute to the unique substrate specificity of this enzyme toward both alpha-1,4 and alpha-1,6-glucosidic linkages . This specificity may be influenced by the subsite structure . Only subsites -1 and -2 are commonly occupied by the product and substrates, suggesting that equivocal recognition occurs at the other subsites, which contributes to the wide substrate specificity of this enzyme.

Dermatol Online J . 2002 Oct;8(2):6.
Short course of clarithromycin in an immunocompetent patient with BCG-induced regional complications; Torres-Rojas JR et al.; Bacillus Calmette Guerin (BCG) has been used to vaccinate against tuberculosis since 1921 . Persistant skin and lymph node lesions yielding Mycobacterium bovis are a rare complication for which there is not standardized treatment . We report an 11 month old child with a progressive cutaneous nodule and lymphadenopathy after BCG vaccination . These rapidly cleared with administration of clarithromycin.

J Am Mosq Control Assoc, 2002 Dec, 18(4), 341 - 3
Laboratory evaluation of Vectobac as against Aedes aegypti in Monterrey, Nuevo León, Mexico; Ponce G G et al.; Intensive use of the organophosphate insecticide malathion against adults and temephos against larvae of Aedes aegypti in Mexico over the past 30 years has led to problems requiring the use of new larvicides . Toward this objective, Bacillus thuringiensis var . israelensis (Bti), a target-specific and environmentally safer control agent, was evaluated . Laboratory bioassays were done to determine the susceptibility of 2nd- and 3rd-stage larvae of Ae . aegypti to Vectobac 12 AS (aqueous suspension, 600 ITU/mg) . A median lethal concentration of 0.0104 ppm and a 95% lethal concentration of 0.18 ppm were determined after 24 h of exposure to the agent . The values obtained were adjusted for field application and were further tested in the field by the State of Nuevo Leon, Mexico Vector Control Program . Suspensions of Bti were poured into pipe-water trucks and transferred to domestic 200-gal metal water drums . Larval populations were reduced during a 2-week study period . However, residents complained about a fine dusty film on the water surface . Nevertheless, these results are promising for future Bti field applications.

Hua Xi Yi Ke Da Xue Xue Bao, 2000 Sep, 31(3), 285 - 8
{Enhancement of BCG-induced hBD-1 mRNA expression in human pulmonary gland epithelial cells}; Feng Y et al.; Using reverse-transcription PCR (RT-PCR) and Northern blot analysis, the present authors found an enhanced expression of the human beta-defensin 1 (hBD-1) gene in the SPC-A-1 cells challenged with heat-killed M . Bovis Bacille Calmette-Guerin (BCG) . The enhanced expression of hBD-1 mRNA was dose- and time-dependent . This result indicated a significant progress in the work of developing a new antimicrobial strategy that enhances mucosa antibiotic peptide expression for the prevention and treatment of mucosal infections.

Klin Padiatr, 2003 Jan-Feb, 215(1), 24 - 9
{Early and late complications after implantation of central venous catheters}; Herold A et al.; The use of implantable central venous catheters by the puncture-technique of Nagy is a safe method performed by trained surgeons . Due to the high mobility and then reduction of painful blood samples this method contributes favourably to the improvement of the quality of life of children with chronic diseases . The analysis of 140 catheters implanted in the Department of Pediatric Surgery of the University of Leipzig between 1995 and 2000 showed 11 cases with early complications . As the most frequent late complications were infection and thrombosis in 51 children . Neutropenia is a particular risk factor during polychemotherapy of malignant tumors . Staphylococcus epidermidis was the most frequently isolated bacillus . Catheter associated infections are only partly treatable by antibiotic therapy . In 29 of 44 cases the explantation of the catheter was necessary . Only a strict hygienic regimen would minimize the risk of infection

J Ind Microbiol Biotechnol, 2003 Jan, 30(1), 47 - 51 Epub 2003 Jan 03.
Immobilisation of cyclodextrin glucanotransferase from Bacillus circulans ATCC 21783 on purified seasand; Iyer JL et al.; Cyclodextrin glucanotransferase (CGTase) from Bacillus circulans (ATCC 21783) was immobilised on a silica-based support: purified seasand . Although adsorption of 98% was achieved, considerable desorption was encountered . This problem was minimised by crosslinking the adsorbed enzyme with glutaraldehyde . The immobilised enzyme after crosslinking could be used repeatedly for cyclodextrin (CD) production in a batch process . The activity retention was 80% at the end of the eighth cycle . The immobilised enzyme showed a shift in the pH optimum towards the alkaline side and also an improvement in the pH stability compared to the free enzyme . It catalysed the formation of beta-CD as a major product . A significant amount of alpha-CD production was also observed on prolonged incubation.

