Methods Find Exp Clin Pharmacol, 2002 Jul-Aug, 24(6), 371 - 91 Gateways to clinical trials; Bayes M et al.; Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses . The data in the following tables has been retrieved from the Clinical Studies knowledge area of Prous Science Integrity, the drug discovery and development portal, This issue focuses on the following selection of drugs: Aciclovir, alemtuzumab, alendronic acid sodium salt, alicaforsen sodium, alteplase, amifostine hydrate, antithymocyte globulin (equine), aspirin, atorvastatin calcium, azathioprine; Bacillus Calmette-Guerin, basiliximab, bicalutamide, bimatoprost, BMS-214662, brimonidine tartrate, buprenorphine hydrochloride; Cabergoline, carbamazepine, carboplatin, ciclosporine, cisplatin, cyclophosphamide; Daclizumab, desmopressin acetate, dihydroergotamine mesylate, dorzolamide hydrochloride, doxorubicin, dutasteride; Everolimus; Fluocinolone acetonide, frovatriptan, FTY-720, fulvestrant; Gabapentin, galantamine hydrobromide, ganciclovir, gemcitabine, glatiramer acetate; Hydrocodone bitartrate; Interferon beta, interferon beta-1a, interferon beta-1b, ipratropium bromide; Ketotifen; Lamivudine, latanoprost, levodopa, lidocaine hydrochloride, lonafarnib; Metformin hydrochloride, methylprednisolone, metoclopramide hydrochloride, mirtazapine, mitoxantrone hydrochloride, modafinil, muromonab-CD3, mycophenolate mofetil; NS-2330; Olopatadine hydrochloride, omalizumab, oxcarbazepine, oxycodone hydrochloride; Paclitaxel, paracetamol, piribedil, pramipexole hydrochloride, pravastatin sodium, prednisone; Quetiapine fumarate; Raloxifene hydrochloride, rituximab, rizatriptan sulfate, Ro-63-8695, ropinirole hydrochloride, rosiglitazone maleate; Simvastatin, siplizumab, sirolimus; Tacrolimus, tegaserod maleate, timolol maleate, tiotropium bromide, tipifarnib, tizanidine hydrochloride, tolterodine tartrate, topiramate, travoprost; Unoprostone isopropyl ester; Valganciclovir hydrochloride, visilizumab; Zidovudine. Plant Physiol, 1997 Apr, 113(4), 1071 - 1080 Extracellular Matrix Assembly in Diatoms (Bacillariophyceae) (II . 2,6-Dichlorobenzonitrile Inhibition of Motility and Stalk Production in the Marine Diatom Achnanthes longipes); Wang Y et al.; The cellulose synthesis inhibitor 2,6-dichlorobenzonitrile (DCB) and the DCB analogs 2-chloro-6-fluorobenzonitrile, 3-amino-2,6-dichlorobenzonitrile, and 5-dimethylamino-naphthalene-1-sulfonyl-(3-cyano-2, 4-dichloro)aniline (DCBF) inhibited extracellular adhesive production in the marine diatom Achnanthes longipes, resulting in a loss of motility and a lack of permanent adhesion . The effect was fully reversible upon removal of the inhibitor, and cell growth was not affected at concentrations of inhibitors adequate to effectively interrupt the adhesion sequence . Video microscopy revealed that the adhesion sequence was mediated by the export and assembly of polymers, and consisted of initial attachment followed by cell motility and eventual production of permanent adhesive structures in the form of stalks that elevated the diatom above the substratum . A . longipes adhesive polymers are primarily composed of noncellulosic polysaccharides (B.A . Wustman, M.R . Gretz, and K.D . Hoagland {1997} Plant Physiol 113: 1059-1069) . These results, together with the discovery of DCB inhibition of extracellular matrix assembly in noncellulosic red algal unicells (S.M . Arad, O . Dubinsky, and B . Simon {1994} Phycologia 33: 158-162), indicate that DCB inhibits synthesis of noncellulosic extracellular polysaccharides . A fluorescent probe, DCBF, was synthesized and shown to inhibit adhesive polymer production in the same manner as DCB . DCBF specifically labeled an 18-kD polypeptide isolated from a membrane fraction . Inhibition of adhesion by DCB and its analogs provides evidence of a direct relationship between polysaccharide synthesis and motility and permanent adhesion. Plant Physiol, 1997 Apr, 113(4), 1059 - 1069 Extracellular Matrix Assembly in Diatoms (Bacillariophyceae) (I . A Model of Adhesives Based on Chemical Characterization and Localization of Polysaccharides from the Marine Diatom Achnanthes longipes and Other Diatoms); Wustman BA et al.; Extracellular adhesives from the diatoms Achnanthes longipes, Amphora coffeaeformis, Cymbella cistula, and Cymbella mexicana were characterized by monosaccharide and methylation analysis, lectin-fluorescein isothiocyanate localization, and cytochemical staining . Polysaccharide was the major component of adhesives formed during cell motility, synthesis of a basal pad, and/or production of a highly organized shaft . Hot water-insoluble/hot 0.5 M NaHCO3-soluble anionic polysaccharides from A . longipes and A . coffeaeformis adhesives were primarily composed of galactosyl (64-70%) and fucosyl (32-42%) residues . In A . longipes polymers, 2,3-, t-, 3-, and 4-linked/substituted galactosyl, t-, 3-, 4-, and 2-linked fucosyl, and t- and 2-linked glucuronic acid residues predominated . Adhesive polysaccharides from C . cistula were EDTA-soluble, sulfated, consisted of 83% galactosyl (4-, 4,6-, and 3,4-linked/substituted) and 13% xylosyl (t-, 4f/5p-, and 3p-linked/substituted) residues, and contained no uronosyl residues . Ulex europaeus agglutinin uniformly localized {alpha}(1,2)-L-fucose units in C . cistula and Achnanthes adhesives formed during motility and in the pads of A . longipes . D-Galactose residues were localized throughout the shafts of C . cistula and capsules of A . coffeaeformis . D-Mannose and/or D-glucose, D-galactose, and {alpha}(t)-L-fucose residues were uniformly localized in the outer layers of A . longipes shafts by Cancavalia ensiformis, Abrus precatorius, and Lotus tetragonolobus agglutinin, respectively . A model for diatom cell adhesive structure was developed from chemical characterization, localization, and microscopic observation of extracellular adhesive components formed during the diatom cell-attachment process. Proc Natl Acad Sci U S A, 2002 Sep 17, 99(19), 12120 - 5 Epub 2002 Sep 09. Crystal structure of DhbE, an archetype for aryl acid activating domains of modular nonribosomal peptide synthetases; May JJ et al.; The synthesis of the catecholic siderophore bacillibactin is accomplished by the nonribosomal peptide synthetase (NRPS) encoded by the dhb operon . DhbE is responsible for the initial step in bacillibactin synthesis, the activation of the aryl acid 2,3-dihydroxybenzoate (DHB) . The stand-alone adenylation (A) domain DhbE, the structure of which is presented here, exhibits greatest homology to other NRPS A-domains, acyl-CoA ligases and luciferases . It's structure is solved in three different states, without the ligands ATP and DHB (native state), with the product DHB-AMP (adenylate state) and with the hydrolyzed product AMP and DHB (hydrolyzed state) . The 59.9-kDa protein folds into two domains, with the active site at the interface between them . In contrast to previous proposals of a major reorientation of the large and small domains on substrate binding, we observe only local structural rearrangements . The structure of the phosphate binding loop could be determined, a motif common to many adenylate-forming enzymes, as well as with bound DHB-adenylate and the hydrolyzed product DHB*AMP . Based on the structure and amino acid sequence alignments, an adapted specificity conferring code for aryl acid activating domains is proposed, allowing assignment of substrate specificity to gene products of previously unknown function. J Mol Biol, 2002 Aug 23, 321(4), 693 - 701 The unfolding of oxidized c-type cytochromes: the instructive case of Bacillus pasteurii; Bartalesi I et al.; The reversible unfolding of oxidized Bacillus pasteurii cytochrome c(553) by guanidinium chloride under equilibrium conditions has been monitored by NMR and optical spectroscopy . The results obtained indicate that unfolding takes place through a mechanism involving the detachment from heme iron coordination of the sulfur of the Met71 axial ligand and yielding either a high spin (HS) or a low spin (LS(1)) species, depending on the pH value . In the LS(1) form the Met71 is replaced by another protein ligand, possibly Lys . The ligand exchange reaction does not reach completion until the protein backbone reaches a largely unfolded state, as monitored through 1H-15N NMR experiments, thus demonstrating that there is a significant correlation between formation of the Fe-S bond and native structure stability . 1H/2H exchange data, however, show that helix alpha(3), the C-terminal region of helix alpha(4), and helix alpha(5) maintain low exchangeability of the amide protons in the LS(1) form . This finding most likely implies that these regions maintain some ordered non-covalent structure, in which the amide moieties are involved in H-bonds . Finally, a folding mechanism is proposed and discussed in terms of analogies and differences with the larger mitochondrial cytochrome c proteins . It is concluded that the thermodynamic stability of the region around the metal cofactor is determined by the chemical nature of the residues around the axial methionine residue. Prog Nucleic Acid Res Mol Biol, 2002, 72, 1 - 39 Viral strategies of translation initiation: ribosomal shunt and reinitiation; Ryabova LA et al.; Due to the compactness of their genomes, viruses are well suited to the study of basic expression mechanisms, including details of transcription, RNA processing, transport, and translation . In fact, most basic principles of these processes were first described in viral systems . Furthermore, viruses seem not to respect basic rules, and cases of "abnormal" expression strategies are quiet common, although such strategies are usually also finally observed in rare cases of cellular gene expression . Concerning translation, viruses most often violate Kozak's original rule that eukaryotic translation starts from a capped monocistronic mRNA and involves linear scanning to find the first suitable start codon . Thus, many viral cases have been described where translation is initiated from noncapped RNA, using an internal ribosome entry site . This review centers on other viral translation strategies, namely shunting and virus-controlled reinitiation as first described in plant pararetroviruses (Caulimoviridae) . In shunting, major parts of a complex leader are bypassed and not melted by scanning ribosomes . In the Caulimoviridae, this process is coupled to reinitiation after translation of a small open reading frame; in other cases, it is possibly initiated upon pausing of the scanning ribosome . Most of the Caulimoviridae produce polycistronic mRNAs . Two basic mechanisms are used for their translation . Alternative translation of the downstream open reading frames in the bacilliform Caulimoviridae occurs by a leaky scanning mechanism, and reinitiation of polycistronic translation in many of the icosahedral Caulimoviridae is enabled by the action of a viral transactivator . Both of these processes are discussed here in detail and compared to related processes in other viruses and cells. Nat Med, 2002 Oct, 8(10), 1171 - 4 Epub 2002 Sep 09. A pantothenate auxotroph of Mycobacterium tuberculosis is highly attenuated and protects mice against tuberculosis; Sambandamurthy VK et al.; With the advent of HIV and the widespread emergence of drug-resistant strains of Mycobacterium tuberculosis, newer control strategies in the form of a better vaccine could decrease the global incidence of tuberculosis . A desirable trait in an effective live attenuated vaccine strain is an ability to persist within the host in a limited fashion in order to produce important protective antigens in vivo . Attenuated M . tuberculosis vaccine candidates have been constructed by deleting genes required for growth in mice . These candidate vaccines did not elicit adequate protective immunity in animal models, due to their inability to persist sufficiently long within the host tissues . Here we report that an auxotrophic mutant of M . tuberculosis defective in the de novo biosynthesis of pantothenic acid (vitamin B5) is highly attenuated in immunocompromised SCID mice and in immunocompetent BALB/c mice . SCID mice infected with the pantothenate auxotroph survived significantly longer (250 days) than mice infected with either bacille Calmette-Guerin (BCG) vaccine or virulent M . tuberculosis (77 and 35 days, respectively) . Subcutaneous immunization with this auxotroph conferred protection in C57BL/6J mice against an aerosol challenge with virulent M . tuberculosis, which was comparable with that afforded by BCG vaccination . Our findings highlight the importance of de novo pantothenate biosynthesis in limiting the intracellular survival and pathogenesis of M . tuberculosis without reducing its immunogenicity in vaccinated mice. Int Arch Allergy Immunol, 2002 Aug, 128(4), 280 - 91 Bioinformatic methods for allergenicity assessment using a comprehensive allergen database; Hileman RE et al.; BACKGROUND: A principal aim of the safety assessment of genetically modified crops is to prevent the introduction of known or clinically cross-reactive allergens . Current bioinformatic tools and a database of allergens and gliadins were tested for the ability to identify potential allergens by analyzing 6 Bacillus thuringiensis insecticidal proteins, 3 common non-allergenic food proteins and 50 randomly selected corn (Zea mays) proteins . METHODS: Protein sequences were compared to allergens using the FASTA algorithm and by searching for matches of 6, 7 or 8 contiguous identical amino acids . RESULTS: No significant sequence similarities or matches of 8 contiguous amino acids were found with the B . thuringiensis or food proteins . Surprisingly, 41 of 50 corn proteins matched at least one allergen with 6 contiguous identical amino acids . Only 7 of 50 corn proteins matched an allergen with 8 contiguous identical amino acids . When assessed for overall structural similarity to allergens, these 7 