Annu Rev Phytopathol, 1996, 34, 275 - 97 Molecular biology of rice tungro viruses; Hull R; Rice tungro, the most important virus disease of rice in South and Southeast Asia, is caused by a complex of two viruses, rice tungro bacilliform virus (RTBV) and rice tungro spherical virus (RTSV) . RTBV is a plant pararetrovirus with bacilliform particles, the structure of which is based on T = 3 icosahedral symmetry cut across the threefold axis.The particles encapsidate a circular double-stranded DNA of 8 kbp that encodes four proteins . The current information on the properties, functions, and expression of these proteins is discussed, as is the evidence for replication by reverse transcription . Two major strains of RTBV have been recognized, one from the Indian subcontinent and the other from Southeast Asia . RTSV particles contain a single-stranded RNA genome of 12 kb that encodes a large polyprotein and possibly one or two smaller proteins . The properties and processing of the polyprotein are described and the resemblance to picornaviruses noted. Annu Rev Phytopathol, 1996, 34, 249 - 74 Biology and epidemiology of rice viruses; Hibino H; The 15 known viruses that occur in rice are rice black-streaked dwarf, rice bunchy stunt, rice dwarf, rice gall dwarf, rice giallume, rice grassy stunt, rice hoja blanca, rice necrosis mosaic, rice ragged stunt, rice stripe necrosis, rice stripe, rice transitory yellowing, rice tungro bacilliform, rice tungro spherical, and rice yellow mottle viruses . This paper describes their geographical distribution, relation to vectors, infection cycles, field dispersal, and development, and lists recorded outbreaks of the viruses . Many rice viruses have become serious problems since rice cultivation has been intensified . Double-cropping of rice using improved, photo-insensitive cultivars of short growth duration has significantly influenced the incidence of these viruses. Annu Rev Entomol, 1998, 43, 701 - 26 Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology; Gould F; This review examines potential impacts of transgenic cultivars on insect population dynamics and evolution . Experience with classically bred, insecticidal cultivars has demonstrated that a solid understanding of both the target insect's ecology and the cultivar's performance under varied field conditions will be essential for predicting area-wide effects of transgenic cultivars on pest and natural enemy dynamics . This experience has also demonstrated the evolutionary capacity of pests for adaptive response to insecticidal traits in crops . Biochemical and genetic studies of insect adaptation to the Bacillus thuringiensis (Bt) toxins expressed by currently marketed transgenic cultivars indicate a high risk for rapid adaptation if these cultivars are misused . Theoretical and practical issues involved in implementing strategies to delay pest adaptation to insecticidal cultivars are reviewed . Emphasis is placed on examining the "high dose"/refuge strategy that has become the goal of industry and regulatory authorities. Annu Rev Entomol, 1998, 43, 395 - 419 Ecology and management of hazelnut pests; AliNiazee MT; Insect and mite pests cause serious damage to the hazelnut crop worldwide . The control strategies used against these pests include application of insecticides, classical and augmentative biological control, utilization of resistant varieties, and use of Bacillus thuringiensis-based preparations . In the United States, extensive research has been directed toward elucidating ecological interactions among different pests and natural enemies and understanding the role of abiotic factors in pest population dynamics . Differences exist worldwide regarding the understanding of pests and natural enemy biologies and, to a limited extent, control practices . An integrated pest management approach based on utilizing effective sampling and monitoring techniques and a near complete reliance on biological control and "soft" pesticides, including insect growth regulators, is currently under development both in North America and Europe/Turkey . Hazelnuts are on the verge of becoming one of the first crops in the United States that could possibly be produced commercially without the use of any broad-spectrum organic insecticides. Annu Rev Entomol, 1996, 41, 451 - 72 Bacillus sphaericus toxins: molecular biology and mode of action; Charles JF et al.; Bacillus sphaericus is a spore-forming aerobic bacterium, several strains of which are pathogenic for mosquito larvae . During sporulation, the most active strains produce a crystal toxin with a high degree of larvicidal activity . The toxin is composed of two proteins of 51.4 and 41.9 kDa, which are encoded by highly conserved chromosomal genes . After B . sphaericus is ingested, these proteins are released in the larva's midgut, and, in susceptible mosquito species, bind to a specific receptor present on midgut brush-border membranes . The resulting damages to the midgut cells leads to the mosquitoes' death . During vegetative growth, some B . sphaericus strains also synthesize mosquito larvicidal proteins of 100 and 30.8 kDa (Mtx toxins), the mode of action of which is still unknown . The mechanism of acquisition of the recessive mosquito resistance to the crystal toxin varies with selection conditions. Seizure, 2004 Apr, 13(3), 191 - 5 Epilepsia partialis continua in cat scratch disease; Puligheddu M et al.; Cat scratch disease (CSD) is a world-wide, diffuse, non-epidemic infection caused by the Gram-negative bacillus Bartonella henselae . The occurrence of encephalopathy represents an infrequent and atypical complication, whose manifestations include ischemic strokes, transverse myelitis and epileptic seizures . Status epilepticus has been described as the most frequent emergency in CSD encephalopathy . In this report, we describe a case of CSD complicated by an epilepsia partialis continua (EPC) manifested as rhythmic movements of the flexor muscles of the left hand . Although CSD is a benign, self-limited disease and a complete neurological recovery usually occurs, in the present case the EPC resulted in a partial epilepsy . Magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT) and back-averaged EEG data recorded during myoclonic activity document this CSD complication. Med Vet Entomol, 2004 Mar, 18(1), 50 - 6 The naturally derived insecticide spinosad is highly toxic to Aedes and Anopheles mosquito larvae; Bond JG et al.; Spinosad is a naturally derived biorational insecticide with an environmentally favourable toxicity profile, so we investigated its potency against mosquito larvae (Diptera: Culicidae) . By laboratory bioassays of a suspension concentrate formulation of spinosad (Tracer), the 24 h lethal concentration (LC50) against Aedes aegypti (L.) third and fourth instars was estimated at 0.025 p.p.m . following logit regression . The concentration-mortality response of third- and fourth-instar Anopheles albimanus Weidemann did not conform to a logit model . The LC50 value of spinosad in Anopheles albimanus was 0.024 p.p.m . by quadratic linear regression . A field trial in southern Mexico demonstrated that spinosad 1 p.p.m . compared with the standard temephos (Abate) 1% granules 100 g/m3 water prevented Ae . aegypti breeding in plastic containers of water for 8 weeks; at 10 p.p.m . spinosad prevented breeding for > 22 weeks . In another field trial, spinosad at 5 p.p.m . and temephos both completely eliminated reproduction of Ae . aegypti for 13 weeks . In contrast, the bacterial insecticide Bacillus thuringiensis var . israelensis (Bti, Vectobac) AS) performed poorly with just 2 weeks of complete inhibition of Ae . aegypti breeding . Spinosad also effectively prevented breeding of Culex mosquitoes and chironomids in both trials to a degree similar to that of temephos . We conclude that spinosad merits evaluation as a replacement for organophosphate or Bti treatment of domestic water tanks in Mesoamerica . We also predict that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites. BJU Int, 2004 Mar, 93(4), 491 - 4 Intravesical administration of gemcitabine in superficial bladder cancer: a phase I study with pharmacodynamic evaluation; De Berardinis E et al.; OBJECTIVE: To determine, in a phase I trial, the local and systemic toxicity and pharmacodynamics of intravesical gemcitabine in patients with superficial bladder cancer . PATIENTS AND METHODS: Twelve patients with histologically confirmed carcinoma localized to the bladder wall (stage T1 or Ta) resistant to previous administration of anticancer drugs and/or of bacille Calmette-Guerin were enrolled . They initially received intravesical gemcitabine starting at 500 mg and increased in 500 mg increments to 2000 mg . Three patients were treated at each dose level . RESULTS: There was no evidence of systemic toxicity and local toxicity was minimal . A pharmacological evaluation showed that gemcitabine was undetectable in plasma and its inactive metabolite (2',2'-difluorodeoxyuridine) was present at a mean (SD) concentration