AIDS Patient Care STDS, 2002 Dec, 16(12), 573 - 7
AIDS presenting with cutaneous Kaposi's sarcoma and bacillary angiomatosis in the bone marrow mimicking Kaposi's sarcoma; Rosales CM et al.; Kaposi's sarcoma (KS) and bacillary angiomatosis (BA) may be histologically similar . A precise diagnosis is required because of the different management of these diseases . KS or BA involving bone marrow is rare in patients with and without acquired immune deficiency syndrome (AIDS) . We report the case of a 40-year-old human immunodeficiency virus (HIV)-positive homosexual male who presented with small KS lesions in the skin and BA in the bone marrow that histologically were similar . Laboratory evaluation revealed anemia and thrombocytopenia; CD4 count was 103/mm3, and the viral load was 750,000 HIV-1 mRNA copies per milliliter in plasma . Bartonella henselae, the etiologic agent of BA, was isolated from a blood culture . DNA sequences of human herpesvirus-8 (HHV-8), the putative etiologic agent of KS, were identified by polymerase chain reaction (PCR) in skin and bone marrow specimens, but antibody anti-HHV-8-encoded protein ORF73, localized signals only in the skin-KS lesion . The patient received clarithromycin and cefotetan for the BA, and antiretroviral therapy for the HIV infection . The skin lesions gradually regressed, the HIV-1 mRNA copy number decreased to less than 400 per milliliter and the CD4 lymphocyte count increased to 665/mm3 . In conclusion, vascular lesions of BA and KS may be clinically and histologically similar, both may be associated with advanced AIDS, and an accurate diagnosis is required because of their different management.

J Am Mosq Control Assoc, 2002 Dec, 18(4), 352 - 8
Evaluation of polymer-based granular formulations of Bacillus thuringiensis israelensis against larval Aedes aegypti in the laboratory; Maldonado Blanco MG et al.; A strategy to increase residual activity of Bacillus thuringiensis serovar . israelensis (Bti) extract through slow-release formulations and protection from solar radiation was studied . The median lethal concentration (LC50) and 90% lethal concentration (LC90) levels of laboratory-reared early 4th-stage larval Aedes aegypti after exposure to Bti extract were determined . Formulations with 4 polymers and 1 solar protectant were prepared, and their shelf lives were evaluated for year-long storage at 20-35 degrees C and 50-80% relative humidity . Also, the effect of ultraviolet light on unformulated (extract) and formulated Bti larvicidal activity persistence was determined . Laboratory bioassays were conducted with larval Ae . aegypti introduced into treated and control containers at 1, 2, 7, 14, 21, 28, 35, and 42 days after treatment, and larval mortalities were checked 24 h after introduction . Probit analysis of Bti extract showed LC50 and LC90 values of 0.016 and 0.051 mg/liter, respectively . The polymer-based Bti formulations showed no significant loss of insecticidal activity after 8 months of storage . Ultraviolet irradiation reduced activity of unformulated Bti extract after different exposure times, up to 40-46%, whereas Bti formulated with gelatin or acacia gum showed lower variation in larvicidal activity than formulations with sodium alginate and paraffin for protecting the activity of Bti toxin . Residual activity against 4th-stage Ae . aegypti in the laboratory for the formulation containing acacia gum at 10% (w/w) was 80% mortality at 14 days after treatment, whereas the Bti formulation containing gelatin (10%, w/w) caused 65% mortality . In addition, Bti formulations made with paraffin at 5% (w/w) sustained up to 60% mortality for 21 days . Unformulated Bti showed only 2.6% mortality, and a commercial preparation maintained 37% mortality, both at 14 days after treatment.

J Am Mosq Control Assoc, 2002 Dec, 18(4), 344 - 7
Operational studies with Valent VectoLex WDG, Bacillus sphaericus, in three Florida Mosquito Control Districts; Floore T et al.; VectoLex WDG, Bacillus sphaericus (Valent BioSciences Corporation), was evaluated in 3 Florida Mosquito Control Districts under operational conditions . Application rates were 0.5 and 1.0 lb/acre and an untreated control in each district . Study sites included woodland ponds, residential areas such as catch basins, and commercial business retention ponds . Pretreatment and posttreatment assessments were made with a standard dipper at designated dipping stations . The most prevalent mosquitoes included Culex quinquefasciatus, Culex nigripalpus, Culex erraticus, Ochlerotatus infirmatus, Anopheles crucians, Anopheles quadrimaculatus, and Psorophora spp . VectoLex WDG was effective for 24-35 days posttreatment with one application.

J Biol Chem, 2003 Mar 28, 278(13), 11546 - 53 Epub 2003 Jan 21.
Kinetic stabilization of Bacillus licheniformis alpha-amylase through introduction of hydrophobic residues at the surface; Machius M et al.; It is generally assumed that in proteins hydrophobic residues are not favorable at solvent-exposed sites, and that amino acid substitutions on the surface have little effect on protein thermostability . Contrary to these assumptions, we have identified hyperthermostable variants of Bacillus licheniformis alpha-amylase (BLA) that result from the incorporation of hydrophobic residues at the surface . Under highly destabilizing conditions, a variant combining five stabilizing mutations unfolds 32 times more slowly and at a temperature 13 degrees C higher than the wild-type . Crystal structure analysis at 1.7 A resolution suggests that stabilization is achieved through (a) extension of the concept of increased hydrophobic packing, usually applied to cavities, to surface indentations, (b) introduction of favorable aromatic-aromatic interactions on the surface, (c) specific stabilization of intrinsic metal binding sites, and (d) stabilization of a beta-sheet by introducing a residue with high beta-sheet forming propensity . All mutated residues are involved in forming complex, cooperative interaction networks that extend from the interior of the protein to its surface and which may therefore constitute "weak points" where BLA unfolding is initiated . This might explain the unexpectedly large effect induced by some of the substitutions on the kinetic stability of BLA . Our study shows that substantial protein stabilization can be achieved by stabilizing surface positions that participate in underlying cooperatively formed substructures . At such positions, even the apparently thermodynamically unfavorable introduction of hydrophobic residues should be explored.