plus 2 additional corn proteins shared >or=35% identity in an overlap of >or=80 amino acids, but only 6 of the 7 were similar across the length of the protein, or shared >50% identity to an allergen . CONCLUSIONS: An evaluation of a protein by the FASTA algorithm is the most predictive of a clinically relevant cross-reactive allergen . An additional search for matches of 8 amino acids may provide an added margin of safety when assessing the potential allergenicity of a protein, but a search with a 6-amino-acid window produces many random, irrelevant matches . J Bacteriol, 2002 Oct, 184(19), 5410 - 7 Loss of catabolite repression function of HPr, the phosphocarrier protein of the bacterial phosphotransferase system, affects expression of the cry4A toxin gene in Bacillus thuringiensis subsp . israelensis; Khan SR et al.; HPr, the phosphocarrier protein of the bacterial phosphotransferase system, mediates catabolite repression of a number of operons in gram-positive bacteria . In order to participate in the regulatory process, HPr is activated by phosphorylation of a conserved serine-46 residue . To study the potential role of HPr in the regulation of Cry4A protoxin synthesis in Bacillus thuringiensis subsp . israelensis, we produced a catabolite repression-negative mutant by replacing the wild-type copy of the ptsH gene with a mutated copy in which the conserved serine residue of HPr was replaced with an alanine . HPr isolated from the mutant strain was not phosphorylated at Ser-45 by HPr kinase, but phosphorylation at His-14 was found to occur normally . The enzyme I and HPr kinase activities of the mutant were not affected . Analysis of the B . thuringiensis subsp . israelensis mutant harboring ptsH-S45A in the chromosome showed that cry4A expression was derepressed from the inhibitory effect of glucose . The mutant strain produced both cry4A and sigma(35) gene transcripts 4 h ahead of the parent strain, but there was no effect on sigma(28) synthesis . In wild-type B . thuringiensis subsp . israelensis cells, cry4A mRNA was observed from 12 h onwards, while in the mutant it appeared at 8 h and was produced for a longer period . The total amount of cry4A transcripts produced by the mutant was higher than by the parent strain . There was a 60 to 70% reduction in the sporulation efficiency of the mutant B . thuringiensis subsp . israelensis strain compared to the wild-type strain. J Econ Entomol, 2002 Aug, 95(4), 832 - 7 Evaluation of the natural refuge function for Helicoverpa arnigera (Lepidoptera: Noctuidae) within Bacillus thuringiensis transgenic cotton growing areas in north China; Wu K et al.; The density of Helicoverpa armigera (Hubner) populations on Bacillus thuringiensis Berliner (Bt) transgenic cotton, corn, peanut, and soybean; differences in its development on Bt cotton and common (nontransgenic) cotton; and the potential for mating among populations from Bt cotton fields and other crop fields were investigated in the suburbs of Xinxiang City (Henan Province) and Langfang City (Hebei Province) in the southern and northern parts of north China, respectively . Although development of H . armigera on Bt cotton was much slower than on common cotton, there was a still high probability of mating between populations from Bt cotton and other sources due to the scattered emergence pattern of H . armigera adults, and overlap of the second and third generations . In a cotton and corn growing region, early and late planted corn provided suitable refugia for the third and fourth generations of H . armigera, but not for the second generation . In a cotton and soybean/ peanut mix system, noncotton crops provided a natural refugia from the second- to fourth-generation H . armigera, but function of the refuge would closely depend on the proportion of Bt cotton . Consequently, it may be necessary to compensate the original mixed cropping patterns in different areas for delaying resistance development of H . armigera to Bt cotton. J Econ Entomol, 2002 Aug, 95(4), 826 - 31 Resistance monitoring of Helicoverpa armigera (Lepidoptera Noctuidae) to Bacillus thuringiensis insecticidal protein in China; Wu K et al.; Sensitivities of Helicoverpa armigera (Hubner) field populations to Bacillus thuringiensis (Bt) insecticidal protein CrylAc were monitored during 1998-2000 in China . A total of 41 strains was sampled, and most of them were collected from Bt cotton planting regions . The range of IC50 values (concentration producing 50% inhibition of larval development to third instar) among different populations in 1998,1999, and 2000 were 0.020-0.105 microg/ml, 0.016-0.099 microg/ml, and 0.016-0.080 microg/ml, respectively . Diagnostic concentration studies (IC99) showed that the percentage of individuals reaching third instar ranged from 0 to 4.35%, with only eight of the 41 tested populations showing values above 0% . Also interesting was a trend over successive years in which fewer populations contained individuals that survived the diagnostic concentration by reaching third instar . Considering these data, it was determined that the field populations sampled during the 3-yr study were susceptible to CrylAc protein, and that movement toward resistance among H . armigera populations was not apparent. J Econ Entomol, 2002 Aug, 95(4), 763 - 9 Behavior of bollworm (Lepidoptera: Noctuidae) larvae on genetically engineered cotton; Gore J et al.; Reports of bollworm, Helicoverpa zea (Boddie), larvae feeding in white flowers of Bollgard cotton have been relatively common since the commercialization of this technology in 1996 . Field studies were conducted in Louisiana to determine if differences in bollworm larval behavior occuron non-Bollgard (cultivar 'Deltapine 5415') and Bacillus thuringiensis (Bt), Bollgard ('NuCOTN 33B') cottons . Larvae were placed on the terminal foliage of either single cotton plants or on all plants within 1-m row micro-plots . On preflowering cotton plants, significantly more bollworms moved from the site of infestation (terminal) on Bollgard plants compared with that on non-Bollgard plants . On individual flowering plants, the number of nodes larvae moved from the terminal and the number of infested bolls was greater on Bollgard cotton plants . Similar differences between Bollgard and non-Bollgard plants in the percentage of infested terminals and squares were observed at 48-h after infestation when 1-m rows were infested . These data will be used to refine scouting protocols for bollworm larvae on Bollgard cotton. Acta Cient Venez, 2002, 53(1), 7 - 14 {Diatoms from Unare and Píritu Lagoons, Anzoátegui state, Venezuela . Subclass Bacillariophycidae D . G . Mann }; Subero-Pino SS et al.; The diatom flora of Unare and Piritu Lagoons, Venezuela, comprises freshwater, estuarine and marine organisms . In this paper, 15 species of diatoms belonging to the subclass Bacillariophyceae D.G . Mann are described . Water samples from Unare and Piritu lagoons (Venezuela) were collected from November 1988 to July 1989 . Samples were fixed with neutral formaldehyde (10%) and were examined under phase contrast microscopy . Species under study were Dictyoneis marginata (Lewis) Cleve, Cymbella affinis (Kutzing) Cleve, Cymbella sp., Achnanthes brevipes var . intermedia (Kutzing) Cleve, Lyrella irroratoides (Hustedt) Mann, Navicula carinifera (Grunow) Peragallo, Navicula liber (W . Smith) Peragallo, Navicula vacillans (Schmidt) Peragallo, Navicula sp, Amphora crassa var . campechiana Grunow, Amphora decussata Grunow, Caloneis powelli (Lewis) Cholnoky, Tryblionella acuta (Cleve) Mann in Round et al., Tryblionella apiculata Gregory, Surirella febigerii Lewis. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1997, 29(4), 356 - 362 Purification and Characterization of DNA Helicase BstH2 from Bacillus Stearothermophilus; Zhang XH et al.; In the purification of DNA helicase BstH1 we have partially purified the second DNA helicase BstH2 from Bacillus Stearothermophilus through Polymin P precipitation, ammonia sulfate precipitation and chromatographic steps with DEAE-cellulose, phosphocellulose, Blue-Sepharose, FPLC Superose 12, Mono Q and second Mono Q . The ATPase activity of BstH2 depends on Mg(2+) and is differentially stimulated by different types of nucleic acids . BstH2 has a maximal ATPase activity at 55 degrees . The ATPase activity is greatly inhibited by E . coli SSB or higher ionic strength . The DNA helicase activity of BstH2 depends on ATP and Mg(2+) . BstH2 can unwind partial duplex DNA as well as blunt-ended duplex DNA . E . coli SSB stimulates the unwinding reaction catalyzed by BstH2. J Egypt Soc Parasitol, 2002 Aug, 32(2), 415 - 24 Comparative study on the effect of two Bacillus thuringiensis strains of the same serotype on three coleopteran pests of stored wheat; Abdel-Razek AS; The biological control of insect populations of stored products by insect pathogens such as, Bacillus thuringiensis (Berliner) is considered as an alternative to synthetic insecticides . So far, only B . thuringiensis among microbial pesticides have been approved for application against stored product insect pests . In this study we used a laboratory culture from an indigenous isolate of B . thuringiensis subsp . morrisoni, and the commercial formulation, B . thuringiensis subsp . tenebrionis . The bioassays with adults of the three beetle species (Rhyzopertha dominica (F.), Silophilus granarius (L.), and Tribolium confusum (Jacqueline du Val) and larvae of T . confusum were done at four different concentrations 3000, 1000, 500, and 250 ppm . Mortality records were made after 7, 14, 21, 28 days of exposure . Among the strains tested, the indigenous isolate B . thuringiensis subsp . morrisoni gave the highest mortality for adults of R . dominica and T . confusum after 21 and 28 days of exposure . It was (67, 78%) and (43, 65%), respectively . Larvae of T . confusum showed that the isolate, morrisoni was the most active where it gave mortality records of 76, 60, 55 and 35% at 3000, 1000, 500 and 250 ppm after 28 days of exposure, while the other strain did not show any actual mortality . This demonstrates the possibility of using B . thuringiensis subsp . Morrisoni for the protection of stored grains from pest infestation. Mol Immunol, 2002 Sep, 39(1-2), 113 - 9 Development of new vaccines and diagnostic reagents against tuberculosis; Mustafa AS; Tuberculosis (TB) is a major infectious disease problem with one-third of the world population infected, 8 million people developing the active disease and 2 million dying of TB each year . The attenuated Mycobacterium bovis Bacillus Calmette Guerin (BCG) is the only available vaccine against TB . However, the trials conducted in different parts of the world have shown that this vaccine doe not provide consistent protection against TB . The purified protein derivative (PPD) of Mycobacterium tuberculosis is the commonly used reagent for the diagnosis of TB . However, PPD lacks specificity because of the presence of antigens crossreactive with M . bovis BCG and other mycobacteria . The studies to identify M . tuberculosis antigens and epitopes as candidates for new protective vaccines and specific diagnostic reagents against TB have led to the identification and characterization of several major antigens of M . tuberculosis including heat shock proteins (hsp) and secreted antigens present in the culture filtrate (CF) of M . tuberculosis . Some of these antigens have shown promise as new candidate vaccines (hsp60, Ag85 and ESAT-6, etc.) and specific diagnostic reagents (ESAT-6 and CFP10, etc.) for TB . Moreover, in the mouse model of TB, vaccination with DNA-hsp60 has immunotheraputic effects and helps in eradication of persisters . In addition, identification of proper adjuvant and delivery systems has shown the promise to overcome the problem of poor immunogenicity associated with subunit and peptide based vaccines . More recently, the comparison of the genome sequence of M . tuberculosis with M . bovis BCG and other mycobacteria has led to the identification of M . tuberculosis-specific genomic regions . Evaluation of these regions for encoding proteins with immunological reactivity can lead to the identification of additional antigens of M . tuberculosis useful as new vaccines and reagents for specific diagnosis of TB. Insect Biochem Mol Biol, 2002 Sep, 32(9), 1025 - 36 Cry1A toxins of Bacillus thuringiensis bind specifically to a region adjacent to the membrane-proximal extracellular domain of BT-R(1) in Manduca sexta: involvement of a cadherin in the entomopathogenicity of Bacillus thuringiensis; Dorsch JA et al.; Many subspecies of the soil bacterium Bacillus thuringiensis produce various parasporal crystal proteins, also known as Cry toxins, that exhibit insecticidal activity upon binding to specific receptors in the midgut of susceptible insects . One such receptor, BT-R(1) (210 kDa), is a cadherin located in the midgut epithelium of the tobacco hornworm, Manduca sexta . It has a high binding affinity (K(d) approximately 1nM) for the Cry1A toxins of B . thuringiensis . Truncation analysis of BT-R(1) revealed that the only fragment capable of binding the Cry1A toxins of B . thuringiensis was a contiguous 169-amino acid sequence adjacent to the membrane-proximal extracellular domain . The purified toxin-binding fragment acted as an antagonist to Cry1Ab toxin by blocking the binding of toxin to the tobacco hornworm midgut and inhibiting insecticidal action . Exogenous Cry1Ab toxin bound to intact COS-7 cells expressing BT-R(1) cDNA, subsequently killing the cells . Recruitment of BT-R(1) by B . thuringiensis indicates that the bacterium interacts with a specific cell adhesion molecule during its pathogenesis . Apparently, Cry toxins, like other bacterial toxins, attack epithelial barriers by targeting cell adhesion molecules within susceptible insect hosts. Intern Med, 2002 Aug, 41(8), 671 - 3 Successful non-surgical treatment of brain abscess and