of 1.39 (1.05) mumol/L Deoxycytidine kinase was present in tumour tissue samples, and its activity was 27.3 (12.6) pmol/h/mg tissue; deoxycytidine deaminase activity varied from undetectable to 616 pmol/h/mg tissue . CONCLUSION: Intravesical gemcitabine appears to be well tolerated with no systemic and minimal local toxicity even at the highest dose (2000 mg) . A phase II trial of intravesical gemcitabine at 2000 mg given weekly for six consecutive weeks is now in progress in patients with superficial bladder cancer. BJU Int, 2004 Mar, 93(4), 485 - 90 Intravesical bacillus Calmette-Guérin is superior to mitomycin C in reducing tumour recurrence in high-risk superficial bladder cancer: a meta-analysis of randomized trials; Shelley MD et al.; OBJECTIVE: To assess, in a systematic review and meta-analysis, the relative effectiveness of intravesical mitomycin C and bacillus Calmette-Guerin (BCG) for tumour recurrence, disease progression and overall survival in patients with medium- to high-risk Ta and T1 bladder cancer . METHODS: The major medical databases were searched comprehensively up to June 2003, and relevant journals hand-searched for randomized controlled trials, in any language, that compared intravesical mitomycin C with BCG in medium- to high-risk patients with Ta or T1 bladder cancer . RESULTS: Twenty-five articles were identified but only seven were considered eligible for the analysis . This represented 1901 evaluable patients in all, 820 randomized to mitomycin C and 1081 to BCG . Six trials had sufficient data for meta-analysis and included 1527 patients, 693 in the mitomycin and 834 in the BCG arm . There was no significant difference between mitomycin C and BCG for tumour recurrence in the six trials, with a weighted mean log hazard ratio, LHR, (variance) of -0.022 (0.005) . However, there was significant heterogeneity between trials (P = 0.001) . A subgroup analysis of three trials that included only high-risk Ta and T1 patients indicated no heterogeneity (P = 0.25) and a LHR for recurrence of -0.371 (0.012) . With mitomycin C used as the control in the meta-analysis, a negative ratio is in favour of BCG and, in this case, was highly significant (P < 0.001) . The seventh trial (in abstract form only) used BCG in low doses for two arms of the trial (27 mg and 13.5 mg) compared with a standard dose of mitomycin C (30 mg), and reported a significantly lower recurrence rate with BCG (27 mg) than for mitomycin C (P = 0.001) . Only two trials included sufficient data to analyse disease progression and survival, representing 681 patients (338 randomized to BCG and 343 to mitomycin C) . There was no significant difference between mitomycin C and BCG for disease progression, with a LHR of 0.044 (0.04) (P = 0.16), or survival, at -0.112 (0.03) (P = 0.50) . Adverse events were slightly more frequent with BCG . Local toxicity (dysuria, cystitis, frequency and haematuria) were associated with both mitomycin C (30%) and BCG (44%) . Systemic toxicity, e.g . chills, fever and malaise, occurred with both agents (12% and 19%, respectively) although skin rash was more common with mitomycin C . CONCLUSION: Tumour recurrence was significantly lower with intravesical BCG than with mitomycin C only in those patients at high risk of tumour recurrence . However, there was no difference in disease progression or survival, and the decision to use either agent might be based on adverse events and cost. Appl Environ Microbiol, 2004 Mar, 70(3), 1514 - 21 Construction of a functional S-layer fusion protein comprising an immunoglobulin G-binding domain for development of specific adsorbents for extracorporeal blood purification; Vollenkle C et al.; The chimeric gene encoding a C-terminally-truncated form of the S-layer protein SbpA from Bacillus sphaericus CCM 2177 and two copies of the Fc-binding Z-domain was constructed, cloned, and heterologously expressed in Escherichia coli HMS174(DE3) . The Z-domain is a synthetic analogue of the B-domain of protein A, capable of binding the Fc part of immunoglobulin G (IgG) . The S-layer fusion protein rSbpA(31-1068)/ZZ retained the specific properties of the S-layer protein moiety to self-assemble in suspension and to recrystallize on supports precoated with secondary cell wall polymer (SCWP), which is the natural anchoring molecule for the S-layer protein in the bacterial cell wall . Due to the construction principle of the S-layer fusion protein, the ZZ-domains remained exposed on the outermost surface of the protein lattice . The binding capacity of the native or cross-linked monolayer for human IgG was determined by surface plasmon resonance measurements . For batch adsorption experiments, 3-microm-diameter, biocompatible cellulose-based, SCWP-coated microbeads were used for recrystallization of the S-layer fusion protein . In the case of the native monolayer, the binding capacity for human IgG was 5.1 ng/mm(2), whereas after cross-linking with dimethyl pimelimidate, 4.4 ng of IgG/mm(2) was bound . This corresponded to 78 and 65% of the theoretical saturation capacity of a planar surface for IgGs aligned in the upright position, respectively . Compared to commercial particles used as immunoadsorbents to remove autoantibodies from sera of patients suffering from an autoimmune disease, the IgG binding capacity of the S-layer fusion protein-coated microbeads was at least 20 times higher . For that reason, this novel type of microbeads should find application in the microsphere-based detoxification system. Appl Environ Microbiol, 2004 Mar, 70(3), 1378 - 84 Interaction of Bacillus thuringiensis toxins with larval midgut binding sites of Helicoverpa armigera (Lepidoptera: Noctuidae); Estela A et al.; In 1996, Bt-cotton (cotton expressing a Bacillus thuringiensis toxin gene) expressing the Cry1Ac protein was commercially introduced to control cotton pests . A threat to this first generation of transgenic cotton is the evolution of resistance by the insects . Second-generation Bt-cotton has been developed with either new B . thuringiensis genes or with a combination of cry genes . However, one requirement for the "stacked" gene strategy to work is that the stacked toxins bind to different binding sites . In the present study, the binding of (125)I-labeled Cry1Ab protein ((125)I-Cry1Ab) and (125)I-Cry1Ac to brush border membrane vesicles (BBMV) of Helicoverpa armigera was analyzed in competition experiments with 11 nonlabeled Cry proteins . The results indicate that Cry1Aa, Cry1Ab, and Cry1Ac competed for common binding sites . No other Cry proteins tested competed for either (125)I-Cry1Ab or (125)I-Cry1Ac binding, except Cry1Ja, which competed only at the highest concentrations used . Furthermore, BBMV from four H . armigera populations were also tested with (125)I-Cry1Ac and Cry1Ab to check the influence of the insect population on the binding results . Finally, the inhibitory effect of selected sugars and lectins was also determined . (125)I-Cry1Ac binding was strongly inhibited by N-acetylgalactosamine, sialic acid, and concanavalin A and moderately inhibited by soybean agglutinin . In contrast, (125)I-Cry1Ab binding was only significantly inhibited by concanavalin A . These results show that Cry1Ac and Cry1Ab use different epitopes for binding to BBMV. Clin Exp Allergy, 2004 Mar, 34(3), 398 - 405 Bacillus Calmette-Guérin-induced interleukin-12 did not additionally improve clinical and immunologic parameters in asthmatic children treated with sublingual immunotherapy; Arikan C et al.; OBJECTIVE: To evaluate the effect of bacillus Calmette-Guerin (BCG) as an adjuvant to specific sublingual immunotherapy (SLIT) on the cytokine profile of peripheral blood mononuclear cells (PBMCs) and clinical outcome . METHODS: Thirty-two children with asthma and rhinitis allergic to house dust mite (HDM) with negative purified protein derivative (PPD) skin test response were enrolled . After a run-in period of 8 weeks, patients were randomized to receive either SLIT only (n=16) or one dose of BCG immunization before initiation of SLIT (n=16) with a standardized Dermatophagoides pteronyssinus (D . pteronyssinus)+D . farinea 50/50 extract . PPD-negative asthmatics (n=5) allergic to HDM receiving inhaled therapy only were included for comparison of cytokine levels in PBMC cultures . Efficacy was assessed both at the end of run-in and 6 months of treatment periods with criteria including symptom, medication and quality-of-life (QoL) scores, IgE levels, lung function, provocation concentration (PC20), eosinophil count and skin prick tests . IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-gamma levels were determined in antigen specifically and polyclonally stimulated PBMC cultures . RESULTS: Both treatment groups showed significant improvement at the end of 6 months for asthma and rhinitis scores and QoL, number of asthma attacks, amount of beta2-agonists, inhaled and intranasal steroids, blood eosinophil counts and PC20 . Interestingly, phytohaemagglutinin (PHA)-stimulated IL-12 and D . pteronyssinus-stimulated