J Econ Entomol, 2002 Dec, 95(6), 1205 - 15
Evaluation of preventive treatments in low-density gypsy moth populations using pheromone traps; Sharov AA et al.; Pheromone traps can be used for evaluating the success of treatments that are applied to either eradicate or delay the growth of isolated low-density populations of the gypsy moth, Lymantria dispar (L.) . We developed an index of treatment success, T, that measures the reduction in moth counts in the block treated adjusted by the change in moth counts in the reference area around it . This index was used to analyze the effectiveness of treatments that were conducted as part of the USDA Forest Service Slow-the-Spread of the gypsy moth project from 1993 to 2001 . Out of 556 treatments that were applied during this period, 266 (188,064 ha) were selected for the analysis based on several criteria . They included 173 blocks treated with Bacillus thuringiensis (Berliner) variety kurstaki and 93 blocks treated with racemic disparlure . Analysis using general linear models indicated that disparlure treatments were significantly more effective than B . thuringiensis treatments in reducing moth captures . The frequency of repeated treatments in the same area was higher after B . thuringiensis than after disparlure applications . Treatments were more successful if the pretreatment moth counts outside of the block treated were low compared with moth counts inside the block.

Prostate, 2003 Mar 1, 54(4), 322 - 30
Characteristics of human prostasomes isolated from three different sources; Carlsson L et al.; BACKGROUND: Prostasomes are organelles secreted by human prostate epithelial cells and appear freely in semen . Most prostasomal functional and biochemical characteristics have been settled on basis of seminal prostasomes (SP) . METHODS: Three types of prostasomes, viz., from seminal plasma, prostate gland (native), and prostate cancer bone metastasis, were isolated and compared for their biochemical and functional characteristics . RESULTS: Flow cytometry showed that all three types of prostasomes had a similar size distribution and electrophoresis revealed close similarities in banding pattern . Also, a high cholesterol/phospholipid ratio and some marker enzymes were distinctive common features . The unusual chromogranin B (CgB)/chromogranin A (CgA) ratio with CgB in abundance over CgA, previously found only in SP, was also valid for the native (NP) and metastasis-derived prostasomes (MP) . The latter prostasome type contained a 10-fold higher amount of CgA compared to the others . SP and MP showed the same promotive effect on sperm forward motility, whereas NP did not . All three prostasome types exhibited an antibacterial activity against Bacillus megaterium . CONCLUSIONS: Since the three prostasome types were isolated from different environments, our data support the view that the common characteristics were prostasome-genuine .

J Immunol, 2003 Feb 1, 170(3), 1392 - 8
The shift of Th1 to Th2 immunodominance associated with the chronicity of Mycobacterium bovis bacille Calmette-Guérin infection does not affect the memory response; Jiao X et al.; In the present study we investigated the shaping and evolution of the immunodominance of the T cell response during a chronic mycobacterial infection . Using a recombinant bacille Calmette-Guerin expressing a reporter Ag, the Escherichia coli MalE protein, we analyzed the peptide specificity and the cytokine profile of the T cell response to the reporter Ag by ELISPOT . During the early steps of infection, the T cell response was focused on two dominant MalE epitopes and was characterized by a pure IFN-gamma response . Then, in the course of infection the initial IFN-gamma response to these two epitopes shifted to a mixed IFN-gamma/IL-4 response . At the same time, the peptide specificity of the T cell response was broadened to two additional MalE epitopes characterized by a unique IL-4 response resulting in the establishment of a dominant IL-4 response to the MalE protein at 16 wk postinfection . However, this phenomenon did not impair the outcome of a predominant IFN-gamma response upon subsequent MalE recall in vivo performed in the presence of CFA, a Th1-driving adjuvant . These results indicate that the Th2 nature of the immune response established during a chronic infection, which most likely reflects regulatory mechanisms to allow the return to T cell homeostasis, does not shape the Th1/Th2 nature of the memory response.

Appl Microbiol Biotechnol, 2003 Jan, 60(5), 588 - 93 Epub 2002 Nov 20.
Characteristics of a bioflocculant produced by Bacillus mucilaginosus and its use in starch wastewater treatment; Deng SB et al.; A bioflocculant, MBFA9, was produced from a strain of bioflocculant-producing bacteria isolated from a soil sample and identified as Bacillus mucilaginosus . MBFA9 had a good flocculating capability and could achieve a flocculating rate of 99.6% for kaolin suspension at a dosage of only 0.1 ml/l . The major component of MBFA9 was found to be polysaccharide composed mainly of uronic acid (19.1%), neutral sugar (47.4%) and amino sugar (2.7%) . Infrared spectrum analysis showed the presence of carboxyl and hydroxyl groups in the bioflocculant . MBFA9 is nontoxic and can be used in food industries for suspended solids (SS) recovery . When applied to starch wastewater treatment, MBFA9 greatly accelerated the formation of flocs and the settling of organic particles in the presence of Ca(2+) salt . After 5 min of settling, the removal rate of SS and chemical oxygen demand were up to 85.5% and 68.5%, respectively, which is better than traditional chemical flocculants.