necrotizing fasciitis caused by Bacillus cereus; Mori T et al.; Musculoskeletal and central nervous system infections caused by Bacillus cereus are very rare . Only a few cases have been reported, whose clinical courses strongly suggested that surgical procedures combined with appropriate antimicrobial therapy are necessary to cure these infections . A 60-year-old man with severe neutropenia due to myelodysplastic syndrome, developing necrotizing fasciitis and brain abscess caused by Bacillus cereus is reported . Without performing any surgical procedures, the patient was successfully treated with systemic antimicrobial therapy combined with granulocyte colony stimulating factor, which contributed to the increase in the neutrophil count. Comp Med, 2002 Aug, 52(4), 322 - 7 Characterization of lymphocyte subsets in the bronchiolar lymph nodes of BALB/c mice infected with cilia-associated respiratory bacillus; Kendall LV et al.; Cilia-associated respiratory (CAR) bacillus is an unclassified, gram-negative, extracellular bacterium that causes chronic respiratory tract disease in rodents . Infected mice develop microscopic lesions characterized by a primary lymphocytic response followed by macrophage and neutrophilic infiltration . To characterize the lymphocytic subsets that respond to CAR bacillus infection, BALB/c mice were inoculated with 10(5) CAR bacillus bacteria . At seven weeks after inoculation, mice were euthanized and the tracheobronchiolar and hilar lymph nodes were collected and stained for cell surface markers to T cells (CD3, CD4, and CD8), B cells (B220, CD5), natural killer (NK) cells (pan-NK) and intracellular interleukin 10 (IL-10) and interferon-gamma (IFN-gamma) . Flow cytometric analysis of lymph nodes from CAR bacillus-infected mice revealed 11% increase in frequency of B cells (R220+), 12% increase in the frequency of double-negative (CD4-CD8-CD3+) T cells, and slight increase in the B-1 subset of B cells (B220+CD5+) . There was no change in the frequency of NK cells . The CAR bacillus-infected mice had an overall decrease in the frequency of T cells . Intracellular cytokine staining revealed distinct populations of T cells producing IL-10 and IFN-gamma, and IL-10 production from B cells; NK cells were not a substantial source of IFN-gamma . To our knowledge, this is the first characterization of lymphocytic responses and suggestion that B cells and double-negative T cells may be principally responsible for the lesions associated with CAR bacillus infection. Mol Reprod Dev, 2002 Sep, 63(1), 89 - 95 Centrosome dynamics and inheritance in related sexual and parthenogenetic Bacillus (Insecta Phasmatodea); Marescalchi O et al.; In animals, some general features of centrosome dynamics and inheritance have been widely recognized . The most acknowledged model assigns to sperm the contribution of a centriole to the fertilized egg, which in turn provides the pericentriolar materials, including gamma-tubulin, recruiting them from the cytoplasm: the main zygote microtubule organizing center (MTOC) is thus reconstituted to organize first the spermaster and then the full first embryonic spindle . Obviously the model cannot apply to parthenogenetic systems, which actually rely on egg components alone . In stick insects of the Bacillus genus, the spindle of both somatic and germ cells is clearly anastral, therefore we have been investigating their centrosome in sexual and parthenogenetic taxa by analyzing its component dynamics and transmission through the use of monoclonal beta- and gamma-tubulin antibodies and transmission electron microscopy (TEM) . It has been shown that in sexually reproducing species the spermatozoon does not contribute the centriole, so that the egg wholly provides the MTOC and the ensuing anastral spindle of the embryo: MTs appear to derive from pronuclear chromatin surroundings and no asters are observed . The parthenogenetic embryo development is the same as the sexual one if syngamy is excepted . The parthenogenetic mechanism realized by these panoistic insects appears to differ from that observed in the meroistic hymenopteran and drosophilid species, where the embryo spindle derives from asters formed in the egg cortex . In stick insects, the lack of sperm contribution to embryonic centrosome appears to be a major trait accounting for the widespread occurrence of facultative and obligate parthenogenesis within the order . Chembiochem, 2002 Sep 2, 3(9), 866 - 73 Specificity studies of bacillus 1,3-1,4-beta-glucanases and application to glycosynthase-catalyzed transglycosylation; Fairweather JK et al.; Bacillus 1,3-1,4-beta-glucanases hydrolyze 1,3-1,4-beta-gluco-oligosaccharides with a retaining mechanism . The binding-site cleft of these endoglycosidases is composed of six subsites (-4 to +2) of which subsite -3 makes the largest contribution to transition state stabilization . The specificity of this subsite is here analyzed for both glycosidase and glycosynthase activities in the wild-type and the nucleophile-less E134A mutant Bacillus licheniformis enzymes . A D-galactosyl residue on the nonreducing end of a trisaccharide substrate is accepted by the enzyme and binds at subsite -3 in the productive enzyme-substrate complex . The wild-type enzyme catalyzes the hydrolysis of the substrate Glcbeta4Glcbeta3GlcbetaMU (Glc=glucosyl, MU=4-methylumbelliferyl) with a k(cat)/K(M) value only 1.3-fold higher than for the Galbeta4Glcbeta3GlcbetaMU (Gal=galactosyl) substrate . The corresponding alpha-fluorides act as good donors for the glycosynthase condensation reaction with mono- and disaccharide acceptors catalyzed by the E134A mutant . Whereas self-condensation and elongation products are also obtained as minor compounds with the Glcbeta4Glcbeta3GlcalphaF donor, nearly quantitative yields of single condensation products are obtained with the Galbeta4Glcbeta3GlcalphaF donor, in which the axial configuration of the 4-OH group on the nonreducing end prevents self-condensation and elongation reactions. J Agric Food Chem, 2002 Sep 11, 50(19), 5429 - 36 Physicochemical and nitrogen solubility properties of Bacillus proteinase hydrolysates of sodium caseinate incubated with transglutaminase pre- and post-hydrolysis; Flanagan J et al.; Sodium caseinate (NaCN), hydrolyzed with Protamex, a Bacillus proteinase preparation, to 0.5, 1.3, and 17.5% degrees of hydrolysis, was incubated with transglutaminase (TGase) for 3, 42, and 290 min at enzyme/substrate ratios of 1, 1, and 10% (w/w), respectively, pre- and post-hydrolysis . The electrophoretic, reversed-phase high-performance liquid chromatography (RP-HPLC) and nitrogen solubility profiles of the modified products were investigated . Combinations of hydrolysis and incubation with TGase generated products displaying novel physicochemical and nitrogen solubility properties . Significant changes in sodium dodecyl sulfate (SDS) and urea polyacrylamide gel electrophoresis profiles were apparent in the modified caseinate samples . Extensive TGase cross-linking resulted in polymers that were unable to enter the resolving gel during SDS polyacrylamide gradient gel electrophoresis . Extensive combined enzymatic modification resulted in peptides eluting earlier on RP-HPLC than limited combined enzymatic modification or limited hydrolysis . Combination of enzymatic treatments resulted in significantly (P < 0.005) improved solubility around pH 4.6, compared to incubation with TGase or hydrolysis of NaCN alone. Clin Diagn Lab Immunol, 2002 Sep, 9(5), 1067 - 71 Immunoglobulin G antibody response to infection with coccoid forms of Helicobacter pylori; Figueroa G et al.; An increasing number of studies support a potential role for coccoid forms in Helicobacter pylori infection . Evidence for this was obtained through scanning microscopy, genetic analysis for virulence traits, examination of the presence and activity of key enzymes, and other methods . We studied the serum immunoglobulin G responses to coccoid H . pylori forms by enzyme-linked immunosorbent assay (ELISA) and immunoblotting and compared them with those of bacillary cells . Sera from a total of 295 infected individuals were studied; these included sera from 100 patients with duodenal ulcers, 98 patients with nonulcer dyspepsia, 11 patients with gastroduodenal cancer, and 86 asymptomatic individuals . Initially, we characterized and selected coccoid and bacillary antigenic preparations by one-dimensional (1-D) and 2-D gel electrophoresis and immunoblotting . Data showed that coccoid and bacillary preparations with comparable protein contents have similar patterns in 1-D and 2-D electrophoresis gels and antigenic recognition at blotting . These results revealed that coccoid and spiral antigens in ELISA can equally recognize specific antibodies to H . pylori in sera from infected individuals . The analysis of the spiral and coccoid preparations by Western blotting showed no major differences in antigen recognition . No specific bands or profiles associated with a single gastric condition were identified. Clin Diagn Lab Immunol, 2002 Sep, 9(5), 1057 - 60 Bacillus species are present in chewing tobacco sold in the United States and evoke plasma exudation from the oral mucosa; Rubinstein I et al.; Five Bacillus species, predominantly Bacillus megaterium and Bacillus pumilus, were isolated from two popular brands of commercially available chewing tobacco {(5.0 +/- 1) x 10(6) CFU/ml of supernatant; results for four experiments} . Moreover, the supernatant of the Bacillus culture evoked plasma exudation from postcapillary venules in the intact hamster cheek pouch, exudation that was mediated by the kallikrein/kinin metabolic pathway . Taken together, these data indicate that Bacillus species contaminate chewing tobacco commercially available in the United States and elaborate a potent exogenous virulence factor(s) that injures the oral mucosa. Res Vet Sci, 2002 Oct, 73(2), 145 - 52 The efficacy of bacille Calmette-Guérin vaccine in wild brushtail possums (Trichosurus vulpecula); Corner LA et al.; A population of wild brushtail possums (Trichosurus vulpecula) in which bovine tuberculosis was endemic was vaccinated with live bacille Calmette-Guerin (BCG) to determine the efficacy of vaccination . The population on the 56 hectare site was monitored bimonthly over 2 years using a capture-release regime . During the study tuberculosis was diagnosed by clinical and post mortem examination . Possums were vaccinated with BCG by both intranasal aerosol and conjunctival instillation . Possums were revaccinated on average every 5 months . Over the 2 years, 300 possums were recruited to the study with 149 being allocated to the vaccination group . There were significantly fewer cases of tuberculosis in the vaccinated (4 cases) than in the unvaccinated group (13 cases; P=0.023) . The vaccine efficacy was 69% . An attempt was made to increase the incidence of disease by releasing onto the site possums that had been experimentally infected with a strain of M . bovis unknown in the area . However, this did not result in any additional cases . BCG vaccine was shown to have a level of efficacy which could be of assistance in controlling tuberculosis in wild possum populations . The future use of vaccination for the control of tuberculosis in wild possum populations is discussed. Acta Trop, 2002 Sep, 83(3), 241 - 7 Efficacy of a floating sustained release formulation of Bacillus thuringiensis ssp . israelensis in controlling Culex quinquefasciatus larvae in polluted water habitats; Gunasekaran K et al.; Larvicidal efficacy and residual activity of a floating sustained release formulation of Bacillus thuringiensis ssp . israelensis (Bti) was tested in polluted water habitats against Culex quinquefasciatus, the vector of bancroftian filariasis, breeding in cesspools and cesspits . A total of 25 habitats having 65 m(2) water surface area with C . quinquefasciatus immatures were treated with the formulation at the rate of 15 kg/ha . For comparison, in another area, 18 habitats with a water surface area of 70 m(2) were kept untreated . Evaluation was continued up to 179 days with three rounds of Bti application . The mean number of egg rafts did not differ significantly between treated and untreated habitats during the trial period indicating same level of recruitment . However, abundance of larvae and pupae was significantly lower in Bti treated habitats compared to that in untreated ones . The activity of the formulation, with 2 weeks delay in onset compared to treatment, caused >80% reduction in pupal abundance for a period of 30-34 days . Considering this, it is suggested that monthly application of Bti formulation will considerably reduce the recruitment of pupae of C . quinquefasciatus. J Biol Inorg Chem, 2002 Sep, 7(7-8), 704 - 12 Epub 2002 Mar 22. Structural determinants and hydrogen-bond network of the mononuclear zinc(II)-beta-lactamase active site; Dal Peraro M et al.; Zinc(II)-beta-lactamases are among the latest generation of antibiotic-resistant enzymes developed by bacteria against beta-lactams . Here we have used density functional theory to provide the full structure of the catalytic site from Bacillus cereus mononuclear beta-lactamase II . Calculations are carried out on relative large models built on the X-ray structure of the free enzyme at the highest available resolution (1.7 A, PDB entry 3BC2) . The most stable conformation emerging from our calculations consists of a Zn(II)-bound hydroxide, which acts as nucleophilic agent in the enzymatic reaction, highly stabilized by a complex hydrogen-bond network, in which the protonation state of Asp90 plays a major role . The pattern differs from that previously proposed on the basis of smaller models . Furthermore, the calculations confirm that Arg91 contributes to determine the orientation and the protonation state of Asp90, as recently suggested by mutagenesis experiments. Urol Res, 2002 Sep, 30(4), 233 - 9 Epub 2002 Jul 