IFN-gamma in PBMC were significantly higher in the treatment groups than controls . In addition, IL-12 levels in response to D . pteronyssinus and PHA stimulation were significantly higher in the SLIT+BCG group than the SLIT alone group and controls . CONCLUSION: The present study demonstrates that successful SLIT is parallel to increased IFN-gamma production by PBMC . Although simultaneous BCG vaccination enhanced IL-12 production, it did not additionally improve the clinical outcome. Biochemistry, 2004 Mar 16, 43(10), 2784 - 91 The crystal structure of glutamyl endopeptidase from Bacillus intermedius reveals a structural link between zymogen activation and charge compensation; Meijers R et al.; Extracellular glutamyl endopeptidase from Bacillus intermedius (BIEP) is a chymotrypsin-like serine protease which cleaves the peptide bond on the carboxyl side of glutamic acid . Its three-dimensional structure was determined for C222(1) and C2 crystal forms of BIEP to 1.5 and 1.75 A resolution, respectively . The topology of BIEP diverges from the most common chymotrypsin architecture, because one of the domains consists of a beta-sandwich consisting of two antiparallel beta-sheets and two helices . In the C2 crystals, a 2-methyl-2,4-pentanediol (MPD) molecule was found in the substrate binding site, mimicking a glutamic acid . This enabled the identification of the residues involved in the substrate recognition . The presence of the MPD molecule causes a change in the active site; the interaction between two catalytic residues (His47 and Ser171) is disrupted . The N-terminal end of the enzyme is involved in the formation of the substrate binding pocket . This indicates a direct relation between zymogen activation and substrate charge compensation. Biochim Biophys Acta, 2004 Mar 9, 1661(2), 154 - 65 Highly robust lipid membranes on crystalline S-layer supports investigated by electrochemical impedance spectroscopy; Gufler PC et al.; In the present work, S-layer supported lipid membranes formed by a modified Langmuir-Blodgett technique were investigated by electrochemical impedance spectroscopy (EIS) . Basically two intermediate hydrophilic supports for phospholipid- (DPhyPC) and bipolar tetraetherlipid- (MPL from Thermoplasma acidophilum) membranes have been applied: first, the S-layer protein SbpA isolated from Bacillus sphaericus CCM 2177 recrystallized onto a gold electrode; and second, as a reference support, an S-layer ultrafiltration membrane (SUM), which consists of a microfiltration membrane (MFM) with deposited S-layer carrying cell wall fragments . The electrochemical properties and the stability of DPhyPC and MPL membranes were found to depend on the used support . The specific capacitances were 0.53 and 0.69 microF/cm(2) for DPhyPC bilayers and 0.75 and 0.77 microF/cm(2) for MPL monolayers resting on SbpA and SUM, respectively . Membrane resistances of up to 80 mega Ohm cm(2) were observed for DPhyPC bilayers on SbpA . In addition, membranes supported by SbpA exhibited a remarkable long-term robustness of up to 2 days . The membrane functionality could be demonstrated by reconstitution of membrane-active peptides such as valinomycin and alamethicin . The present results recommend S-layer-supported lipid membranes as promising structures for membrane protein-based biosensor technology. J Microbiol Methods, 2004 Apr, 57(1), 1 - 7 Evaluation of the MicroFoss system for the detection of Listeria species in environmental samples; Odumeru JA et al.; The MicroFoss system was evaluated for its ability to detect Listeria species in environmental samples . The sensitivity and specificity of the MicroFoss were determined in relation to a standard culture method for Listeria detection . The sensitivities of both the MicroFoss and standard culture methods were similar (88.4%-MicroFoss, 90.7%-Culture) based on the total number of positive results obtained by both methods . The MicroFoss system detected Listeria spp . in 12 samples, which were not detected by culture, and the culture method detected Listeria spp . in 15 samples, which were not detected by the MicroFoss method . This was likely due to uneven distribution of low levels of Listeria organisms in the split sponge samples used to assess the performance of these test methods . The specificity value determined for the MicroFoss system was 92.7% . The majority of microbes causing false positive results in the MicroFoss system were Bacillus species, which were readily distinguishable from Listeria species by a simple Gram stain and morphological features . Listeria monocytogenes (89.4%-MicroFoss, 88.0%-Culture) and Listeria innocua (8.8%-MicroFoss, 7.7%-Culture) were the most common isolates of Listeria detected by the two test methods, with L . monocytogenes being the most predominant isolate detected . The highly comparable results and rapid nature of the MicroFoss system demonstrate its effectiveness as a detection system for species of Listeria in environmental samples . The fact that the sensitivity of the MicroFoss system was similar to that of the culture method and the Listeria results were obtained within 48 h of testing, support the use of the MicroFoss as an alternative rapid method for screening large numbers of environmental samples for Listeria spp. Appl Spectrosc, 2004 Feb, 58(2), 203 - 11 Identification of sporulated and vegetative bacteria using statistical analysis of fourier transform mid-infrared transmission data; Foster NS et al.; A combined mid-infrared spectroscopic/statistical modeling approach for the discrimination and identification, at the strain level, of both sporulated and vegetative bacterial samples is presented . Transmission mode spectra of bacteria dried on ZnSe windows were collected using a Fourier transform mid-infrared (FT-IR) spectrometer . Five Bacillus bacterial strains (B . atrophaeus 49337, B . globigii Dugway, B . thuringiensis spp . kurstaki 35866, B . subtilis 49760, and B . subtilis 6051) were used to construct a reference spectral library and to parameterize a four-step statistical model for the systematic identification of bacteria . The statistical methods used in this initial feasibility study included principal component analysis (PCA), classification and regression trees (CART), and Mahalanobis distance calculations . Internal cross-validation studies successfully classified 100% of the samples into their correct physiological state (sporulated or vegetative) and identified 67% of the samples correctly as to their bacterial strain . Analysis of thirteen blind samples, which included reference and other bacteria, nonbiological materials, and mixtures of both nonbiological and bacterial samples, yielded comparable accuracy . The primary advantage of this approach is the accurate identification of unknown bacteria, including spores, in a matter of minutes. Biochemistry (Mosc), 2004 Feb, 69(2), 181 - 7 Reconstruction of Bacillus thuringiensis ssp . israelensis Cry11A endotoxin from fragments corresponding to its N- and C-moieties restores its original biological activity; Revina LP et al.; Subtilisin hydrolyzes Cry11A endotoxin (of 70 kD) produced by Bacillus thuringiensis ssp . israelensis to fragments of 33- and 36-kD, which correspond to N- and C-terminal halves of the endotoxin molecule . Thermitase (a serine protease from Thermoactinomyces vulgaris) and insect gut proteases from Diptera and Lepidoptera exhibit the same hydrolytic effect on Cry11A . Hydrolyzates maintain high toxicity with respect to larvae of Aedes aegypti, Anopheles stephensi, and Culex pipiens . The 33- and 36-kD Cry11A endotoxin components purified by ion-exchange chromatography from the subtilisin hydrolyzate were inactive; however, equimolar mixture of these proteins exhibited almost the same activity as the initial hydrolyzate. Biotechnol Lett, 2004 Jan, 26(2), 143 - 5 Bacillus thuringiensis var . israelensis production involving re-use of the supernatant; Luna CL et al.; The supernatant arising after biomass separation of Bacillus thuringiensis var . israelensis by flocculation/sedimentation was re-used after being supplemented with 25, 50 and 75% (w/v) of the original culture medium, based on corn steep liquor, glucose and mineral salts . Supplementation at 75% gave a spore concentration (1 x 10(10) c.f.u . ml(-1)) five times greater than that obtained with the other supplements. Biotechnol Lett, 2004 Jan, 26(2), 115 - 9 Purification and characterization of a new peptide antibiotic produced by a thermotolerant Bacillus licheniformis strain; Mendo S et al.; A Bacillus licheniformis strain, 189, isolated from a hot spring environment in the Azores, Portugal, strongly inhibited growth of Gram-positive bacteria . It produced a peptide antibiotic at 50 degrees C . The antibiotic was purified and biochemically characterized . It was highly resistant to several proteolytic enzymes . Additionally, it retained its antimicrobial activity after incubation at pH values between 3.5 and 8; it was thermostable, retaining about 85% and 20% of its activity after 6 h at 50 degrees C and 100 degrees C, respectively . Its molecular mass determined by mass spectrometry was 3249.7 Da. J Econ Entomol, 2004 Feb, 97(1), 51 - 8 Toxicity of VectoLex (Bacillus sphaericus) products to selected Australian mosquito and nontarget species; Brown ID et al.; Laboratory and field bioassay studies were conducted in southeast Queensland, Australia, on the efficacy of VectoLex Control Granule (CG; active ingredient {AI}:50 Bacillus sphaericus {B.s.