Trop Med Int Health, 2003 Jan, 8(1), 37 - 47
Efficacy and efficiency of new Bacillus thuringiensis var israelensis and Bacillus sphaericus formulations against Afrotropical anophelines in Western Kenya; Fillinger U et al.; We evaluated the efficacy of new water-dispersible granular (WDG) formulations of Bacillus thuringienis var . israelensis (Bti; VectoBac) and B . sphaericus (Bs; VectoLex), Valent BioScience Corp., Illinois, USA) for the control of larval Anopheles gambiae sensu lato Giles mosquitoes in a malaria-endemic area around Lake Victoria, Western Kenya . WDG and powder formulations were compared in laboratory bioassays and followed by efficiency and residual effect assessments of both WDG formulations in open field experiments . LC50 and LC95 values for the Bti/Bs strains and their formulations show high susceptibility of A . gambiae sensu stricto under laboratory conditions . The larvae proved more susceptible to Bs than to Bti and the WDG formulations were slightly superior to the powder formulations . High efficiency was also shown in the open field trials, and a minimum dosage of 200 g/ha Bti WDG, representing the LC95 of the laboratory tests, was sufficient to fully suppress emergence of mosquitoes when applied at weekly intervals . Bti WDG did not show a residual effect, irrespective of the concentration applied . The Bs WDG formulation, however, showed significant larval reductions up to 11 days post-treatment at application doses of either 1 or 5 kg/ha . We conclude that the main malaria vector in our study area is highly susceptible to these microbial control agents . Minimum effective dosages to achieve elimination of the larval population in a given habitat are extremely low and environmental impact is negligible . Microbial products for larval control have therefore great potential within Integrated Vector Management programmes and may augment control efforts against adult vector stages, such as the use of insecticide-treated bednets, in many parts of Africa.

Immunol Cell Biol, 2003 Feb, 81(1), 79 - 85
Aspects of cancer immunotherapy; McKenzie IF et al.; Cancer immunotherapy has traditionally undergone a 'revolution' every decade, from the use of Bacille Calmette-Guerin by scarification in the 1970s, to interleukin-2 therapies in the 1980s, and monoclonal antibody treatments in the early 1990s . Usually the early reports on the use of such agents were encouraging, but when more patients were studied in multiple centres, the initial promising results could not be confirmed . Now in a new century, we have more reagents and methods available than ever before - indeed, with such a plethora of reagents it is difficult to envisage them being fully and appropriately tested within the next decade, by which time there will be even more reagents to test . However, there have been three major advances which should lead to substantial progress in cancer immunotherapy: (1) the widespread use of genetic engineering, enabling identification of candidate vaccine proteins and manipulation of their sequences; (2) the production of antigens, antibodies and cytokines in large amounts by recombinant technologies, and (3) an understanding of the mode of presentation of peptides by major histocompatibility complex Class I and Class II molecules and their recognition by T cells . Despite these advances, there are major problems facing cancer immunotherapy, such as the ability of tumours to mutate and evade the immune system and the difficulty of precisely defining the interactions of effector cells in mediating 'rejection' or destruction of a tumour . There are clearly immunological similarities with diseases such as malaria and schistosomiasis, where the invading foreign organisms can use a variety of strategies to resist an elicited immune response . The failure to find a suitable vaccine for these diseases must lead to some pessimism for the development of immunotherapy for an autologous tumour . However, there are promising studies now in progress which should give an indication of the most important directions to follow . This review provides a commentary on aspects of cancer immunotherapy and in particular will deal with: (1) the selection of antigens as vaccine components; (2) the modes of presentation of antigens, particularly by major histocompatibility complex Class I molecules; and (3) new modes of delivery of vaccine immunogens.

J Appl Microbiol, 2003, 94(2), 301 - 11
Initial characterization of new bacteria degrading high-molecular weight polycyclic aromatic hydrocarbons isolated from a 2-year enrichment in a two-liquid-phase culture system; Gauthier E et al.; AIMS: To characterize some polycyclic aromatic hydrocarbons (PAH)-degrading microorganisms isolated from an enriched consortium degrading high molecular weight (HMW) PAHs in a two-liquid-phase (TLP) soil slurry bioreactor, and to determine the effect of low molecular weight (LMW) PAH on their growth and HMW PAH-degrading activity . METHODS AND RESULTS: Several microorganisms were isolated from a HMW-PAH (pyrene, chrysene, benzo{a}pyrene and perylene) degrading consortium enriched in TLP cultures using silicone oil as the organic phase . From 16S rRNA analysis, four isolates were identified as Mycobacterium gilvum B1 (99% identity),Bacillus pumilus B44 (99% identity), Microbacterium esteraromaticum B21 (98% identity), and to the genus Porphyrobacter B51 (96% identity) . The two latter isolates have not previously been associated with PAH degradation . Isolate B51 grew strongly in the interfacial fraction in the presence of naphthalene vapours and phenanthrene compared with cultures without LMW PAHs . Benzo{a}pyrene was degraded in cultures containing a HMW PAH mixture but pyrene had no effect on its degradation . The growth of isolates B1 and B21 was improved in the aqueous phase than in the interfacial fraction for cultures with naphthalene vapours . Pyrene was required for benzo{a}pyrene degradation by isolate B1 . For isolate B21, pyrene and chrysene were degraded only in cultures without naphthalene vapours . CONCLUSION: Consortium enriched in a TLP culture is composed of microorganisms with different abilities to grow at the interface or in the aqueous phase according to the culture conditions and the PAH that are present . Naphthalene vapours increased the growth of the microorganisms in TLP cultures but did not stimulate the HMW PAH degradation . SIGNIFICANCE AND IMPACT OF THE STUDY: New HMW PAH-degrading microorganisms and a better understanding of the mechanisms involved in HMW PAH degradation in TLP cultures.