05. The role of LFA-1 in the lysis of bladder cancer cells by bacillus Calmette-Guérin and interleukin 2-activated killer cells; Suttmann H et al.; Activated cytotoxic effector cells such as bacillus Calmette-Guerin (BCG)-activated killer (BAK) and lymphokine-activated killer (LAK) cells are thought to mediate the antitumor effects in the immunotherapy of superficial bladder cancer with BCG . We investigated the role of the leukocyte-function-antigen-1 and its two subunits CD11a and CD18 in the lysis of bladder tumor cells by both effector cell populations . We used flow cytometry to measure CD11a and CD18 expression on BAK and LAK cells . The involvement of both surface molecules in the lysis of bladder tumor cells was determined by phase contrast microscopy and a set of radioactive-release assays using specific inhibitory antibodies . BAK and LAK cells expressed more CD11a and CD18 on their surfaces than unstimulated peripheral blood mononuclear cells . Effector-target cell adhesion was a prerequisite for the cell-mediated cytotoxicity of BAK and LAK cells against bladder tumor targets . Cytotoxicity of both BAK and LAK cells was inhibited by a combination of anti-CD11a and -CD18 monoclonal antibodies . Our study gives further insight into the complex interactions of the adhesion molecules of activated immune cells and their respective tumor targets and might help to increase our knowledge of the molecular mechanisms of BCG-immunotherapy. J Clin Microbiol, 2002 Sep, 40(9), 3482 - 4 Are three sputum acid-fast bacillus smears necessary for discontinuing tuberculosis isolation? Mathew P, Kuo YH, Vazirani B, Eng RH, Weinstein MP. To evaluate the efficacy of three sputum acid-fast bacillus (AFB) smears to rule out pulmonary tuberculosis, sputum AFB smear and culture results were analyzed at two university-affiliated teaching hospitals . The negative predictive value of the smear increased by only 0.2% on days 2 and 3 each, indicating that in low-prevalence populations, there is limited value in requiring three negative sputum AFB smears before discontinuing tuberculosis isolation. J Clin Microbiol, 2002 Sep, 40(9), 3219 - 22 Comparison of C(18)-carboxypropylbetaine and standard N-acetyl-L-cysteine-NaOH processing of respiratory specimens for increasing tuberculosis smear sensitivity in Brazil; Scott CP et al.; Techniques to improve the sensitivity of smear microscopy would facilitate early tuberculosis (TB) diagnosis and disease control, especially in low-income countries where the positive predictive value is high . C(18)-carboxypropylbetaine (CB-18) is a zwitterionic detergent that helps to compensate for the innate buoyancy of mycobacteria, potentially enhancing recovery by centrifugation . Previous data suggest that CB-18 may increase the sensitivity of smear, culture, and molecular amplification diagnostic testing . The goal of the present study was to evaluate if the sensitivity of the smear technique using light microscopy could be improved by treating respiratory samples with CB-18 . In the first phase, respiratory specimens were collected consecutively from patients with suspected pulmonary tuberculosis in a tertiary-care hospital in Rio de Janeiro, Brazil (236 specimens were analyzed) . After protocol modifications, another 120 respiratory specimens were evaluated . The standard technique was N-acetyl-L-cysteine with sodium hydroxide (NALC-NaOH) treatment, smear concentration with centrifugation, and Ziehl-Neelsen staining . Culture on Lowenstein-Jensen slants was performed on all specimens for use as the "gold standard." No specimens from patients undergoing active TB treatment were included . The initial protocol for CB-18 processing resulted in a sensitivity of 59.6% and specificity of 96.8% compared to standard processing with a sensitivity of 66.0% and specificity of 96.8% . Using the modified protocol, the sensitivity of CB-18 increased to 71.4% with a specificity of 97.0% versus standard processing with a sensitivity of 61.9% and a specificity of 99.0% . The diagnostic yield of acid-fast bacillus smear with CB-18 in the absence of fluorescence microscopy and PCR compared to standard processing with NALC-NaOH was not significantly different, although the power to detect a difference by the modified assay was low. Thorax, 2002 Sep, 57(9), 804 - 9 A meta-analysis of the effect of Bacille Calmette Guérin vaccination on tuberculin skin test measurements; Wang L et al.; BACKGROUND: The accurate diagnosis of latent tuberculosis infection (LTBI) is an important component of any tuberculosis control programme and depends largely on tuberculin skin testing . The appropriate interpretation of skin test results requires knowledge of the possible confounding factors such as previous BCG vaccination . Uncertainty about the effect of BCG vaccination on tuberculin skin testing and the strength with which recommendations are made to individual patients regarding treatment of LTBI have identified a need to analyse the available data on the effect of BCG on skin testing . A meta-analysis of the evidence for the effect of BCG vaccination on tuberculin skin testing in subjects without active tuberculosis was therefore performed . METHODS: Medline was searched for English language articles published from 1966 to 1999 using the key words "BCG vaccine", "tuberculin test/PPD", and "skin testing" . Bibliographies of relevant articles were reviewed for additional studies that may have been missed in the Medline search . Articles were considered for inclusion in the meta-analysis if they had recorded tuberculin skin test results in subjects who had received BCG vaccination more than 5 years previously and had a concurrent control group . Only prospective studies were considered . The geographical location, number of participants, type of BCG vaccine used, type of tuberculin skin test performed, and the results of the tuberculin skin test were extracted . RESULTS: The abstracts and titles of 980 articles were identified, 370 full text articles were reviewed, and 26 articles were included in the final analysis . Patients who had received BCG vaccination were more likely to have a positive skin test (5 TU PPD: relative risk (RR) 2.12 (95% confidence interval (CI)1.50 to 3.00); 2 TU RT23: 2.65 {corrected} (95% CI 1.83 to 3.85) . The effect of BCG vaccination on PPD skin test results was less after 15 years . Positive skin tests with indurations of >15 mm are more likely to be the result of tuberculous infection than of BCG vaccination . CONCLUSIONS: In subjects without active tuberculosis, immunisation with BCG significantly increases the likelihood of a positive tuberculin skin test . The interpretation of the skin test therefore needs to be made in the individual clinical context and with evaluation of other risk factors for infection . The size of the induration should also be considered when making recommendations for treatment of latent infection. Appl Environ Microbiol, 2002 Sep, 68(9), 4583 - 92 Interaction of gene-cloned and insect cell-expressed aminopeptidase N of Spodoptera litura with insecticidal crystal protein Cry1C; Agrawal N et al.; Insecticidal toxins produced by Bacillus thuringiensis interact with specific receptors located in the midguts of susceptible larvae, and the interaction is followed by a series of biochemical events that lead to the death of the insect . In order to elucidate the mechanism of action of B . thuringiensis toxins, receptor protein-encoding genes from many insect species have been cloned and characterized . In this paper we report the cloning, expression, and characterization of Cry toxin-interacting aminopeptidase N (APN) isolated from the midgut of a polyphagous pest, Spodoptera litura . The S . litura APN cDNA was expressed in the Sf21 insect cell line by using a baculovirus expression system . Immunofluorescence staining of the cells revealed that the expressed APN was located at the surface of Sf21 cells . Treatment of Sf21 cells expressing S . litura APN with phosphatidylinositol-specific phospholipase C demonstrated that the APN was anchored in the membrane by a glycosylphosphatidylinositol moiety . Interaction of the expressed receptor with different Cry toxins was examined by immunofluorescence toxin binding studies and ligand blot and immunoprecipitation analyses . By these experiments we showed that the bioactive toxin, Cry1C, binds to the recombinant APN, while the nonbioactive toxin, Cry1Ac, showed no interaction. Appl Environ Microbiol, 2002 Sep, 68(9), 4216 - 24 Genetic heterogeneity in Bacillus sporothermodurans as demonstrated by ribotyping and repetitive extragenic palindromic-PCR fingerprinting; Guillaume-Gentil O et al.; Thirty-eight strains of Bacillus sporothermodurans isolated from ultra-high-temperature (UHT)-treated milk or sterilized milk (UHT isolates) and from animal feed or raw milk (farm isolates) were characterized by automated ribotyping and by repetitive extragenic palindromic (REP)-PCR fingerprinting . By investigating the genetic relationships among isolates from these various sources, the relative importance of different contamination sources could be evaluated . The results of the separate clustering analyses of the PvuII and EcoRI ribopatterns and the REP-PCR patterns were largely consistent with each other and revealed the existence of two main clusters; there was one homogeneous group containing all (REP-PCR) or most (ribotyping) of the UHT isolates, and there was a second more diverse group comprising the farm isolates . A combined three-dimensional analysis of all data showed that three German UHT isolates did not belong to the compact group containing the majority of the UHT isolates . These results demonstrate that B . sporothermodurans is more heterogeneous than previously assumed and that most of the UHT isolates form a genetically distinct subgroup and are capable of producing highly heat-resistant spores . The close genetic relationship of these UHT isolates suggests a clonal origin of a few predominant strains of B . sporothermodurans that can be found in UHT-treated or sterilized milk products. Appl Environ Microbiol, 2002 Sep, 68(9), 4182 - 6 Correspondence of high levels of beta-exotoxin I and the presence of cry1B in Bacillus thuringiensis; Espinasse S et al.; Examination of 640 natural isolates of Bacillus thuringiensis showed that the 58 strains (9%) whose supernatants were toxic to Anthonomus grandis (Coleoptera: Curculionidae) produced between 10 and 175 micro g of beta-exotoxin I per ml . We also found that 55 (46%) of a sample of 118 strains whose culture supernatants were not toxic to A . grandis nevertheless produced between 2 and 5 micro g/ml . However, these amounts of beta-exotoxin I were below the threshold for detectable toxicity against this insect species . Secretion of large amounts of beta-exotoxin I was strongly associated with the presence of cry1B and vip2 genes in the 640 natural B . thuringiensis isolates studied . We concluded that strains carrying cry1B and vip2 genes also possess, on the same plasmid, genetic determinants necessary to promote high levels of production of beta-exotoxin I. J Infect Dis, 2002 Sep 15, 186(6), 807 - 14 Epub 2002 Aug 12. Kinetics of delayed-type hypersensitivity to tuberculin induced by bacille Calmette-Guérin vaccination in northern Malawi; Floyd S et al.; During 1986-1989, a bacille Calmette-Guerin (BCG) vaccine trial was carried out in northern Malawi . The effects of age, sex, and prevaccination delayed-type hypersensitivity (DTH) on the time course of the DTH response over 1-36 months after vaccination were studied in 2418 persons . DTH response increased rapidly, to peak at 31-90 days after vaccination, when most persons had a measurable response . This was followed by a marked decline by 181-365 days, particularly in those <15 years old at vaccination, followed by a more gradual decline . Prevaccination DTH was the single best predictor of postvaccination DTH . BCG-induced DTH responsiveness appears to decline more rapidly in tropical than in temperate environments . This may reflect high prevalence of exposure to other infections, which induce a Th2 bias or compete for "space" within the T lymphocyte compartment . The inability of some persons to mount a persistent DTH response probably reflects genetic background and/or environmental exposure history. EMBO J, 2002 Sep 2, 21(17), 4550 - 9 A cell-cell signaling peptide activates the PlcR virulence regulon in bacteria of the Bacillus cereus group; Slamti L et al.; PlcR is a pleiotropic regulator that activates the expression of genes encoding various virulence factors, such as phospholipases C, proteases and hemolysins, in Bacillus thuringiensis and Bacillus cereus . Here we show that the activation mechanism is under the control of a small peptide: PapR . The papR gene belongs to the PlcR regulon and is located 70 bp downstream from plcR . It encodes a 48-amino-acid peptide . Disruption of the papR gene abolished expression of the PlcR regulon, resulting in a large decrease in hemolysis and virulence in insect larvae . We demonstrated that the PapR polypeptide was secreted, then reimported via the oligopeptide permease Opp . Once inside the cell, a processed form of PapR, presumably a pentapeptide, activated the PlcR regulon by allowing PlcR to bind to its DNA target . This activating mechanism was found to be strain specific, with this specificity determined by the first residue of the penta peptide. Clin Exp Immunol, 2002 Sep, 129(3), 420 - 8 Inhibition of human natural killer cell activity by cereulide, an emetic toxin from Bacillus cereus; Paananen A et al.; The lipophilic toxin, cereulide, emitted by emetic food poisoning causing strains of Bacillus cereus, is a powerful mitochondria toxin . It is highly lipophilic and rapidly absorbed from the gut into the bloodstream . We tested how this toxin influences natural killer (NK) cells, which are important effectors in defence against