} International Toxic Units {ITU}/mg) and VectoLex Water Dispersible Granule (WDG) (AI: 650 B.s . ITU/mg) formulations against third-instar larvae of Culex annulirostris Skuse, Culex quinquefasciatus Say, Culex sitiens Wiedemann, Ochlerotatus rigilax (Skuse), Ochlerotatus . notoscriptus (Skuse), and Aedes aegypti (L.) . The Gompertz model for log-linear mortality data were used to determine laboratory 48-h LC95 values . As with foreign evaluations, the B.s . formulations were most effective against Culex spp., with the WDG 10-100 times more effective than the CG on an ITU/mosquito basis . Consequently, Cx . annulirostris and Cx . quinquefasciatus were selected as target species for small-plot field evaluation of WDG efficacy over time . Weekly cohorts of caged third-instar Cx . annulirostris were exposed to replicated low (250 g/ha), medium (500 g/ha), and high (1,000 g/ha) dosages of WDG . Concurrent assessment of Cx . quinquefasciatus mortality outside the cages was also conducted . In water with high organic content, the low rate produced > 99% Cx . annulirostris mortality at 48 h, decreasing to 79% at week 3 and no control at week 4 . The medium and high rates resulted in 100% Cx . annulirostris mortality for 2 wk posttreatment, decreasing to 95% at week 3, and no control at week 4 . The WDG was equally effective against Cx . quinquefasciatus . Treatment did not affect water quality or nontarget shrimp and fish species survival. Ying Yong Sheng Tai Xue Bao, 2003 Nov, 14(11), 2062 - 6 {Advances in safety studies of soil Bt toxin proteins released from transgenic Bt crops}; Bai Y et al.; Commercialized transgenic Bt (Bacillus thuringiensis) crops are permitted for field growth in a large scale, which leads to significant issues of ecological risk assessment in soil ecosystem . In this paper, some general safety problems involving in the soil Bt active toxins released from insect-resistant transgenic Bt crops in the forms of plant residues, root exudates and pollens were reviewed, including their adsorption by soil active-particles, their insecticidal activity, persistence, and biodegradation by soil microbes, and their effects on soil organisms. J Biol Chem, 2004 May 7, 279(19), 20490 - 500 Epub 2004 Mar 02. Cross-linking phosphatidylinositol-specific phospholipase C traps two activating phosphatidylcholine molecules on the enzyme; Zhang X et al.; Bacillus thuringiensis phosphatidylinositol-specific phospholipase C (PI-PLC), a bacterial model for the catalytic domain of mammalian PI-PLC enzymes, was cross-linked by 1-ethyl-3-{3-dimethylaminopropyl}carbodiimide hydrochloride to probe for the aggregation and/or conformational changes of PI-PLC when bound to activating phosphatidylcholine (PC) interfaces . Dimers and higher order multimers (up to 31% of the total protein when cross-linked at pH 7) were observed when the enzyme was cross-linked in the presence of PC vesicles . Aggregates were also detected with PI-PLC bound to diheptanoyl-PC (diC(7)PC) micelles, although the fraction of cross-linked multimers (19% at pH 7) was lower than when the enzyme was cross-linked in the presence of vesicles . PI-PLC cross-linked in the presence of a diC(7)PC interface exhibited an enhanced specific activity for PI cleavage . The extent of this cross-linking-enhanced activation was reduced in PI-PLC mutants lacking either tryptophan in the rim (W47A and W242A) of this (betaalpha)(8)-barrel protein . The higher activity of the native protein cross-linked in the presence of diC(7)PC correlated with an increased affinity of the protein for two diC(7)PC molecules as detected by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry . In contrast to wild type protein, W47A and W242A had only a single diC(7)PC tightly associated when cross-linked in the presence of that activator molecule . These results indicate that (i) each rim tryptophan residue is involved in binding a PC molecule at interfaces, (ii) the affinity of the enzyme for an activating PC molecule is enhanced when the protein is bound to a surface, and (iii) this conformation of the enzyme with at least two PC bound that is stabilized by chemical cross-linking interacts more effectively with activating interfaces, leading to higher observed specific activities for the phosphotransferase reaction. Int J Food Microbiol, 2004 Mar 1, 91(2), 209 - 13 Effect of high pressure gaseous carbon dioxide on the germination of bacterial spores; Furukawa S et al.; Effect of high pressure gaseous carbon dioxide treatment (HGCT) at 6.5 MPa, 35 degrees C on the germination of bacterial spores was investigated . Germination of bacterial spores was estimated by the decrease of heat tolerance . Approximately, 40% of Bacillus coagulans and 70% of Bacillus licheniformis were germinated by HGCT for 120 min at 35 degrees C, respectively . Germination was confirmed by phase contrast microscopy . The effect of hydrostatic pressure treatment (HPT) at 6.5 MPa, 35 degrees C on the germination of B . coagulans and B . licheniformis spores were also investigated . Spores did not germinate by HPT alone at 6.5 MPa for 120 min. Transfusion, 2004 Mar, 44(3), 337 - 42 The use of a bacteria detection system to evaluate bacterial contamination in PLT concentrates; Rock G et al.; BACKGROUND: Random-donor PLTs (RDPs) are functional at 7 days . Nevertheless, since the mid-1980s, concern for bacterial contamination has caused the storage period to be reduced to 5 days . The ability of a bacteria detection system (BDS, Pall) to determine bacterial contamination and permit extension of the PLT shelf life to 7 days was assessed . STUDY DESIGN AND METHODS: Blood was collected into CP2D and leukoreduced RDPs were prepared . Upon arrival at the hospital, a 2- to 3-mL aliquot was removed from each RDP and introduced into the Pall BDS pouch with a sterile docking device . The pouch was incubated at 37 degrees C for 24 hours and then the oxygen content was measured to determine bacterial contamination . Additionally, the RDPs were pooled and an aliquot was removed for culture with standard manual techniques . CCIs were calculated 1 hour after infusion . RESULTS: A total of 12,062 individual RDPs were tested . The Pall BDS detected bacteria in 5 units . All of these were positive on repeat sampling . Propionibacterium acnes, coagulase-negative Staphylococcus, and Bacillus species were confirmed by manual technique in 3 units, one could not be identified, and one was negative . Aliquots from PLT pools were positive in 80 of 2201 pools when tested by manual methods . Of these, 79 were false-positives and 1 unit contained coagulase-negative Staphylococcus . The Pall BDS was easy to use and required less than 5 minutes for all manipulations . After 7 days of storage, the PLTs gave an average CCI of 16 x 10(11)+/- 3.39 x 10(11) 1 hour after transfusion (n = 9) . CONCLUSIONS: The Pall BDS permits evaluation of RDPs for bacterial contamination . Culture-negative PLTs were successfully transfused in our institution up to and including 7 days after storage with good CCIs. J Agric Food Chem, 2004 Mar 10, 52(5), 1390 - 7 Lower fumonisin mycotoxin levels in the grain of Bt corn grown in the United States in 2000-2002; Hammond BG et al.; Fumonisins were monitored in corn grain collected from Bt hybrids grown in 107 locations across the United States in 2000-2002 . Bt corn hybrids contain the Cry1Ab protein from Bacillus thuringiensis that controls European corn borers and other stalk-boring pests . Fumonisin levels were frequently lower in grain from Bt hybrids grown in field trials under conditions of natural (FACT trials) or manual insect infestation (university trials) . Over three years of FACT trials, there were 126/210 comparisons when fumonisin levels in grain from control hybrids were >2 ppm, exceeding U.S . FDA guidance levels of 2 ppm for human food . Grain from Bt hybrids was at or below 2 ppm of fumonisins for 58 of the 126 comparisons . The use of Bt hybrids can increase the percentage of corn grain that would be suitable for use in food and feed. J Econ Entomol, 2003 Apr, 96(2), 396 - 400 Laboratory and field evaluation of tebufenozide, diflubenzuron, and Bacillus thurengiensis var . kurstaki for suppression of Douglas-fir tussock moth (Orgyia pseudotsugata (McDunnough)) in Idaho: a case study; Cook SP; The insect growth regulator tebufenozide (MIMIC 2LV) was tested to examine its impact on the Douglas-fir tussock moth, Orgyia pseudotsugata (McDunnough) (Lepidoptera: Lymantriidae) . Laboratory tests gave an estimated concentration of 1.26 ppm of the compound