Biochemistry, 2003 Jan 28, 42(3), 739 - 45
Structure and dynamics of reduced Bacillus pasteurii cytochrome c: oxidation state dependent properties and implications for electron transfer processes; Bartalesi I et al.; The solution structure of reduced Bacillus pasteurii cytochrome c, which has only 71 amino acids, has been determined by NMR to an RMSD of 0.46 +/- 0.08 A for all backbone atoms and 0.79 +/- 0.08 A for all heavy atoms and refined through restrained energy minimization . The target function out of 1645 constraints is 0.52 +/- 0.11 A(2), and the penalty function is 66 +/- 12 kJ mol(-)(1) . The structure appears very similar to that in the oxidized state, only Trp87 and the propionates showing significant differences . The mobility was investigated through (15)N R(1) and R(2) relaxation rates, (15)N-(1)H NOE, and (1)H/(2)H exchange . It is found that the oxidized form is generally more mobile than the reduced one . By comparing the redox-state dependence of the structural/dynamic properties of Fe-S proteins, cytochrome c, and blue copper proteins, hints are provided for a better comprehension of the electron transfer processes.

World J Gastroenterol, 2003 Feb, 9(2), 320 - 3
Effect of leflunomide on immunological liver injury in mice; Yao HW et al.; AIM: To study the effect of leflunomide on immunological liver injury (ILI) in mice . METHODS: ILI was induced by tail vein injection of 2.5 mg Bacillus Calmette-Guerin (BCG), and 10 d later with 10 microg lipopolysaccharide (LPS) in 0.2 mL saline (BCG+LPS) . The alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO) level in plasma and molondiadehyde (MDA), glutathione peroxidase (GSHpx) in liver homogenate were assayed by spectroscopy . The serum content of tumor necrosis factors-alpha (TNF-alpha) was determined by ELISA . Interleukin-1 (IL-1), interleukin-2 (IL-2) and Concanavalin A (ConA)-induced splenocyte proliferation response were determined by methods of (3)H-infiltrated cell proliferation . RESULTS: Leflunomide (4, 12, 36 mg.kg(-1)) was found to significantly decrease the serum transaminase (ALT, AST) activity and MDA content in liver homogenate, and improve reduced GSHpx level of liver homogenate . Leflunomide (4, 12, 36 mg.kg(-1)) significantly lowered TNF-alpha and NO level in serum, and IL-1 produced by intraperitoneal macrophages(PMphi) . Moreover, the decreased IL-2 production and ConA-induced splenocyte proliferation response were further inhibited . CONCLUSION: These findings suggested that leflunomide had significant protective action on ILI in mice.

Vaccine, 2003 Jan 30, 21(7-8), 667 - 70
Mycobacterium bovis BCG-based vaccines against tuberculosis: novel developments; Dietrich G et al.; Mycobacterium bovis Bacille Calmette-Guerin (BCG) is one of the most widely used vaccines . Modern techniques in genome manipulation allow the construction of recombinant (r)-BCG strains that can be employed as highly immunogenic vaccines against tuberculosis (TB) with an enhanced safety profile . In addition, the development of novel procedures to cultivate BCG will allow the large-scale production of future BCG-based vaccines.

Biotechnol Appl Biochem, 2003 Jun, 37(Pt 3), 295 - 9
Purification and properties of a transglutaminase produced by a Bacillus circulans strain isolated from the Amazon environment; de Barros Soares LH et al.; A new microbial transglutaminase (EC 2.3.2.13) from a Bacillus circulans strain isolated from the aquatic Amazonian environment was purified and characterized . Enzyme purification started with (NH(4))(2)SO(4) 'salting out' and proceeded with liquid chromatography on Q-Sepharose FF and octyl-Sepharose 4 FF . The purification factor was approx . 150-fold with a yield of 32% . The enzyme's molecular mass was estimated as 45000 Da on SDS/PAGE . The purified transglutaminase had an optimum temperature of 47 degrees C, the optimum pH of the reaction was 7 and it presented no calcium-dependent activity.