infections and malignancy . Cereulide inhibited cytotoxicity and cytokine production of natural killer cells, caused swelling of natural killer cell mitochondria, and eventually induced natural killer cell apoptosis . The suppressive effect on cytotoxicity was fast and toxic concentration low, 20-30 microg/l . As the emesis causing concentration of cereulide is around 10 microg/kg of total body mass, our results suggest that emesis causing or even lower doses of cereulide may also have a systemic natural killer cell suppressive effect. Biochem Soc Trans, 2002 Aug, 30(4), 610 - 3 Production of cobalamin and sirohaem in Bacillus megaterium: an investigation into the role of the branchpoint chelatases sirohydrochlorin ferrochelatase (SirB) and sirohydrochlorin cobalt chelatase (CbiX); Leech HK et al.; One of the four operons required for cobalamin biosynthesis in Bacillus megaterium is also associated with sirohaem synthesis, and contains three genes, sirA, sirB and sirC . By undertaking functional complementation experiments and in vitro assays using recombinantly produced enzymes, we have been able to demonstrate that (1) SirA acts as a uroporphyrinogen III methyltransferase, transforming uroporphyrinogen III into precorrin-2, (2) SirC acts as an NAD(+) dehydrogenase, responsible for the oxidation of precorrin-2 into sirohydrochlorin, and (3) SirB acts as a ferrochelatase, responsible for the insertion of a ferrous ion into sirohydrochlorin to give sirohaem . Comparative sequence analysis reveals that the primary structure of SirB is highly similar to that of the cobalt chelatase involved in cobalamin biosynthesis in Bacillus megaterium, CbiX, with the exception that CbiX contains a C-terminal histidine-rich motif . Surprisingly, CbiX has been shown (using EPR) to contain a 4Fe-4S centre, a redox centre that is absent from SirB. Biochem Soc Trans, 2002 Aug, 30(4), 595 - 600 Structural diversity in metal ion chelation and the structure of uroporphyrinogen III synthase; Schubert HL et al.; All tetrapyrroles are synthesized through a branched pathway, and although each tetrapyrrole receives unique modifications around the ring periphery, they all share the unifying feature of a central metal ion . Each pathway maintains a unique metal ion chelatase, and several tertiary structures have been determined, including those of the protoporphyrin ferrochelatase from both human and Bacillus subtilus, and the cobalt chelatase CbiK . These enzymes exhibit strong structural similarity and appear to function by a similar mechanism . Met8p, from Saccharomyces cerevisiae, catalyses ferrochelation during the synthesis of sirohaem, and the structure reveals a novel chelatase architecture whereby both ferrochelation and NAD(+)-dependent dehydrogenation take place in a single bifunctional active site . Asp-141 appears to participate in both catalytic reactions . The final common biosynthetic step in tetrapyrrole biosynthesis is the generation of uroporphyrinogen by uroporphyrinogen III synthase, whereby the D ring of hydroxymethylbilane is flipped during ring closure to generate the asymmetrical structure of uroporphyrinogen III . The recently derived structure of uroporphyrinogen III synthase reveals a bi-lobed structure in which the active site lies between the domains. J Infect Dis, 2002 Sep 1, 186(5), 678 - 83 Epub 2002 Aug 05. Detection of antibody to Mycobacterium tuberculosis protein antigens in the cerebrospinal fluid of patients with tuberculous meningitis; Chandramuki A et al.; Antibodies against Mycobacterium tuberculosis antigens were detected by enzyme-linked immunosorbent assay in cerebrospinal fluid (CSF) samples obtained from 442 patients with tuberculous meningitis (TBM) and 102 control patients . Antibodies were found in the CSF of 87% of patients with clinical (culture-negative) TBM, 72% of patients with culture-positive TBM, and 65% of patients with autopsy-proven TBM . That anti-M . tuberculosis antibodies were detected in the CSF of patients with clinically diagnosed cases more frequently than in patients with culture-positive cases suggests that the detection of antibodies in CSF tends to decrease as bacillary load increases . Of the patients with clinical TBM who were coinfected with human immunodeficiency virus (HIV), 70% exhibited anti-M . tuberculosis antibody in CSF, which suggests that antibody responses in this group were substantially weaker than those in HIV-negative patients with clinical TBM . Some groups showed a stronger response to certain antigens, which suggests that antigen recognition patterns may be specific for the stage of disease. Curr Treat Options Oncol, 2002 Oct, 3(5), 403 - 11 Superficial bladder cancer; Lamm DL; Superficial bladder cancer can be resected with minimal morbidity, but the patients are at high risk for tumor recurrence . Tumors can be divided into low-, intermediate-, and high-risk categories based on tumor grade, stage, and pattern of recurrence . Low-risk tumors are best treated with a single instillation of chemotherapy such as thiotepa, doxorubicin, or mitomycin . Intermediate-risk tumors can be treated with chemotherapy, but, similar to high-risk tumors, will often require immunotherapy . High-risk tumors are best treated with intravesical bacille Calmette-Guerin (BCG) using a 3-week maintenance schedule . Side effects of BCG immunotherapy can be reduced by logarithmic reductions in the dosage of BCG . Patients who fail BCG may be rescued with BCG plus interferon-alfa or radical cystectomy. Pest Manag Sci, 2002 Aug, 58(8), 791 - 8 High level methoprene resistance in the mosquito Ochlerotatus nigromaculis (Ludlow) in central California; Cornel AJ et al.; In the summer of 1998, failures of methoprene field applications to control the mosquito Ochlerotatus nigromaculis (Ludlow) were noticed in several pastures in the outskirts of Fresno, California, USA . Effective control with methoprene had been achieved for over 20 years prior to this discovery . Susceptibility tests indicated that the Fresno Oc nigromaculis populations had developed several thousand-fold higher LC50 and LC90 tolerance levels to methoprene compared with methoprene-naive populations . The synergists piperonyl butoxide (PBO), S,S,S-tributyl phosphorotrithioate and 3-octylthio-1,1,1-trifluoro-2-propanone had little synergistic effect, suggesting that the mechanism of methoprene tolerance was not mediated by P450 monooxygenase or carboxylesterase enzyme degradation . As part of initiating a resistance management strategy, partial reversion back to methoprene susceptibility was achieved in a resistant population after six consecutive applications of Bacillus thuringiensis israelensis Goldberg & Marga coupled with two oil and two pyrethrum + PBO applications. Sheng Wu Gong Cheng Xue Bao, 2002 May, 18(3), 335 - 8 {A novel resolution vector with Bacillus thuringiensis plasmid replicon ori44}; Wu L et al.; The resolution recognization sites of transposon Tn4430 of Bacillus thuringiensis was inserted into cloning vector pRSET B and pUC19, resulting recombinant plasmids pBMB1201 and pBMB1202 . Both of the mini res fragments, BamHI/HindIII fragment in pBMB1201 and EcoRI/HindIII fragment in pBMB1202, were ligated to the 3.3 kb EcoRI/HindIII fragment of shuttle vector pHT3101, which contained the ori . Ec, ampr and emr antibiotic resistant genes, resulting recombinant plasmid pBMB1203 . After deleted the BamHI and EcoRI sites which located ouside the two res sites, resolution vector pBMB1204 was resulted . There are multiple cloning sites between two copies of resolution sites which have the same direction . The plasmid replication origin ori44, which come from B . thuringiensis sub sp . kurstaki strain YBT-1520, was inserted into the multiple cloning sites of pBMB1204 and then resolution shuttle vector pBMB1205 was obtained . With spectinomycin resistant gene as target, it was found that the resolution rate is 100% and the stability of the resolved plasmid is 93% . Using this shuttle vector, antibiotic resistance markers and other non-B . thuringiensis DNA can be selectively eliminated after the selection of transformants by antibiotic resistance marker . This vector is very useful to solve the gene safety problem while has no effect on target gene expression. Curr Microbiol, 2002 Oct, 45(4), 299 - 302 Isolation and characterization of a strain of Bacillus thuringiensis ssp . kurstaki containing a new delta-endotoxin gene; Li MS et al.; A strain of Bacillus thuringiensis that showed significantly high toxicity to Plutella xylostella and Spodoptera exigua was isolated from a Korean soil sample and characterized . The isolate, named B . thuringiensis K1, was determined to belong to ssp . kurstaki (H3a3b3c) type by an H antisera agglutination test and produced bipyramidal inclusions . Plasmid pattern of K1 was different from that of the reference strain, ssp . kurstaki HD-1, but the parasporal inclusion protein profile of K1 had two major bands that were similar in size to those of ssp . kurstaki HD-1 . To verify the delta-endotoxin gene types of K1, PCR analysis with specific cry gene primers was performed to show that K1 contained a new cry gene in addition to cry1Aa, cry1Ab, cry1Ac, cry1E and cry2 genes . PCR-amplified region of the new cry gene, cryX, showed 79% similarity to cry1Fa1 gene (GenBank Accession No . M63897) . In an insect toxicity assay, K1 had higher toxicity against Plutella xylostella and S . exigua than ssp . kurstaki HD-1. Curr Microbiol, 2002 Oct, 45(4), 265 - 71 Conversion of fatty acids by Bacillus sphaericus-like organisms; Kuo TM et al.; Bacillus sphaericus species are mesophilic round-spored organisms that readily utilize fatty acid-based surfactants during growth, but their ability to modify fatty acids is unknown . Among 57 B . sphaericus-like strains tested for fatty acid transformation activity in Wallen fermentation (WF) medium, ten converted oleic acid to a new product determined by gas chromatography - mass spectrometry (GC-MS) to be 10-ketostearic acid (10-KSA) . Additionally, a few other strains converted ricinoleic acid and linoleic acid to new products that remain to be characterized . Unlike most microbial hydrations of oleic acid, which produce a mixture of 10-KSA and 10-hydroxystearic acid, the conversion of oleic acid by B . sphaericus strains was unique in that 10-KSA was the sole reaction product . By replacing dextrose with sodium pyruvate in WF and adjusting to pH 6.5, conversion of oleic acid to 10-KSA by strain NRRL NRS-732 was improved from about 11% to more than 60% . Using the defined optimal conditions, the conversion reaction was scaled up in a stirred-batch reactor by using technical-grade oleic acid as substrate . This is the first report on the characterization of fatty acid conversions by B . sphaericus species. Mikrobiol Z, 2002 Mar-Apr, 64(2), 44 - 8 {Influence of composition of the nutrient medium on growth and development of entomopathogenic bacteria Bacillus thuringiensis}; Dregval' OA et al.; New medium for cultivation of Bacillus thuringiensis has been elaborated with the help of a simplex method for mathematical planning the experiment . The optimized medium containing green syrup, corn extract, sodium humate and mineral salts, allowed to increase the biomass titer by 45%, endotoxin by 220% as compared to the initial medium . Insecticide activity of Bacillus thuringiensis culture grown on the new culture medium exceeds considerably that on the initial culture medium. J Biochem Mol Biol Biophys, 2002 Aug, 6(4), 279 - 82 Biochemical characteristics of chitinase enzyme from Bacillus sp . of Kamojang Crater, Indonesia; Natsir H et al.; Chitinase and chitindeacetylase are enzymes capable of degrading chitin into chitooligomers and chitosan . The chitinases characterized and purified in this study were extracted from the acidophillic Bacillus sp . isolated from Kamojang Crater West Java Indonesia . When grown in liquid media containing colloidal chitin, the optimum chitinase activity of the acidophilic isolate was reached after 4-5 days of incubation . The optimum temperature and pH of the chitinase and chitin deacetylase were found at 37 degrees C and pH 5 . When incubated at pH 5, the activity of chitin deacetylase was increased; after 3 h, the activity was 1.5 times of the control . The enzyme was stable at pH 4, after 2 h incubation, the activity was still 80% of the control . The chitinase and chitin deacetylase activities were not influenced by Mg(++) nor Ca(++), Ni(++) and Cu(++) inhibited the chitinase activity, while chitin deacetylase activity was not affected by Cu(++) addition . When 1 mM of EDTA was added, the enzyme activity was reduced 40 to 50%. Postgrad Med J, 2002 Aug, 78(922), 449 - 54 Use of bacille Calmette-Guérin in superficial bladder cancer; Meyer JP et al.; Intravesical bacille Calmette-Guerin (BCG) has been used by urologists for several years after its first reported use as a cancer therapy in the 1930s . Morales in 1976 described the usage of BCG as a once weekly intravesical instillation for six weeks; this is a treatment regimen that still exists today . Its success as a treatment depends on it being used appropriately . It is employed: (1) to treat carcinoma in situ or occasionally residual papillary tumours; (2) to reduce the number and frequency of recurrent high grade superficial tumours; and (3) to prevent disease progression (although this remains a controversial point, on which there is no consensus view) . Unfortunately, the more widespread use of BCG is often limited due its high side effect profile . Present research is directed towards reducing its side effect profile, improving its efficacy, and understanding its exact mechanism of action, which is not fully understood. Protein Expr Purif, 2002 Aug, 25(3), 426 - 9 A purification method of the diagnostic enzyme Bacillus uricase using magnetic beads and non-specific protease; Nishiya Y et al.; A simple purification method of the Bacillus uricase (Uao) was newly developed . The gene coding for Uao with a C-terminal 6-histidine tag (Uao-HT) was constructed and overexpressed . Using the non-specific proteases, such as proteinase K, the tag was easily removed because Uao-HT includes its C-terminal region to be specifically cleaved by them . Such treatment of Uao-HT with the proteases did not affect its enzymatic properties and enabled us to purify it from the crude extract with a single-step protocol; the cell lysate containing Uao-HT was mixed with the Ni ion-chelating magnetic beads and then the adsorbed enzyme was eluted with the proteinase K-containing buffer after untagged proteins were washed out . The isolated enzyme yielded a single band on SDS-PAGE and was fully active . This method is extremely useful for high-throughput purification of mutants because of compatibility with automation. TB HIV . 