to achieve 50% population mortality (LC50) for second-instar O . pseudotsugata . At the highest concentration tested, tebufenozide resulted in significant larval mortality within 7 d with an estimated time to 50% population mortality (LT50) of 6.3 d . A field comparison oftebufenozide with diflubenzuron (Dimilin 4L) and Bacillus thurengiensis var . kurstaki (Btk, FORAY 48B) was also conducted . There was no significant difference in larval mortality within field plots that were treated with diflubenzuron (42.4%) or Btk (44.8%) . Larval mortality in the tebufenozide-treated plots (56.8%) was also similar to the mortality in the diflubenzuron and Btk treatments . All three treatments resulted in more larval mortality than that measured in untreated controlplots (11.2%) . Both tebufenozide and diflubenzuron treatments resulted in significantly more mortality (55.0% and 40.0%, respectively) to larvae-fed treated foliage 3 wk after application than was measured for larvae-fed foliage from untreated trees (11.0%) . There was no significant difference among the treatments in the percentage of host trees in the overstory that sustained >25% defoliation, and all three treatments resulted in less defoliation than was measured in the control plots . There was no significant difference among the treatments in the percentage of host trees in the understory that sustained >25% defoliation. J Econ Entomol, 2003 Apr, 96(2), 388 - 95 Design and evaluation of an aerial spray trial with true replicates to test the efficacy of Bacillus thuringiensis insecticide in a boreal forest; Cadogan BL et al.; A field trial using true replicates was conducted successfully in a boreal forest in 1996 to evaluate the efficacy of two aerially applied Bacillus thuringiensis formulations, ABG 6429 and ABG 6430 . A complete randomized design with four replicates per treatment was chosen . Twelve to 15 balsam fir (Abies balsamea {L.} Mill.) per plot were randomly selected as sample trees . Interplot buffer zones, > or = 200 m wide, adequately prevented cross contamination from sprays that were atomized with four rotary atomizers (volume median diameters ranging from 64.6 to 139.4 microm) and released approximately 30 m above the ground . The B . thuringiensis formulations were not significantly different (P > 0.05) from each other in reducing spruce budworm (Choristoneura fumiferana {Clem.}) populations and protecting balsam trees from defoliation but both formulations were significantly more efficacious than the controls . The results suggest that true replicates are a feasible alternative to pseudoreplication in experimental forest aerial applications. J Econ Entomol, 2003 Apr, 96(2), 292 - 9 Preparation of spray-dried wettable powder formulations of Bacillus thuringiensis-based biopesticides; Teera-Arunsiri A et al.; Bacillus thuringiensis is the most widely used biopesticide among many methods available to control insects . To make a saleable product, B . thuringiensis must be substantially concentrated by removal of water and formulated to improve longevity, efficacy, and ease of transport of the product . B . thuringiensis subsp . aizawai culture broth as an active ingredient was mixed with various adjuvants and then spray dried . The optimum conditions for spray drying were found to be an outlet temperature of 60-85 degrees C and an inlet temperature of 120-180 degrees C . Various adjuvants had different effects on physical and biological properties of the dried product . Gelatinized tapioca starch and milk powder improved suspensibility but adversely affected wettability of the dried formulated product . Vegetable oil and Tween 20 enhanced wettability but resulted in poor suspensibility . Silica fume was used to enhance flowability because it reduced clumping and caking of the powder resulting from the addition of vegetable oil . Formulation containing 10% wt:wt B . thuringiensis, 10% wt:wt gelatinized tapioca starch, 10% wt:wt sucrose, 38% wt:wt tapioca starch, 20% wt:wt milk powder, 10% wt:wt silica fume, 2% wt:wt polyvinyl alcohol, 5% vol:vol Tween 20, 1% vol:vol refined rice bran oil, and 1% vol:vol antifoam solution was found to be optimum in terms of the physical and biological properties of the dried product . This formulation had 55% suspensibility, 24 s for wetting time, and 5.69 x 10(4) CFU/ml of LC50 value against Spodoptera exigua larvae. J Econ Entomol, 2003 Apr, 96(2), 280 - 6 Lethal and sublethal effects of single and double applications of Bacillus thuringiensis variety kurstaki on spruce budworm (Lepidoptera: Tortricidae) larvae; Moreau G et al.; We conducted laboratory experiments to examine the effects of single versus double exposures of spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae) female larvae to various concentrations of a Bacillus thuringiensis variety kurstaki (Btk) commercial formulation (Foray 48B) . Our main objective was to document the vulnerability to Btk and the sublethal responses of fifth-instar larvae that survived from a first ingestion of Btk during their fourth stadium and to compare them with insects treated either during their fifth or fourth stadium only . As reported in the literature, fifth-instar larvae were more vulnerable than fourth-instar larvae, but only at low and medium concentrations . Fifth-instar larvae that had survived Btk ingestion during their fourth stadium were more vulnerable to a high concentration of Btk and had a shorter feeding inhibition period than those that had not been exposed during their fourth stadium . Compared with a single treatment at the fourth stadium, a double exposure to Btk further reduced the population by 20-30%, depending on the concentration applied . The second treatment also induced another feeding inhibition period and increased larval development time by 14% . The impact of the different treatments on pupal weight depended on whether treated insects exhibited supernumerary instars . In the absence of developmental polymorphism, a higher concentration, a late, or a double exposure to Btk significantly reduced pupal weight. Acta Crystallogr D Biol Crystallogr, 2004 Mar, 60(Pt 3), 586 - 7 Epub 2004 Feb 25. Crystallization and preliminary X-ray study of gamma-type cyclodextrin glycosyltransferase from Bacillus clarkii; Akita M et al.; A gamma-cyclodextrin glycosyltransferase (EC 2.4.1.19) from Bacillus clarkii was crystallized using the hanging-drop vapour-diffusion method at 293 K . X-ray diffraction data were collected to 2.2 A . The crystal belongs to space group R3, with unit-cell parameters a = b = 211.6, c = 52.7 A . The asymmetric unit contains one protein molecule, with a corresponding V(M) of 3.03 A(3) Da(-1) and a solvent content of 59.4% . Molecular replacement was successfully carried out using a homology model based on the three-dimensional structure of the CGTase from Thermonanaerobacterium thermosulfurigenes EM1 as a search model. Acta Crystallogr D Biol Crystallogr, 2004 Mar, 60(Pt 3), 564 - 6 Epub 2004 Feb 25. Crystallization and preliminary X-ray crystallographic analysis of the C-terminal domain of ParC protein from Bacillus stearothermophilus; Hsieh TJ et al.; Type IIA topoisomerases are multidomain enzymes composed of four major domains: the ATPase domain, the TOPRIM domain, the DNA-cleavage/religation domain and the C-terminal domain (CTD) . Although crystal structures of the first three domains are available, the three-dimensional structure of the less-conserved CTD has yet to be determined . In order to provide a three-dimensional structure of this structurally uncharacterized region, the 36 kDa CTD of ParC protein, the DNA-cleavage/religation subunit of topoisomerase IV, from Bacillus stearothermophilus has been cloned, purified and crystallized . The crystals belonged to the trigonal space group P3(1) (or P3(2)), with unit-cell parameters a = b = 83.5, c = 45.1 A . The asymmetric unit contains one molecule and the solvent content is 51.2% . A 98.9% complete native data set has been collected from a frozen crystal to 2.0 A resolution with an overall R(merge) of 6.5%. Am J Trop Med Hyg, 2004 Feb, 70(2), 197 - 200 Parallel assessment of 24 monthly doses of rifampin, ofloxacin, and minocycline versus two years of World Health Organization multi-drug therapy for multi-bacillary leprosy; Villahermosa LG et al.; Monthly doses of rifampin, ofloxacin, and minocycline (ROM) are expected to be effective treatment for multi-bacillary leprosy . Patients with MB leprosy received ROM (n = 10) or World Health Organization multi-drug therapy (MDT) (n = 11) . Treatment with ROM was given as 24 consecutive monthly observed doses of rifampin (600 mg), ofloxacin (400 mg), and minocycline (100 mg) . Treatment with MDT was given as 24 consecutive monthly observed doses of rifampin (600 mg) and clofazimine (300 mg), and unobserved daily dapsone (100 mg) and clofazimine (50 mg) . Twenty patients completed the 24-month regimens with > 99% compliance . Treatments with ROM and MDT were safe, tolerable, and caused similar improvements in lesions, bacterial