APMIS, 2002 Sep, 110(9), 625 - 9
Bacillarity at autopsy in pulmonary tuberculosis . Mycobacterium tuberculosis is often disseminated; Lillebaek T et al.; The aim of this investigation was to quantify dissemination of Mycobacterium tuberculosis infection in patients with pulmonary tuberculosis and to show the pattern of eradication during treatment . The study is based on 98 out of the 113 patients with pulmonary tuberculosis who died during their admission to hospital in the Municipality of Copenhagen from 1963 to 1971 . These patients had cultures for M . tuberculosis performed from different organs at autopsy: 78% treated <=100 days had dissemination of bacteria, cultured with decreasing frequency in the lungs, spleen, liver, and kidneys, respectively . In comparison, 23% treated >100 days had dissemination of bacteria, among which 50% occurred in patients with records of poor treatment compliance, 14% in patients with good treatment compliance . 81% of all patients had at least one chest x-ray judged to be without a miliary pattern . This study emphasizes that M . tuberculosis is often disseminated to organs other than the lungs in severe pulmonary tuberculosis . Eradication of bacteria in these organs can take several months . This observation adds to our understanding of the natural history of tuberculosis: M . tuberculosis is a resilient organism that can adapt to a wide variety of environmental conditions.

Chest, 2003 Jan, 123(1), 107 - 12
Serologic diagnosis of tuberculosis using a simple commercial multiantigen assay; Perkins MD et al.; SETTING: Seven primary health clinics and a pulmonary disease specialty clinic in Rio de Janeiro City, Brazil . OBJECTIVE: To evaluate a commercial immunochromatographic test kit (ICT Tuberculosis; AMRAD-ICT; Sidney, Australia) employing five recombinant Mycobacterium tuberculosis proteins for the detection of pulmonary tuberculosis (TB) . DESIGN: Serology test results were compared with duplicate sputum microscopy and culture in 277 patients with symptomatic pulmonary disease (243 with pulmonary TB and 34 with nontuberculous disease) . An additional 110 healthy control subjects were also tested . RESULTS: The serology test was simple and rapid to perform and detected 64.2% of smear-positive and 46.3% of smear-negative TB patients overall . HIV co-infection was present in 15.3% of TB patients, and serology was much less sensitive (overall 27.6%) in this small group, as was microscopy (13.8%) . Specificity of the serology test was 100% in healthy control subjects and 85.2% in the small number of control patients with pulmonary disease, including those with prior TB . Combined with microscopy, serology detected 72.8% of TB patients . CONCLUSION: Depending on the population studied, multiantigen serologic tests for TB may be as sensitive as microscopy, but detect a different and overlapping subset of patients . The use of multiple antigens in this kit increased test sensitivity without significant loss of specificity . Bacille Calmette-Guerin vaccination and tuberculin sensitivity did not affect serology results . Estimating specificity in clinical use will require testing a much larger cohort of symptomatic patients with nontuberculous disease . The TB diagnostic performance of this group of antigens in HIV co-infected individuals was poor.

Immunol Lett, 2003 Jan 22, 85(2), 207 - 11
A novel TB vaccine; strategies to combat a complex pathogen; Olsen AW et al.; Years of intense research aimed at developing a new and improved vaccine against tuberculosis (TB) are now moving from the laboratories and into the field . Many groups have provided strong evidence for the potential of novel vaccines using a variety of different strategies and the most promising are just beginning to enter human safety trials . As research has developed over the years, the complexity of the TB situation at the global level and its influence on vaccine development strategies has become more and more clear . From being mainly focused on the development of a vaccine that could be given at birth and provide lifelong protection, it is now clear that the current goal may need to be a vaccine to be given to a population where the majority is already sensitized either by previous infection, exposure to other mycobacteria or by Bacille Calmette-Guerin vaccination . With the increasing awareness of the different immune mechanisms operating under those circumstances, such a vaccine still represents a significant challenge for immunologists and microbiologists working in this field.

Cell, 2002 Dec 13, 111(6), 815 - 24
Crystal structures of the Bacillus stearothermophilus CCA-adding enzyme and its complexes with ATP or CTP; Li F et al.; CCA-adding enzymes polymerize CCA onto the 3' terminus of immature tRNAs without using a nucleic acid template . The 3.0 A resolution crystal structures of the CCA-adding enzyme from Bacillus stearothermophilus and its complexes with ATP or CTP reveal a seahorse-shaped subunit consisting of four domains: head, neck, body, and tail . The head is structurally homologous to the palm domain of DNA polymerase beta but has additional structural features and functions . The neck, body, and tail represent new protein folding motifs . The neck provides a specific template for the incoming ATP or CTP, whereas the body and tail may bind tRNA . Each subunit has one active site capable of switching its base specificity between ATP and CTP, an important component of the CCA-adding mechanism.