1996 Jun-Aug;(11):26. Tuberculosis and HIV infection in Brazil -- update and overview; Dalcolmo M; PIP: In Brazil, tuberculosis (TB) has become an important public health problem with approximately 90,000 cases reported annually . Compounding the TB problem is the interaction of the growing HIV/AIDS epidemic with TB, with potential implications for intranosocomial transmission to immunodeficient patients and health care workers, as well as for the emergence of multidrug-resistant strains . The increased number of reported TB cases is attributed to the deterioration in socioeconomic conditions, the decentralization and restructuring of local TB programs, and the HIV epidemic . TB is the third most common AIDS-defining event; hence, increase in TB cases could be attributed to coinfection with HIV . To this effect, the Tuberculosis and AIDS Programs created a joint committee in order to define national guidelines for the prevention, case management, and treatment of HIV-associated TB . These guidelines recommend 1) preventive therapy with Isoniazid (300 mg daily for 6 months); and 2) Rifampicin-Isoniazid-Pyrazinamide daily for 2 months plus Rifampicin-Isoniazid for the following 7 months for those with active TB . In addition to these strategies, Bacillus-Calmette Guerin vaccination is mandatory for newborns and children under age 4 years, especially for all HIV positive asymptomatic newborns of HIV-infected or AIDS mothers . Foro Mund Salud, 1990, 11(2), 173 - 8 {Monetary incentives for a more effective immunization program}; Zi W; PIP: In 1984 an insurance plan for child immunization was introduced in the counties of Wuji, Linzhang, an Zunhua, and in the city of Xingtai in the province of Hebei, China . The remuneration of village physicians and payment for vaccination services was linked to their effectiveness . In Wuji county children under 2 up to the age of 7 could be enrolled . If a child contracted measles, $8.50 was paid as compensation, $29 for tetanus, $43 for diphtheria, and $57 for poliomyelitis . If death was caused by one of these diseases, $85 was paid . 84% of children of this age range participated in the plan . 36% of the money from policy purchases of $36,000 was allocated to compensation and administration and 64% to municipal health centers for replacement of supplies and to village doctors performing vaccinations whose annual income ranged between $86 and $286 plus $37 for vaccinations . In the 1st year 31 cases were compensated for a total of $600 . In the other countries similar schemes were operational: rural doctors pitched in with 5-20-25% of compensation, and the health care system paid the rest . In 1987, two-thirds of cities in the province adopted this plan enlisting 2,559,780 children (31% of those under 7) and accumulating a total fund of $2,500,000 . A 1987 sample of 36, 992 children indicated a 94% coverage for BCG (bacillus Calmette-Guerin), 85% for poliomyelitis, 80% for DPT (diphtheria-pertussis-tetanus), 80% for measles, 90% for Japanese B encephalitis and epidemic meningitis . In 1986, there were 273,000 fewer cases of measles, polio, diphtheria, pertussis, epidemic meningitis, and Japanese encephalitis with 4200 fewer deaths, and 3000 fewer incapacities (or possibly 1,000,000, 15,000 and 5000 fewer, respectively, because of nonreporting) . Measles morbidity declined from 16/100,000 in 1986 to 3.7/100,000 in the first 11 months of 1987 . Curr Microbiol, 2002 Sep, 45(3), 221 - 5 Detection of enterotoxin genes in mosquito-larvicidal Bacillus species; Yuan Z et al.; The presence of the B component of hemolysin BL (hblA), enterotoxin BceT (bceT), and enterotoxin S (entS) genes in mosquito-larvicidal Bacillus sp., including 25 B . sphaericus and 4 B . thuringiensis subsp . israelensis strains, has been analyzed by multiplex PCR in this study . The results showed that all four B . thuringiensis strains contain the hblA gene and the sequences of bceT and entS genes . However, none of the enterotoxin gene sequences were detected in the B . sphaericus strains . The enterotoxin production in all strains has also been analyzed by using two commercial immunoassay kits (TECRA and RPLA), and it has been proved that all the B . thuringiensis strains and one B . cereus strain can produce enterotoxins during growth . No enterotoxin activity could be detected in B . sphaericus strains. Curr Microbiol, 2002 Sep, 45(3), 217 - 20 Protection from UV-B damage of mosquito larvicidal toxins from Bacillus thuringiensis subsp . israelensis expressed in Anabaena PCC 7120; Manasherob R et al.; A transgenic strain of the nitrogen-fixing filamentous cyanobacterium Anabaena PCC 7120 protected expressed delta-endotoxin proteins of Bacillus thuringiensis subsp . israelensis from damage inflicted by UV-B, a sunlight component that penetrates Earth's ozone layer . This organism, which serves as a food source to mosquito larvae and could multiply in their breeding sites, may solve the environment-imposed limitations of B . thuringiensis subsp . israelensis as a mosquito biological control agent. Curr Microbiol, 2002 Sep, 45(3), 187 - 90 Expression of Bacillus stearothermophilus LV cadmium resistance genes in Escherichia coli causes hypersensitivity to cadmium chloride; Nerey Md Mdel C et al.; Determination of the nucleotide sequence of a 4.5-kb chromosomal DNA fragment of Bacillus stearothermophilus LV revealed two open reading frames (ORFs) of 121 and 727 amino acids (aa) that exhibit a high degree of similarity with the cadC and cadA cadmium resistance genes of a number of microorganisms . Transfer and expression of the B . stearothermophilus LV cadA or cadC/ cadA genes in E . coli caused increased cadmium chloride susceptibility in the bacterial host . Transfer of cadC alone did not result in any detectable phenotypic change in E . coli. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1998, 30(2), 107 - 113 Cloning and Expression of the pga Gene of Bacillus megaterium in E . coli; Huang YH et al.; The pga gene was amplified by PCR from Bacillus megaterium CA4098, cloned into pKK223-3 vector and expressed in E . coli HB101 . The pga sequence was also determined . The deduced amino acids sequence was compared with those of PGAs from other bacteria and showed that they were similar, especially the active sites were strongly conserved. J Appl Microbiol, 2002, 93(3), 374 - 9 Chitinolytic activities in Bacillus thuringiensis and their synergistic effects on larvicidal activity; Liu M et al.; AIMS: To investigate the distribution of chitinase in Bacillus thuringiensis strains, and the enhancing effects of the chitinase-producing B . thuringiensis strains on insecticidal toxicity of active B . thuringiensis strain against Spodoptera exigua larvae . METHODS AND RESULTS: The chitinolytic activities of B.thuringiensis strains representing the 70 serotypes were investigated by the whitish opaque halo and the colorimetric method . Thirty-eight strains produced different levels of chitinase at pH 7.0, and so did 17 strains at pH 10.0 . The strain T04A001 exhibited the highest production, reaching a specific activity of 355 U ml(-1) in liquid medium . SDS-PAGE and Western blotting showed that the chitinase produced by some B . thuringiensis strains had a molecular weight of about 61 kDa . The bioassay results indicated that the chitinase-producing B . thuringiensis strains could enhance the insecticidal activity of B . thuringiensis strain DL5789 against S . exigua larvae, with an enhancing ratio of 2.35-fold . CONCLUSION: This study demonstrated that chitinase was widely produced in B . thuringiensis strains and some of the strains could enhance the toxicity of active B . thuringiensis strain . SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first investigation devoted exclusively to analyse the distribution of chitinase in B . thuringiensis . It infers that the chitinase produced by B . thuringiensis might play a role in the activity of the biopesticide. Biochemistry, 2002 Aug 20, 41(33), 10446 - 53 Identification of key amino acid residues in the assembly of enzymes into the pyruvate dehydrogenase complex of Bacillus stearothermophilus: a kinetic and thermodynamic analysis; Jung HI et al.; Structural studies have shown that electrostatic interactions play a major part in the binding of dihydrolipoyl dehydrogenase (E3) to the peripheral subunit-binding domain (PSBD) of the dihydrolipoyl acyltransferase (E2) in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus . The binding is characterized by a small, unfavorable enthalpy change (deltaH degrees = +2.2 kcal/mol) and a large, positive entropy change (TdeltaS degrees = +14.8 kcal/mol) . The contributions of individual surface residues of the PSBD of E2 to its interaction with E3 have been assessed by alanine-scanning mutagenesis, surface plasmon resonance detection, and isothermal titration calorimetry . The mutation R135A in the PSBD gave rise to a significant decrease (120-fold) in the binding affinity; two other mutations (R139A and R156A) were associated with smaller effects . The binding of the R135A mutant to E3 was accompanied by a favorable enthalpy (deltaH degrees = -2.6 kcal/mol) and a less positive entropy change (TdeltaS degrees = +7.2 kcal/mol) . The midpoint melting temperature (T(m)) of E3-PSBD complexes was determined by differential scanning calorimetry . The R135A mutation caused a significant decrease (5 degrees C) in the T(m), compared with the wild-type complex . The results reveal the importance of Arg135 of the PSBD as a key residue in the molecular recognition of E3 by E2, and as a major participant in the overall entropy-driven binding process . Further, the effects of mutagenesis on the deltaCp of subunit association illustrate the difficulties in attributing changes in heat capacity to specific classes of interactions. J Commun Dis, 2001 Jun, 33(2), 116 - 20 Comparative toxicity of selected larvicidal formulations against Anopheles stephensi Liston and Aedes aegypti Linn; Mittal PK et al.; Toxicity of selected larvicidal formulations of Bacillus sphaericus, Bacillus thuringiensis H-14 israelensis (Bti) and insect growth regulators was determined against Anopheles stephensi and Aedes aegypti, the two major urban mosquito vectors of malaria and dengue fever in India respectively . The study revealed that the formulations of Bacillus thuringiensis H-14 and IGR compounds were highly toxic against both the mosquito species but Bacillus sphaericus formulation was less toxic against Aedes aegypti and has a limited potential for use against Aedes aegypti . Of the two Bti formulations, 'Vectobac' formulation was more toxic against both the species than 'BMP-1442X' formulation . Bti formulations were more toxic against Aedes aegypti (LC50 = 0.06 and 0.14 mg/Litre) than against An . stephensi (LC50 = 0.14 and 0.81 mg/Litre), while B . sphaericus formulation was more toxic against An . stephensi LC50 = 0.031 mg/Litre than Aedes aegypti LC50 = 0.294 mg/Litre . Among different larvicidal formulations tested in this study, IGR compounds were found to be toxic against both the mosquito species at very low concentrations (EC50 values ranging between 0.0001 and 0.0004 ppm). Contraception, 2002 Jul, 66(1), 47 - 56 Evaluation of interaction potential of certain concurrently administered drugs with pharmacological and pharmacokinetic profile of centchroman in rats; Khurana M et al.; Centchroman (Ormeloxifene) is a nonsteroidal, selective estrogen receptor modulator, oral contraceptive and anticancer agent, and is intended for long-term use by women . In view of its vast clinical application and the interaction of steroidal oral contraceptives with certain commonly used therapeutic agents, evaluation of interaction of certain concomitantly administered therapeutic agents (ibuprofen, rifampicin, diazepam, salbutamol, nifedipine, paracetamol, haloperidol, and tetracycline), in terms of both the postcoital contraceptive efficacy and pharmacokinetic profile, with centchroman was undertaken in female Sprague-Dawley rats . Among the representatives from each commonly used therapeutic category, interaction (pharmacokinetic) was observed with ibuprofen (60 mg/kg, twice daily), haloperidol (0.7 mg/kg, twice daily), and tetracycline (140 mg/kg, twice daily) coadministration on Days 1 through 5 postcoitum . Of these three therapeutic agents, only tetracycline interfered with the contraceptive efficacy of centchroman . It reduced the bioavailability of centchroman and its active metabolite by increasing their excretion through bile and feces . Increased metabolite excretion on tetracycline coadministration indicates the enterohepatic recirculation of the metabolite, not the parent drug . However, the effect of tetracycline was negated by the inclusion of lactic acid bacillus spores in the regimen. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1998, 30(4), 352 - 356 High Efficient Expression in Escherichia coli of Chitinase Gene Cloned from Bacillus circulans C-2; Wang YL et al.; Subcloning analysis of a cloned DNA fragment from Bacillus circulans containing the chitinase gene Chi1 showed that the chitinase gene lies on a 1.7kb PstI-StyI fragment . The chitinase gene could be expressed in Escherichia coli strains JM107, DH5alpha, XL1-blue and TG-1 with various efficiencies . The expression level of chitinase gene was highest in JM107, which was almost the same as that in B . circulans C-2 . The molecular weight of extracellular chitinase was 66 kD by SDS-PAGE analysis . Cell location determination of the expressed chitinase showed that the enzyme existed not only in cell periplasm and cytoplasm, but also in extracellular broth . When the expression of the enzyme was optimal, the distribution of enzyme activity in extracellular broth, periplasm and cytoplasm was 35.8%, 32.1% and 32.9%, respectively. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1998, 30(4), 325 - 330 Expression of Recombinant alpha-Acetolactate Decarboxylase and Its Characterization; Yin D et al.; alpha-Acetolactate decarboxylase(alpha-ALDC)gene has been cloned from Baillus brevis using PCR amplification . The amplified 0.97 kb DNA fragment was confirmed to be known alpha- ALDC gene by DNA sequencing . The fragment was inserted into the vector pBV220 to construct an expression plasmid pBVYI . This recombinant plasmid over expressed alpha-ALDC in E . coli DH5alpha . The alpha-ALDC activity of recombinant bacterium was 10 000-fold higher than that of Bacillus brevis . After purification, the properties of the recombinant alpha-ALDC were studied . The activity of this enzyme could be stimulated by Mn(2+), Sn(2+) and inhibited by Zn(2+), Cd(2+), Fe(2+), Co(2+) and Cu(2+) . Moreover, amino acid modifiers could inhibit differently its activity . The optimum pH of the enzyme reaction was 5.5. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 1998, 30(6), 579 - 584 Overexpression, Purification and Characterization of Thermostable Catechol 2,3-dioxygenase; Zhang W et al.; Catechol 2,3-dioxygenase(EC 1.13.11.2) from Bacillus stearothermophilus has been shown to be highly thermostable . In order to obtain sufficient enzyme for its characterization, and to analyze the molecular basis of its intrinsic stability, the gene coding for this enzyme was sub-cloned and overexpressed in E . coli . After heat denaturation, ammonium sulfate fractionation, DEAE-52 ion-exchange chromatography and phenyl-Sepharose CL-4B hydrophobic chromatography, the enzyme was purified to homogeneity and the yield was 16% . The enzyme is a homotetramer . The molecular weight of each subunit is 36.4 kD as determined by SDS-PAGE . Using the tyrosine difference spectrum method, the extinction coefficient of the enzyme at 279.2 nm is estimated as 89(mmol/L)(-1).cm(-1), the optimum temperature and pH of the enzyme is 60 degrees and 8.0, respectively . The K(m) for catechol of the enzyme is 1.24x10(-5) mol/L, K(cat) is 24 000 min(-1) . Acetone competitively inhibited the enzyme activity to catechol, with an inhibition constant K(i) of about 165 mmol/L . The thermostability of the enzyme is reflected by its unaltered catalytic activity over 1 h at 65 degrees . Irreversible thermal denaturation becomes significant between 70-80 degrees . THE pH stability of the enzyme and its resistance toward chemical denaturation such as urea and SDS confirm the intrinsic stability of the enzyme . It was also studied that the effect of different pHs on the molecular structure of the enzyme, using circular dichroism spectra and intrinsic fluorescence analysis. DNA Cell Biol, 2002 May-Jun, 21(5-6), 367 - 73 Molecular mechanisms of Bacillus endophthalmitis pathogenesis; Callegan MC et al.; Bacillus endophthalmitis is one of the most devastating intraocular infections, frequently resulting in significant vision loss, if not loss of the eye itself, in only a few days . This review summarizes recent research focused on characterizing the interactions between Bacillus and the host response during endophthalmitis . Analyses of the contribution of Bacillus toxins and cell wall components, and the behavior of the organism during progressive disease are discussed . A better understanding of the host/pathogen interactions occurring during endophthalmitis is critical for the development of novel therapeutic agents designed to impede the progression of infection and protect vision. Phytochemistry, 2002 Sep, 61(1), 15 - 24 Long chain polyunsaturated fatty acid production and partitioning to triacylglycerols in four microalgae; Tonon T et al.; Gas chromatographic profiling of fatty acids was performed during the growth cycle of four marine microalgae in order to establish which, if any, of these could act as a reliable source of genes for the metabolic engineering of long chain polyunsaturated fatty acid (LC-PUFA) synthesis in alternative production systems . A high-throughput column based method for extraction of triacylglycerols (TAGs) was used to establish how much and at what stage in the growth phase LC-PUFAs partition to storage lipid in the different species . Differences in the time course of production and incorporation of docosahexaenoic acid (22:6n-3, DHA) and eicosapentaenoic acid (20:5n-3, EPA) into TAGs were found in the marine microalgae Nannochloropsis oculata (Eustigmatophyceae), Phaeodactylum tricornutum and Thalassiosira pseudonana (Bacillariophyceae), and the Haptophyte Pavlova lutheri . Differences were not only observed between species but also during the different phases of growth within a species . A much higher percentage of the total cellular EPA was partitioned to TAGs in stationary phase cells of N . oculata compared to P . tricornutum . Although P . tricornutum produces DHA it does not partition it to TAGs . Both T . pseudonana and P . lutheri produce EPA and DHA and partition these to TAGs during the stationary phase of growth . These two species are therefore good candidates for further biochemical and molecular analysis, in order to understand and manipulate the processes that are responsible for the incorporation of LC-PUFAs into storage oils . Biosci Biotechnol Biochem, 2002 Jun, 66(6), 1270 - 5 Endophytic bacillus sp . isolated from the interior of balloon flower root; Cho SJ et al.; A bacterial strain, designated CY22, was isolated from the interior of balloon flower (Platycodon grandiflorum) root in the Republic of Korea . The isolate coproduced an iturin-like antifungal compound and a surfactin-like potent biosurfactant . Analysis of the 16S-rDNA of strain CY22 showed that the isolate was a member of Bacillus . High similarities were observed between strain CY22 and Bacillus sp . TKSP 24, and between strain CY22 and B . subtilis 168 . Phylogenetic analysis based on 16S-rDNA sequences showed that strain CY22 was closely related to Bacillus sp . The main whole-cell fatty acids were anteiso-C15:0 (37%), C17:0 (5.1%), and iso-C15:0 (27.7%) . DNA G+C content was 54 mol% . Based on phylogenetic inference, phenotypic and chemotaxonomic characteristics, this endophytic strain Bacillus sp . CY22 was assigned to the genus Bacillus. Biosci Biotechnol Biochem, 2002 Jun, 66(6), 1246 - 55 Cloning and structural analysis of bglM gene coding for the fungal cell wall-lytic beta-1,3-glucan-hydrolase BglM of Bacillus circulans IAM1165; Asano T et al.; Bacillus circulans IAM1165 produces isoforms of beta-1,3-glucan-hydrolases . Of these enzymes, the 42-kDa enzyme BgIM degrades Aspergillus oryzae cell walls the most actively . A gene coding for a BgIM precursor consisting of 411 amino acid residues was cloned . The 27 N-terminal amino acid sequence of the precursor is a signal peptide . The 141 C-terminal amino acid sequence showed a motif of carbohydrate-binding module family 13 . This domain bound to pachyman, lichenan, and A . oryzae cell walls . The central domain showed a bacterial beta-1,3-glucan-hydrolase motif belonging to glycosyl hydrolase family 16 . By removal of the C-terminal domain in the IAM1165 culture, mature BglM was processed to several 27-kDa fragments that hydrolyze a soluble beta-1,3-glucan. Biomaterials, 2002 Oct, 23(19), 3957 - 67 Enzymatic synthesis of dextran-containing hydrogels; Ferreira L et al.; Dextran, a natural glucose-containing polysaccharide, has been acylated by Proleather FG-F and lipase AY, a protease and lipase from Bacillus sp . and Candida rugosa, respectively, in anhydrous dimethylsulfoxide in the presence of vinyl acrylate (VA) . The efficiency of the reaction in the presence of Proleather FG-F and the isolated yields were ca . 71% and 45%, respectively . Dextran derivatized with VA (dexT70-VA) was characterized by gel permeation chromatography and its structure was established by NMR indicating two positional isomers at the 2 and 3 positions on the glucose moieties in equal amounts . Furthermore, the dextran glucopyranose residues were mono-substituted . The benefits of the biocatalytic synthesis of dextran acrylates was demonstrated using 4-dimethylaminopyridine as a chemical catalyst . Gels were prepared by free radical polymerization of aqueous solutions of dexT70-VA with different degrees of substitution and monomer concentrations . Intermolecular linkages and physical entanglements are predominantly formed by concentrated solutions, however, a part of the acrylate groups did not react . Gel pore sizes were calculated from swelling experiments and ranged from ca . 18 to 182 A. J Ind Microbiol Biotechnol, 2002 Aug, 29(2), 51 - 4 The radiation resistance of ascospores and sclerotia of Pyronema domesticum; Richter SG et al.; The Food and Drug Administration has become aware of several instances where supposedly sterile medical surgical products made of Chinese cotton have been found to contain viable Pyronema domesticum . The aim of this research was to determine the gamma and electron beam radiation resistance values for the two dormant phases (ascospores and sclerotia) of P . domesticum . The resistance values were obtained by developing a standardized system to cultivate, purify, and harvest biological indicators containing sclerotia or ascospores . Ascospores were more resistant to radiation than sclerotia . The D(10) values for sclerotia were 0.79 and 1.09 kGy for strains 32030 and 14881, respectively . The resistance value for wild type ascospores was 2.83 kGy . The current standard for assuring radiation sterilization of medical devices is ISO 11137 . This standard was developed to address the propensity for highly radiation-resistant organisms such as P . domesticum . Prior to the standard, biological indicators such as Bacillus pumilus, having a nominal D(10) value or 1.7 kGy, were used to determine the sterility of many medical devices. Microbiol Immunol, 2002, 46(6), 365 - 9 Rapid and sensitive detection method of a bacterium by using a GFP reporter phage; Funatsu T et al.; A rapid, sensitive, and convenient method for detecting a specific bacterium was developed by using a GFP phage . Here we describe a model system that utilizes the temperate Escherichia coli-restricted bacteriophage lambda, which was genetically modified to express a reporter gene for GFP to identify the colon bacillus E . coli in the specimen . E . coli infected with GFP phage was detected by GFP fluorescence after 4-6 hr of incubation . The results show that a few bacteria in a specimen can be detected under fluorescence microscopy equipped with a sensitive cooled CCD camera . When E . coli and Mycobacterium smegmatis were mixed in a solution containing GFP phage, only E . coli was infected, indicating the specificity of this method . The method has the following advantages: 1) Bacteria from biological samples need not be purified unless they contain fluorescent impurities; 2) The infection of GFP phage to bacteria is specific; 3) The fluorescence of GFP within infected bacteria enables highly sensitive detection; 4) Exogenous substrates and cofactors are not required for fluorescence . Therefore this method is suitable for any phage-bacterium system when bacteria-specific phages are available. Environ Toxicol Chem, 2002 Aug, 21(8), 1664 - 72 Responses of songbirds to aerial spraying of the microbial insecticide Bacillus thuringiensis var . kurstaki (FORAY 48B) on Vancouver Island, British Columbia, Canada; Sopuck L et al.; Use of bacterial insecticides containing Bacillus thuringiensis var . kurstaki (Btk) is gaining popularity as an environmentally safe control measure against lepidopteran pests, but indirect effects on nontarget organisms through reduced prey base have received little attention . Aerial spraying of Btk (Foray 48B) over a 12,803-ha area on southeastern Vancouver Island (BC, Canada) in May to June 1999 as part of a gypsy moth (Lymantria dispar) control program provided us with an opportunity to examine the responses of songbirds to spray application . To obtain an estimate of species richness and relative abundance, we conducted standard songbird point-count surveys in Garry oak (Quercus garryana)-dominated habitats in Btk-sprayed and unsprayed areas in April to June 1999 and, one year after spraying, in 2000 . These surveys revealed no patterns consistent with adverse effects of spraying on the relative abundance of adult birds or singing males for any of the species, whether examined individually or when combined into foraging guilds . An exception was the spotted towhee (Pipilo maculatus), which in 1999, but not in 2000, occurred at significantly lower numbers in sprayed plots after Btk treatment . Intensive searches of plots in sprayed and unsprayed areas revealed no differences in the numbers of songbird broods between the two areas for any of the species examined. Chemosphere, 2002 Jul, 48(4), 427 - 35 Chromium (VI) biosorption and bioaccumulation by chromate resistant bacteria; Srinath T et al.; In this study, strains that are capable of bioaccumulating Cr(VI) were isolated from treated tannery effluent of a common effluent treatment plant . The Cr(VI) concentration in this treated effluent was 0.96 mg/l, much above the statutory limit of 0.1 