indices, and histology . All MDT recipients developed clofazimine-induced pigmentation . Six ROM and nine MDT recipients assessed at five or more years after completion of treatment had no evidence of relapse . Twenty-four months of treatment with ROM is a safe, well-tolerated, and convenient regimen that may provide an alternate therapy to MDT for MB leprosy . Larger trials with sufficient follow-up would better define the role of ROM. J Med Entomol, 2004 Jan, 41(1), 58 - 64 Inheritance and mechanism of resistance to Bacillus sphaericus in Culex quinquefasciatus (Diptera: Culicidae) from China and Brazil; Oliveira CM et al.; Investigations on the inheritance and mechanism of resistance to Bacillus sphaericus Neide in Culex quinquefasciatus Say colonies, selected with strains C3-41 (RLCq1/C3-41) and 2362 (CqRL1/2362), were performed in China and Brazil, respectively . The progeny of reciprocal F1 crosses (susceptible female x resistant male and vice versa) from both resistant colonies responded alike in bioassays, indicating recessive inheritance . Data on larvae susceptibility from the backcross offspring between F1 and their respective susceptible and resistant parental colonies are consistent with a monofactorial and autosomal mode of inheritance . In vitro binding assays between 125I binary (Bin2) toxin and the brush border membrane fractions (BBMF) from CqRL1/2362 and RLCq1/C3-41 larvae showed that resistance, in both colonies, is caused by a failure in the binding step of the B . sphaericus Bin2 toxin to its specific midgut receptor . The specific and saturable binding of Bin2 toxin to BBMF from F1 larvae (CqRL1/2362 X susceptible counterpart) confirms the recessive inheritance of the resistance gene . Further studies are needed to advance understanding of B . sphaericus resistance. FEMS Microbiol Lett, 2004 Feb 16, 231(2), 191 - 6 The Erp protein is anchored at the surface by a carboxy-terminal hydrophobic domain and is important for cell-wall structure in Mycobacterium smegmatis; Kocincova D et al.; Erp (Exported Repetitive Protein), also known as P36, Pirg and Rv3810, is a member of a mycobacteria-specific family of extracellular proteins . In pathogenic species, the erp gene has been described as a virulence factor . The Erp proteins comprise three domains . The N- and C-terminal domains are similar in all mycobacterial species, while the central domain consists of a repeated module that differs considerably between species . Here we show that the Erp protein is loosely attached to the surface and that the carboxy-terminal domain, which displays hydrophobic features, anchors Erp at the surface of the bacillus . The hydrophobic region is not necessary for the complementation of the altered colony morphology of a Mycobacterium smegmatis erp- mutant but proved to be necessary to achieve resistance to detergent at wild-type levels. Clin Exp Allergy, 2004 Feb, 34(2), 207 - 12 Effect of repeated intradermal injections of heat-inactivated Mycobacterium bovis bacillus Calmette-Guérin in adult asthma; Shirtcliffe PM et al.; BACKGROUND: There are a number of reports in the Chinese medical literature from the last 30 years regarding the efficacy of repeated doses of heat-inactivated bacillus Calmette-Guerin (BCG) in established asthma . There is also epidemiological and experimental evidence that exposure to mycobacteria has the potential to suppress the development of asthma/atopy . METHODS: Thirty-one Mantoux-negative adults with stable moderately severe asthma who were skin prick test positive to house dust mite were randomized to receive one injection (0.1 mL) a week for 4 weeks of heat-inactivated BCG or normal saline . Markers of asthma severity (including peak flow, forced expiratory volume in 1 s, major and minor exacerbations, symptom scores and beta-agonist use), blood eosinophil and IgE levels were monitored for 3 months . RESULTS: There were no statistically significant differences between the treatment group and placebo for any of the outcome variables . The recruitment to the trial was halted early and the number of injections reduced in a number of patients due to excessive local reactions to BCG . CONCLUSIONS: In addition to the lack of efficacy of repeated heat-inactivated BCG injections, the occurrence of severe local reactions will limit the therapeutic application of this approach in asthma. Transpl Infect Dis, 2003 Dec, 5(4), 195 - 8 Oerskovia xanthineolytica endocarditis in a renal transplant patient: case report and review of the literature; Urbina BY et al.; Oerskovia species were, until recently, only rarely associated with human disease . This gram-positive bacillus can be easily misidentified as a diphtheroid, a common contaminant in blood cultures . There have been 17 reports of invasive Oerskovia infection in immunocompromised hosts . We report a case of Oerskovia xanthineolytica endocarditis in a renal transplant patient and review the microbiologic and clinical characteristics of this potential pathogen. Prescrire Int, 2003 Dec, 12(68), 226 - 9 Intradermal BCG: partial protection against tuberculosis in children: unproven efficacy of multipuncture administration; The immunological aspects of latency in tuberculosis; Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA . jchan@aecom.yu.edu A unique feature of Mycobacterium tuberculosis is its ability to exist in the granuloma of an asymptomatic host in a latent state that can subsequently reactivate to cause active disease . The latent state of infection poses a major obstacle to eradicating tuberculosis . In latent tuberculosis, the host immune response is capable of controlling the infection and yet falls short of eradicating the pathogen . That the host immune response contributes to the maintenance of latent tuberculous infection is supported by the observation that certain immunodeficient states, including those associated with the human immunodeficiency virus and tumor necrosis factor neutralization therapy, are associated with increased risks for developing reactivation disease . Latent tuberculosis is the product of a complex set of interactions between M . tuberculosis and the host immune response . The molecular basis for the persistence phenotype of M . Tuberculosis and the pertinent host immune mechanisms that contribute to the maintenance of tuberculous latency are just beginning to be understood . This review discusses the interactions between M . tuberculosis and the macrophage, the primary host cell that the tubercle bacillus parasitizes. J Int Assoc Physicians AIDS Care (Chic Ill), 2003 Oct-Dec, 2(4), 133 - 9 Clinical vignette in antiretroviral therapy: jaundice; Zell SC; HIV caregivers face many challenges following initiation of ART . The development of jaundice is uncommon but worrisome . In this case, two distinct and contrasting episodes of jaundice were observed . In the first instance, isolated elevation of the indirect bilirubin without elevation of the alkaline phosphatase was noted . The normal PT and serum aminotransferase levels indicate the absence of intrinsic liver dysfunction . Elevations in the indirect bilirubin may result from either impaired uptake/conjugation or excess production . The latter, usually from acquired hemolysis, may be a complication of an occult NHL . A work-up for this AIDS-related malignancy was not initiated since the caregivers recognized jaundice as a complication of IDV, which inhibits UDP-glucuronyl transferase and produces a Gilbert's-like syndrome . Physicians can expect to encounter this syndrome even more frequently with ATV . Experienced patients given RTV-boosted ATV have experienced elevations of unconjugated hyper-bilirubinemia in up to 45 percent of cases in clinical trials . However, such elevations do not reflect liver dysfunction and symptomatic jaundice requiring dosage reduction that occurred infrequently (7 to 8 percent of study patients) . Counseling patients about this syndrome may promote adherence and prevent self-directed interruptions of ATV that compromise efficacy . The second case of jaundice provides a more formidable diagnostic challenge . The triad of LFT abnormalities (mild elevation of aminotransferases, normal PT, and marked cholestatic jaundice) implies an acute process that is mildly toxic to hepatocytes without affecting their synthetic function . The subacute nature of the patient's cholestatic jaundice suggests either intrahepatic infiltrative disease of the liver or extrahepatic obstruction of the biliary tree, most likely due to the patient's relatively modest level of pain and lack of fever . Despite LFT abnormalities occurring 17 months after a switch in his ART, cumulative drug-related toxicities must still be considered . Ritonavir can produce significant elevations in the AST/ALT, especially with pre-existing chronic liver disease as with hepatitis C virus coinfection . The NRTIs can produce hepatic steatosis, a result of mitochondrial toxicity and impaired fatty acid oxidation . However, jaundice and cholestasis are not typical of the latter