J Biotechnol, 2003 Feb 27, 101(1), 69 - 79
A thermostable alpha-arabinofuranosidase from xylanolytic Bacillus pumilus: purification and characterisation; Degrassi G et al.; Bacillus pumilus PS213 secretes an alpha-L-arabinofuranosidase (AF) when grown in the presence of arabinogalactan or oat meal . The enzyme has been purified to homogeneity and characterised . Its molecular mass, as determined by gel filtration, is 220 kDa, while sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a single band of approximately 60 kDa . According to the result of the mass spectrometry analysis showing a molecular mass of 56 kDa, the enzyme should be a homotetramer . The isoelectric point was found to be 5.2, the enzyme activity was optimal at 55 degrees C and pH 7.0 . The enzyme retained 80% of its activity after 2 h at 65 degrees C and lost 50% of activity at 75 degrees C after 135 min . The Michaelis constant K(m) and V(max) for p-nitrophenylarabinofuranoside at 37 degrees C were 1.7 mM and 52.9 U mg(-1), respectively . N-terminal sequence analysis and internal peptide fragments showed homology with glycosyl hydrolases of family 51.

Water Sci Technol, 2002, 46(11-12), 257 - 62
Characterization of an algae-lytic substance secreted by Bacillus cereus, an indigenous bacterial isolate from Lake Kasumigaura; Nakamura N et al.; In order to develop a practical biological strategy for water bloom-control in Lake Kasumigaura, indigenous algae-lytic bacteria targeting Microcystis spp . (the dominant cyanobacteria), was first carried out . Some basic lytic characteristics of the isolates concerning the biological and physico-chemical factors in actual water environments were then investigated . Fifteen algae-lytic bacteria were successfully isolated using the double layer method . The isolate showing the highest lytic activity was identified as Bacillus cereus based on the 16SrRNA sequence . Extracellular products of B . cereus were found to be responsible for algae-lytic activity . Algae-lytic assay tests using bacterial supernatants pre-treated under several conditions indicated that the majority of the effective algae-lytic substances were either hydrophilis or hydrophobics with a carbon number less than 18, and with molecular weight less than 2000 Da . The algae-lytic activity of the bacterial supernatant was found to be enhanced under alkaline conditions, but it was lost under acidic conditions . This pH specific characteristic is advantageous for application in water bloom environments where the pH is usually in the alkaline region.

Neurol Neurochir Pol, 2002 Sep-Oct, 36(5), 959 - 70
{Whipple's disease--a rare cause of neurological symptoms and disorders}; Slowik A et al.; Whipple's disease is a chronic infection caused by Gram-positive bacillus Thropheryma Whippelii picture with a wide range of clinical manifestation, not only systemic but also neurological . Seronegative arthritis or arthralgia may be the only presenting symptom, predating by years gastrointestinal, and also pulmonary, cardiac, renal and neurological manifestations . The diagnosis can be established based on the characteristic histopathological features found in the affected organ (foamy macrophages with a coarsely granular cytoplasm, which stains with PAS) and PCR of 16S ribosomal RNA of Tropheryma Whippelii . CNS involvement manifests with a broad range of neurological symptomatology: memory, consciousness, hypothalamic, psychiatric and behavioural disorders and other symptoms, which may mimic neurodegeneration, neuroinfection, stroke and tumour . In this review detailed neurological symptomatology, differential diagnosis and laboratory, neurophysiologic and radiologic findings are presented . Whipple's disease is potentially fatal but responds to antibiotic treatment . The current recommendations for treatment are discussed.

Clin Infect Dis, 2003 Jan 15, 36(2), 140 - 8 Epub 2003 Jan 03.
Spectrum of bacille Calmette-Guérin (BCG) infection after intravesical BCG immunotherapy; Gonzalez OY et al.; Intravesical instillation of bacille Calmette-Guerin (BCG) effectively treats transitional cell carcinoma of the bladder . Occasionally, BCG infection complicates such treatment . In some patients, infection appears early (within 3 months after instillation) and is characterized by generalized symptoms, with pneumonitis and hepatitis . Late-presentation disease occurs >1 year after the first BCG treatment and usually involves focal infection of the genitourinary tract (the site at which bacteria were introduced) and/or other sites that are typical for reactivation of mycobacterial disease, such as the vertebral spine or the retroperitoneal tissues . Noncaseating granulomas are found in the majority of cases, whether early or late . Most patients respond to treatment with antituberculous drugs; in early-presentation disease, when features of hypersensitivity predominate, glucocorticosteroids are sometimes added . Late localized infection often requires surgical resection.

Curr Microbiol, 2003 Feb, 46(2), 94 - 8
Cloning and characterization of a cytolytic and mosquito larvicidal delta-endotoxin from Bacillus thuringiensis subsp . darmstadiensis; Promdonkoy B et al.; The cytolytic delta-endotoxin gene from Bacillus thuringiensis subsp . darmstadiensis was amplified from genomic DNA by PCR by using primers designed from the sequence of cyt2Aa1 gene of B . thuringiensis subsp . kyushuensis . DNA sequence analysis revealed an open reading frame translating to a 259-amino acid sequence . The cloned gene was designated cyt2Aa2 . This gene was highly expressed in Escherichia coli as inclusion bodies that could be solubilized in 50 m M Na(2)CO(3), pH 10.5 . Activation of the solubilized protoxin by proteinase K (1% wt/wt, proteinase K/protoxin) yielded the active fragment of about 23 kDa . Cyt2Aa2 showed high hemolytic activity against sheep erythrocytes (hemolytic end- point 0.25 microgram/ml) and was toxic to Culex quinquefasciatus and Aedes aegypti larvae (LC(50) 0.5-1.0 microgram/ml).