mg/l for discharge of industrial effluents into inland surface waters in India . In addition to the bioaccumulation, biosorption capabilities of living and dead cells were analysed . Two strains, identified as Bacillus circulans and Bacillus megaterium were able to bioaccumulate 34.5 and 32.0 mg Cr/g dry weight, respectively and brought the residual concentration of Cr(VI) to the permissible limit in 24 h when the initial concentration was 50 mg Cr(VI)/l . Our experimental design accounts for initial as well as final residual concentration of heavy metal while selecting heavy metal accumulating strains during batch studies . Biosorption of Cr(VI) was shown by B . megaterium and an another strain, B . coagulans . Living and dead cells of B . coagulans biosorbed 23.8 and 39.9 mg Cr/g dry weight, respectively, whereas, 15.7 and 30.7 mg Cr/g dry weight was biosorbed by living and dead cells of B . megaterium, respectively . Biosorption by the dead cells was higher than the living cells . This was due to prior pH conditioning (pH 2.5 with deionized water acidified with H2SO4) of the dead cells. Biomedica, 2002 Jun, 22(2), 141 - 54 {Bacillary angiomatosis}; Rodriguez G et al.; Bacillary angiomatosis is a bacterial disease which affects mainly immunosuppressed patients . It may compromise any tissue, especially the skin, presenting papules, nodules or angiomatous tumors . We studied three young men with AIDS, all of them with 1-2 papules, nodules or subcutaneous tumors suggesting telangiectatic granuloma, sarcoma and lipoma . Microscopically, they were misdiagnosed as telangiectatic granuloma, Kaposi's sarcoma and "angioma with secondary inflammation" . After reviewing the histopathology, we saw them to be composed by vessels with prominent endothelium and stroma rich in leukocytoclastic polymorphonuclears . Fibrinoid deposits were observed in the neighborhood of vessels as well as minute eosinophilic granular interstitial masses corresponding to Bartonella aggregates, criteria which answer to the diagnosis of bacillary angiomatosis with HE staining . The Warthin-Starry stain was not useful; using resin embedded tissue from paraffin-embedded material, bacterial clusters, both in semithin section stained with toluidine blue and in thin sections observed under the electron microscope, were clearly seen, confirming bacillary angiomatosis diagnosis . Patients were successfully treated with surgery and either erythromycin or doxycycline . We reviewed the entity as well as its differential diagnoses with telangiectatic granuloma, Kaposi's sarcoma, Carrion's disease, and cat-scratch disease . In conclusion, we showed the presence of bacillary angiomatosis in three patients, illustrated its typical histopathological appearance with HE staining and demonstrated the causal bacteria in thick sections and with the electron microscope . It is essential to recognize bacillary angiomatosis, as it can be cured with antibiotics. Postgrad Med J, 2002 Jun, 78(920), 327 - 9 Bacille Calmette-Guérin lymphadenitis; Goraya JS et al.; Bacille Calmette-Guerin (BCG) lymphadenitis is the most common complication of BCG vaccination . Two forms of BCG lymphadenitis can be recognised in its natural course-simple or non-suppurative lymphadenitis, which usually regresses spontaneously over a period of few weeks, and suppurative BCG lymphadenitis distinguished by the development of fluctuations in the swelling, with erythema and oedema of overlying skin . Healing in suppurative glands occurs through spontaneous perforation and sinus formation, followed by closure of the sinus by cicatrisation . Non-suppurative BCG lymphadenitis is best managed with expectant follow ups only, because medical treatment with erythromycin or antituberculous drugs do not hasten the regression or prevent development of suppuration . Suppurative BCG lymphadenitis may be treated by needle aspiration to hasten resolution and prevent spontaneous perforation and sinus formation . Surgical excision is rarely needed and is meant for cases of failed needle aspiration or for draining BCG nodes. Am J Respir Cell Mol Biol, 2002 Aug, 27(2), 244 - 9 The effect of bacillus Calmette-Guérin immunization depends on the genetic predisposition to Th2-type responsiveness; Hylkema MN et al.; The aim of this study was to investigate whether the effect of bacillus Calmette-Guerin (BCG) immunization on ovalbumin-induced allergic inflammation in a rat model depends on the genetic predisposition to react with a T helper cell (Th) 2-type cytokine response . This study was performed in an inbred Th2-predisposed "asthma prone" rat strain (brown Norway {BN}) and in an outbred nonpredisposed strain (Sprague Dawley {SD}), to differentiate between genetic and environmental factors . BCG decreased numbers of lung eosinophils and macrophages in the SD rat . This effect was not seen in the BN rat . In the BN rat, but not in the SD rat, BCG downregulated levels of total serum IgE . No significant differences were found with respect to frequencies of IFNgamma- or interleukin-4-producing cells in the lung in both rat strains . These results indicate that the degree and pathway of immunomodulatory effect of BCG in two genetically different rat strains is dependent on the genetic predisposition to develop a Th2-type response . Therefore, differences in genotype in relation to environment may result in difference in involvement of contributing pathogenic factors and thus different responsiveness to therapeutic strategies. Int J Tuberc Lung Dis, 2002 Aug, 6(8), 713 - 9 Circulation level of matrix metalloproteinase-9 is correlated with disease severity in tuberculosis patients; Hrabec E et al.; SETTING: Parenchymal lung destruction accompanied by active tuberculosis is, at least in part, caused by host as well as bacillus metalloproteinases . Mycobacterium tuberculosis has been shown to stimulate MMP-9 expression in the lung of infected organisms . DESIGN: We have used quantitative zymography and computer-assisted image analysis to measure the levels of type IV collagenases in 20 serum samples of patients with active tuberculosis and in 23 serum samples of healthy volunteers . RESULTS: Mean levels of the serum MMP-9 were over three-fold higher in tuberculous samples compared with normal serum (P < 0.0001), whereas the MMP-2 levels did not differ in these two groups . The levels of MMP-9 were significantly higher in subjects with advanced disease than in those with only limited disease changes (P < 0.05) . CONCLUSIONS: We suppose that the elevation of serum MMP-9 levels in patients with tuberculosis is affected by the augmentation of synthesis and/or secretion of this enzyme by inflammatory cells in response to M . tuberculosis infection . The observed association between the serum MMP-9 level and the extent of radiological change suggests that the quantification of the serum level of this enzyme may constitute a supplementary test in pulmonary tuberculosis diagnostics. Pediatr Res, 2002 Aug, 52(2), 148 - 54 High mobility group box chromosomal protein 1 (HMGB1) is an antibacterial factor produced by the human adenoid; Zetterstrom CK et al.; Antibacterial factors were purified from human adenoid glands by tissue extraction and consecutive steps of reversed-phase chromatography and assayed for bactericidal activity against the airway pathogen Moraxella catarrhalis and also Escherichia coli and Bacillus megaterium . One of the most active components isolated from adenoids was identified by N-terminal sequence analysis and mass spectrometry as high mobility group box chromosomal protein 1 (HMGB1) . This novel finding was further substantiated by Western blot analysis, demonstrating a protein of expected size reactive with HMGB1 antiserum . Local synthesis was confirmed by reverse-transcriptase PCR and in situ hybridization . Adenoid-derived HMGB1 and recombinant HMGB1 revealed comparable antibacterial activity at high rate . More than 95% of bacteria were eradicated within 5 min by HMGB1 in the cultures . Secretion from the adenoid gland surface was also demonstrated to contain antibacterial activity, mainly mediated by alpha-defensins, but not by HMGB1 . We conclude that HMGB1, produced and stored intracellularly in the adenoid gland, contributes to the local antibacterial barrier defense system in the upper respiratory tract. Proc Natl Acad Sci U S A, 2002 Aug 6, 99(16), 10759 - 64 Epub 2002 Jul 29. Mycobacterial heparin-binding hemagglutinin and laminin-binding protein share antigenic methyllysines that confer resistance to proteolysis; Pethe K et al.; Mycobacterium tuberculosis and Mycobacterium bovis bacillus Calmette-Guerin produce a heparin-binding hemagglutinin adhesin (HBHA) required for extrapulmonary dissemination and a laminin-binding protein (LBP) involved in cytoadherence through laminin recognition . These adhesins bear posttranslational modifications that are not present when the proteins are produced in a recombinant (r) form in Escherichia coli . Mass spectrometry analysis of HBHA revealed that the posttranslational modifications are borne by the C-terminal moiety, which comprises the heparin-binding domain made of repeated lysine-rich motifs . Amino acid sequencing showed that these modifications consist of mono- and dimethyllysines within these motifs . The methyllysine-containing repeats were recognized by mAb 4057D2 and were also detected in LBP, which is equally recognized by mAb 4057D2 . This Ab does not recognize the recombinant forms of these proteins . However, when rHBHA and rLBP were subjected to NaBH(4) and formalin treatment to induce lysine methylation, reactivity with mAb 4057D2 was recovered . Methylated rHBHA displayed enhanced resistance to proteolysis compared with rHBHA, as previously observed for native HBHA . S-adenosylmethionine-dependent HBHA methyltransferase activity was detected in the cell-wall fractions of M . bovis bacillus Calmette-Guerin and of Mycobacterium smegmatis, a species that produces LBP but naturally lacks hbhA, suggesting that the same enzyme(s) methylate(s) both LBP and HBHA . This hypothesis was confirmed by the fact that HBHA produced by recombinant M . smegmatis was also methylated . These results show that mycobacteria use enzymatic methylation of lysines to ensure greater stability of their adhesins. J Clin Microbiol, 2002 Aug, 40(8), 3053 - 6 Enterotoxigenic profiles of food-poisoning and food-borne Bacillus cereus strains; Guinebretiere MH et al.; The enterotoxigenic profiles of 51 B . cereus food-related strains were compared to those of 37 B . cereus food-poisoning strains . cytK and association of hbl-nhe-cytK enterotoxin genes were more frequent among diarrheal strains (73 and 63%) than among food-borne strains (37 and 33%) . Unlike diarrheal strains, food-borne strains showed frequent nhe and hbl gene polymorphisms and were often low toxin producers. J Clin Oncol, 2002 Aug 1, 20(15), 3242 - 8 Enhanced humoral immune response correlates with improved disease-free and overall survival in American Joint Committee on Cancer stage II melanoma patients receiving adjuvant polyvalent vaccine; DiFronzo LA et al.; PURPOSE: Although the improved overall survival (OS) of patients who receive Canvaxin (CancerVax Corp, Carlsbad, CA) polyvalent vaccine (PV) immunotherapy for metastatic melanoma has been correlated with cellular and humoral immune responses, the mechanisms of vaccine immunotherapy for early-stage melanoma are unclear . Specific immune responses to tumor-associated antigens might correlate with disease-free survival (DFS) and OS in patients receiving adjuvant PV therapy for primary melanoma . PATIENTS AND METHODS: Eighty-three patients received PV plus bacille Calmette-Guerin after wide excision of American Joint Committee on Cancer stage II melanoma . Humoral and cellular responses during the first 12 weeks of adjuvant immunotherapy were assessed by serum antibody titers to a tumor-associated 90-kd glycoprotein antigen (TA90) expressed by PV, and by delayed-type hypersensitivity (DTH) skin testing with PV (PV-DTH) . RESULTS: At a median follow-up period of 46.6 months (range, 10.7 to 93.6 months), an increased PV-DTH response seemed to be associated with improved 5-year DFS (54% v 20%) and 5-year OS (75% v 60%), but the correlations were not statistically significant . Anti-TA90 immunoglobulin (Ig) M levels > or = 1:800 were significantly correlated with improved 5-year DFS and improved 5-year OS, and multivariate analysis identified anti-TA90 IgM as an independent prognostic factor for OS and DFS . CONCLUSION: These findings suggest that an increased IgM response in patients receiving PV therapy for stage II melanoma is associated with decreased recurrence and improved survival. J Clin Oncol, 2002 Aug 1, 20(15), 3193 - 8 Phase I trial of intravesical gemcitabine in bacillus Calmette-Guérin-refractory transitional-cell carcinoma of the bladder; Dalbagni G et al.; PURPOSE: The aim of this phase I study was to determine the safety and toxicity profile of gemcitabine administered as an intravesical agent in patients with transitional-cell carcinoma (TCC) of the bladder . PATIENTS AND METHODS: Patients with superficial bladder cancer refractory to intravesical bacillus Calmette-Guerin (BCG) therapy and refusing a cystectomy were considered eligible for the trial . Gemcitabine was given in the bladder for 1 hour twice weekly in 100 mL sodium chloride for a total of six treatments . After a 1-week break, a second course of six treatments over 3 weeks was given, followed by response assessment . Four dose levels were explored: 500 mg, 1,000 mg, 1,500 mg, and 2,000 mg . RESULTS: Eighteen patients completed therapy: three at 500 mg, six at 1,000 mg, three at 1,500 mg, and six at 2,000 mg . No grade 3 or 4 toxicity was observed at 500 mg . At 1,000 mg, three patients developed hematuria and one had a skin reaction resembling grade 3 hand-foot syndrome . Three patients at 1,500 mg had no grade 3 or 4 toxicity . Of si