syndrome . With a negative contrast CT that excludes parenchymal liver disease, investigation of the biliary tree to assess the presence of AIDS-related cholangitis was the next step . Performing a sphincterotomy or stent placement, and obtaining brushings or biopsy specimens to determine the extent of extrahepatic obstruction may help define a pathogen and be life-saving . The negative results of the ERCP justify the final diagnostic step, a liver biopsy to evaluate microscopic infiltrative disease that might not have been detected on contrast abdominal CT . Examples might include granulomatous disease (MAC), fungal etiologies (histoplasmosis), carcinomatosis (lymphoma, hepatoma, cholangiocarcinoma), and microvascular disease (bacillary angiomatosis) . The failure to observe granulomatous inflammation in the liver does not exclude MAC infection, as MAC may involve other peri-aortic or mesenteric lymph nodes . This form of IRIS is unlikely given the abdominal CT findings, lack of systemic complaints, and extended persistence of liver aminotransferases . The nonspecific results of the liver biopsy are a common outcome in advanced AIDS patients with elevated alkaline phosphatase levels . Despite not having identified a pathogen, the biopsy establishes chronic liver disease and prompts re-evaluation and change of treatment to NFV . The subsequent normalization of the patient's aminotransferase levels suggests a prior adverse effect of LPV/r in the setting of unexplained, chronic liver disease . Most importantly, this case highlights the importance of HIV caregivers to review ART for safety when noting chronic liver dysfunction . Patients need to be counseled to minimize acetaminophen use, to consume alcohol in moderation, and to avoid behavior with risk for hepatitis C . Finally, all HIV patients should receive appropriate vaccination against hepatitis A and B if serology shows lack of protective immunity. J Ind Microbiol Biotechnol, 2004 Feb, 31(2), 83 - 7 Epub 2004 Feb 19. Compatibility of alkaline xylanases from an alkaliphilic Bacillus NCL (87-6-10) with commercial detergents and proteases; Kamal Kumar B et al.; Alkaline xylanases from alkaliphilic Bacillus strains NCL (87-6-10) and Sam III were compared with the commercial xylanases Pulpzyme HC and Biopulp for their compatibility with detergents and proteases for laundry applications . Among the four xylanases evaluated, the enzyme from the alkaliphilic Bacillus strain NCL (87-6-10) was the most compatible . The enzyme retained its full activity (40 degrees C for 1 h) in the presence of detergents, whereas Pulpzyme HC and Sam III showed only 30% and 50% of their initial activity, respectively . Biopulp, though stable to detergents, had only marginal activity (5%)at pH 10 . However, all four enzymes retained significant activity (80%) for 60 min in the presence of the proteases Alcalase and Conidiobolus protease . Supplementation of the enzyme enhanced the cleaning ability of the detergents. Protein Eng Des Sel, 2004 Jan, 17(1), 85 - 93 Fusion of the antiferritin antibody VL domain to barnase results in enhanced solubility and altered pH stability; Martsev SP et al.; Chimeric immunotoxins that combine antigen recognition domains of antibodies and cytotoxic RNases have attracted much attention in recent years as potential targeted agents for cancer immunotherapy . In an attempt to obtain a structurally minimized immunofusion for folding/stability studies, we constructed the chimeric protein VL-barnase . The chimera comprises a small cytotoxic enzyme barnase, ribonuclease from Bacillus amyloliquefaciens, fused to the C-terminus of the light chain variable domain (VL) of the anti-human ferritin monoclonal antibody F11 . While the individual VL domain was expressed in Escherichia coli as insoluble protein packed into inclusion bodies, its fusion to barnase resulted in a significant ( approximately 70%) fraction of soluble protein, with only a minor insoluble fraction ( approximately 30%) packed into inclusion bodies . The in vivo solubilizing effect of barnase was also observed in vitro and suggests a chaperone-like role that barnase exerted with regard to the N-terminal VL domain . Cytoplasmic VL-barnase was analyzed for structural and functional properties . The dimeric state of the chimeric protein was demonstrated by size-exclusion chromatography, thus indicating that fusion to barnase did not abrogate the intrinsic dimerization propensity of the VL domain . Ferritin-binding affinity and specificity in terms of constants of association with isoferritins were identical for the isolated VL domain and its barnase fusion, and RNase activity remained unchanged after the fusion . Intrinsic fluorescence spectra showed a fully compact tertiary structure of the fusion protein . However, significantly altered pH stability of the fusion protein versus individual VL and barnase was shown by the pH-induced changes in both intrinsic fluorescence and binding of ANS . Together, the results indicate that VL-barnase retained the antigen-binding affinity, specificity and RNase activity pertinent to the two individual constituents, and that their fusion into a single-chain chimeric protein resulted in an altered tertiary fold and pH stability. Arch Biochem Biophys, 2004 Jan 15, 421(2), 227 - 35 Antiamylase-pullulanase enzyme monoclonals which specifically inhibit amylase or pullulanase activity; Kim CH et al.; Monoclonal antibodies against amylase-pullulanase enzyme from Bacillus circulans F-2 have been produced to locate and characterize the catalytic sites of the enzyme . The antibodies have been examined for inhibition of both enzyme activities of amylase and pullulanase and then classified into four types: Type I which inhibited amylase activity, Type II which inhibited pullulanase activity, Type III which inhibited both enzyme activities, and Type IV which had no effect on either enzyme activity . Only two monoclonal antibodies (MAP-12 and MAP-17) as Type I and two antibodies (MAP-3 and MAP-5) as Type II were isolated . The inhibitory activities of the antibodies were characterized and compared . In Type II antibodies, the maximal demonstrated inhibition on the pullulanase activity was 88% for MAP-3 with 1 microg of antibody and 90% for MAP-5 with 2 microg of antibody, but did not inhibit the amylase activity . In Type I antibodies, in contrast, the maximal demonstrated inhibition on the amylase activity was 94% for MAP-12 and 97% for MAP-17 with 1 microg of antibody, respectively, but no inhibition of the pullulanase was noted . MAP-12 recognized sequential epitope, while MAP-17 recognized conformation-dependent epitope of amylase activity-related regions . However, both MAP-3 and MAP-5 recognized the conformation-dependent epitope of the pullulanase activity-related region . Furthermore, the antibodies of MAP-3, MAP-5, MAP-12, and MAP-17 did not compete with one another for binding to the enzyme, indicating that they have different target epitopes on the enzyme . Antibody binding of MAP-12 and MAP-17 to the enzyme was not specifically affected by any of the antiamylase compounds tested: (a) nojirimycin; and (b) 1-deoxynojirimycin . Kinetic analysis of their effects provides evidence that both antibodies of MAP-12 and MAP-17 decrease the catalytic rate of enzyme activity and have little or no effect on substrate binding. Antimicrob Agents Chemother, 2004 Mar, 48(3), 1058 - 60 Inhibition of beta-lactamase II of Bacillus cereus by penamaldic derivatives of penicillins; Navarro PG et al.; The penamaldic derivatives of amoxicillin, ampicillin, and penicillins G and V, stabilized with Zn(2+), were obtained from a methanolic medium . The enzymatic kinetic results show that the these derivatives elicit reversible inhibition of the enzyme metallo-beta-lactamase from Bacillus cereus, with inhibition constant values determined at pH 7.0 and 25 degrees C. Semin Oncol, 2004 Feb, 31(1 Suppl 1), 112 - 6 Vaccine therapy for small cell lung cancer; Krug LM; One novel approach to the treatment of lethal residual disease in patients with small cell lung cancer (SCLC) relies on the induction of a host-immune response to attack chemoresistant tumor cells . Because of its neuroectodermal origin, SCLC has a number of specific antigens that could be capitalized on as immune targets . This article reviews two vaccine strategies currently in clinical study . The anti-idiotype vaccine to the GD3 ganglioside, BEC-2, has recently been tested in a phase III trial . In this trial, patients with SCLC who had completed initial chemotherapy were randomized to observation or vaccination with BEC-2 plus bacillus Calmette-Guerin adjuvant . A series of other trials have established the immunogenicity of several keyhole limpet hemocyanin conjugate vaccines relevant to SCLC, including GM2, Globo H, fucosyl GM1, and polysialic acid . To optimize an immune response against a broad range of tumor phenotypes, these components will be combined into a polyvalent vaccine . A randomized phase II trial of this polyvalent vaccine