Immunology, 2003 Jan, 108(1), 79 - 88
Mycobacterium bovis bacillus Calmette-Guérin-infected dendritic cells potently activate autologous T cells via a B7 and interleukin-12-dependent mechanism; Cheadle EJ et al.; Mycobacteria are potent adjuvants, can survive intracellularly and have been safely used for many years as vaccines against tuberculosis and leprosy . They are thus important potential vectors for recombinant vaccines . Many of their adjuvant properties are mediated following phagocytosis by dendritic cells (DC), which are in turn critical for priming naive T cells . Although the maturation of DC in response to mycobacteria, such as Mycobacterium bovis bacillus Calmette-Guerin (BCG), is well described the subsequent responses of autologous T cells to mycobacterium-infected DC remains uncharacterized . In our experiments DC infected with BCG expressed more co-stimulatory molecules than tumour-necrosis factor-alpha (TNF-alpha) -treated DC and stimulated more potent mixed leucocyte reactions . When autologous T cells were co-cultured with BCG-exposed DC they became highly activated, as determined by display of CD25, CD54 and CD71 on both CD4+ and CD8+ cells . In contrast, the response of T cells to TNF-alpha-matured DC was significantly less . Cytokine production from T cells cultured with BCG-exposed DC was enhanced with elevated secretion of interleukin-2 (IL-2), IL-10 and interferon-gamma (IFN-gamma) and was produced by both CD4+ and CD8+ lymphocytes as determined by intracellular staining . In particular, IFN-gamma secretion was increased from 50 pg/ml to 25 000 pg/ml and IL-10 secretion increased from 20 pg/ml to 300 pg/ml in BCG-exposed DC co-cultures . Blocking antibodies to B7.1 and B7.2 or IL-12 significantly reduced the secretion of IFN-gamma and reductions were also seen in the expression of CD25 and CD71 by CD4+ cells . These data demonstrate that mycobacterially infected DC are particularly potent activators of autologous T cells compared to TNF-alpha-exposed DC and that the resultant T cells are functionally superior.

Microbiol Immunol, 2002, 46(11), 773 - 5
Lethal factor to mice produced by Escherichia coli isolated from chickens with swollen head syndrome; Salvadori MR et al.; A lethal toxin similar to Bacillus cererus lethal toxin was detected in the culture supernatants of Escherichia coli isolated from chickens with swollen head syndrome . The lethal activity was heat-labile, protease-sensitive and killed mice within 10 min . The light microscopy of the histopathological studies revealed that the principal organ affected by this toxin was the lung but the liver and kidneys also showed lesions . The relevance of this lethal activity from E . coli remains to be determined.

J Exp Med, 2003 Jan 6, 197(1), 7 - 17
Mycobacteria target DC-SIGN to suppress dendritic cell function; Geijtenbeek TB et al.; Mycobacterium tuberculosis represents a world-wide health risk and immunosuppression is a particular problem in M . tuberculosis infections . Although macrophages are primarily infected, dendritic cells (DCs) are important in inducing cellular immune responses against M . tuberculosis . We hypothesized that DCs represent a target for M . tuberculosis and that the observed immuno-suppression results from modulation of DC functions . We demonstrate that the DC-specific C-type lectin DC-SIGN is an important receptor on DCs that captures and internalizes intact Mycobacterium bovis bacillus Calmette-Guerin (BCG) through the mycobacterial cell wall component ManLAM . Antibodies against DC-SIGN block M . bovis BCG infection of DCs . ManLAM is also secreted by M . tuberculosis-infected macrophages and has been implicated as a virulence factor . Strikingly, ManLAM binding to DC-SIGN prevents mycobacteria- or LPS-induced DC maturation . Both mycobacteria and LPS induce DC maturation through Toll-like receptor (TLR) signaling, suggesting that DC-SIGN, upon binding of ManLAM, interferes with TLR-mediated signals . Blocking antibodies against DC-SIGN reverse the ManLAM-mediated immunosuppressive effects . Our results suggest that M . tuberculosis targets DC-SIGN both to infect DCs and to down-regulate DC-mediated immune responses . Moreover, we demonstrate that DC-SIGN has a broader pathogen recognition profile than previously shown, suggesting that DC-SIGN may represent a molecular target for clinical intervention in infections other than HIV-1.

J Bacteriol, 2003 Jan, 185(2), 679 - 82
Identification of MpaA, an amidase in Escherichia coli that hydrolyzes the gamma-D-glutamyl-meso-diaminopimelate bond in murein peptides; Uehara T et al.; MpaA amidase was identified in Escherichia coli by its amino acid sequence homology with the ENP1 endopeptidase from Bacillus sphaericus . The enzymatic activity of MpaA, i.e., hydrolysis of the gamma-D-glutamyl-diaminopimelic acid bond in the murein tripeptide L-alanyl-gamma-D-glutamyl-meso-diaminopimelic acid, was demonstrated in the cell extract of a strain expressing mpaA from a multicopy plasmid . An mpaA mpl (murein peptide ligase) double mutant accumulated large amounts of murein tripeptide in its cytoplasm, consistent with the premise that MpaA degrades the tripeptide if its recycling via the peptidoglycan biosynthetic pathway is blocked.

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