is planned to start in 2004. Rev Med Liege, 2003 Dec, 58(12), 757 - 60 {Gallbladder tuberculosis associated with cholelithiasis}; Rouas L et al.; Tuberculosis of the gallbladder is rare, even in our country known for being an endemic area . The positive diagnosis depends on suspicion of tuberculosis, peroperative findings and histological examination . From a review of the literature, the physiopathology of this infection is discussed, emphasizing the role of lithiasis in the development of tuberculous lesions . The authors report a case of gallbladder tuberculosis in a female patient who presented with a clinical picture of chronic cholelithiasis . The diagnosis of gallbladder tuberculosis was reached only after surgery and proven by histopathology . In our case, the presence of stones associated with non specific inflammatory alterations and possibly low resistance against tubercle bacillus, is believed to have been of importance for the development of the tuberculous infection. Proc Natl Acad Sci U S A, 2004 Mar 2, 101(9), 2696 - 9 Epub 2004 Feb 20. Induction and transmission of Bacillus thuringiensis tolerance in the flour moth Ephestia kuehniella; Rahman MM et al.; The use of Bacillus thuringiensis (Bt) endotoxins to control insect vectors of human diseases and agricultural pests is threatened by the possible evolution of resistance in major pest species . In addition to high levels of resistance produced by receptor insensitivity (5, 16, 17), several cases of tolerance to low to medium levels of toxin have been reported in laboratory colonies of lepidopteran species (3, 18) . Because the molecular basis of some of these cases of tolerance to the toxin are not known, we explored alternative mechanisms . Here, we present evidence that tolerance to a Bt formulation in a laboratory colony of the flour moth Ephestia kuehniella can be induced by preexposure to a low concentration of the Bt formulation and that the tolerance correlates with an elevated immune response . The data also indicate that both immune induction and Bt tolerance can be transmitted to offspring by a maternal effect and that their magnitudes are determined by more than one gene. J Immunol, 2004 Mar 1, 172(5), 2935 - 43 Posttranscriptional inhibition of gene expression by Mycobacterium tuberculosis offsets transcriptional synergism with IFN-gamma and posttranscriptional up-regulation by IFN-gamma; Qiao Y et al.; Host defense against Mycobacterium tuberculosis requires the cytokine IFN-gamma and IFN regulatory factor 1 (IRF-1), a transcription factor that is induced to high levels by IFN-gamma . Therefore, we chose to study regulation of IRF-1 expression as a model for effects of M . tuberculosis on response to IFN-gamma . We found that IRF-1 mRNA abundance increased far more than transcription rate in human monocytic THP-1 cells stimulated by IFN-gamma, but less than transcription rate in cells infected by M . tuberculosis . IFN-gamma stimulation of infected cells caused a synergistic increase in IRF-1 transcription, yet IRF-1 mRNA abundance was similar in uninfected and infected cells stimulated by IFN-gamma, as was the IRF-1 protein level . Comparable infection by Mycobacterium bovis bacillus Calmette-Guerin failed to induce IRF-1 expression and had no effect on the response to IFN-gamma . We also examined the kinetics of transcription, the mRNA t(1/2), and the distribution of IRF-1 transcripts among total nuclear RNA, poly(A) nuclear RNA, and poly(A) cytoplasmic RNA pools in cells that were infected by M . tuberculosis and/or stimulated by IFN-gamma . Our data suggest that infection by M . tuberculosis inhibits RNA export from the nucleus . Moreover, the results indicate that regulated entry of nascent transcripts into the pool of total nuclear RNA affects IRF-1 expression and that this process is stimulated by IFN-gamma and inhibited by M . tuberculosis . The ability of infection by M . tuberculosis to limit the increase in IRF-1 mRNA expression that typically follows transcriptional synergism may contribute to the pathogenicity of M . tuberculosis. Infect Immun, 2004 Mar, 72(3), 1504 - 11 Gammadelta T cells in immunity induced by Mycobacterium bovis bacillus Calmette-Guérin vaccination; Lee J et al.; Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccination is efficacious for newborns or adults with no previous exposure to environmental mycobacteria . To determine the relative contribution and the nature of gammadelta T-cell receptor-positive T cells in newborns, compared to CD4(+) T cells, in immunity induced by M . bovis BCG vaccination, 4-week-old specific-pathogen-free pigs were vaccinated with M . bovis BCG and monitored by following the gammadelta T-cell immune responses . A flow cytometry-based proliferation assay and intracellular staining for gamma interferon (IFN-gamma) were used to examine gammadelta T-cell responses . Pigs were found to mount Th1-like responses to M . bovis BCG vaccination as determined by immunoproliferation and IFN-gamma production . The gammadelta T-cell lymphoproliferation and IFN-gamma production to stimulation with mycobacterial antigens were significantly enhanced by M . bovis BCG vaccination . The relative number of proliferating gammadelta T cells after stimulating peripheral blood mononuclear cells with Mycobacterium tuberculosis H37Rv culture filtrate protein was higher than that of CD4(+) T cells at an early time point after M . bovis BCG vaccination, but CD4(+) T cells were found to be more abundant at a later time point . Although the gammadelta T-cell responses were dependent on the presence of CD4(+) T cells for the cytokine interleukin-2, the enhanced gammadelta T cells were due to the intrinsic changes of gammadelta T cells caused by M . bovis BCG vaccination rather than being due solely to help from CD4(+) T cells . Our study shows that gammadelta T cells from pigs at early ages are functionally enhanced by M . bovis BCG vaccination and suggests an important role for this T-cell subset in acquired immunity conferred by M . bovis BCG vaccination. Biometals, 2004 Feb, 17(1), 45 - 52 Qualitative and quantitative composition of pigments in Phaeodactylum tricornutum (Bacillariophyceae) stressed by iron; Kosakowska A et al.; The effect of Fe(III) deficiency on qualitative and quantitative changes in pigment composition in Phaeodactylum tricornutum Bohlin was demonstrated by HPLC and AAS . Maximum content of pigments showed the diatom cells incubated at the optimum iron concentration, i.e., 10 microM . The contents of chlorophyll a, chlorophyll c1 + c2, fucoxanthin, diadinoxanthin and beta,beta-carotene were 109.99, 20.16, 40.39, 1.29 and 1.48 fg per cell, respectively . The results obtained showed that Fe(III) affected qualitative and quantitative pigment composition in P . tricornutum . The content of individual pigments, proportions between accompanying pigments and their ratios to chlorophyll a were important indicators of phytoplankton response to iron stress . The strong reduction in beta,beta-carotene content, several times (2-5) increase in diadinoxanthin level as compared to beta,beta-carotene, and high amount of diadinoxanthin in relation to chlorophyll a were observed in algae growing at very low Fe(III) concentrations, 0.001 and 0.01 microM . The data suggested that phytoplankton pigments could be a potential physiological marker. J Econ Entomol, 2003 Dec, 96(6), 1838 - 42 Geographic variation in susceptibility of Chilo suppressalis (Lepidoptera: Pyralidae) to Bacillus thuringiensis toxins in China; Meng F et al.; Geographic variation in the susceptibility of the striped stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), in China to Bacillus thuringiensis (Bt) insecticidal crystal proteins Cry1Ac and Cry1Ab was studied to establish baseline information for comparing the future response of populations with increased exposure to Bt products . Rice is the major host of C . suppressalis, and Bt rice ma) be released in China in the near future . Twelve populations of the pest were collected from the major rice-growing regions of China . LC50 estimates were determined for all populations for Cry1Ac and for eight populations for Cry1Ab . The bioassay results indicated that the range of LC50 in neonate larvae to Cry1Ac and Cry1Ab was from approximately 15 to approximately 157 mg (AI)/L and approximately 2 to approximately 34 mg (AI)/L, respectively . LC50 values were lower for Cry1Ab than for Cry1Ac, and there was a significant positive correlation between the two toxins tested. J Econ Entomol, 2003 Dec, 96(6), 1786 - 91 Efficacy of VectoBac (Bacillus thuringiensis variety israelensis) formulations for mosquito control in Australia; Russell TL et al.; Laboratory bioassays were conducted on the efficacy of a water-dispersible granule (WG) formulation of Bacillus thuringiensis variety israelensis (VectoBac WG; active ingredient {AI}: 3,000 Bti international toxic units {ITU}/mg) against third instars of six common Australian mosquito species, Aedes aegypti (L.), Ochlerotatus vigilax (Skuse), Ochlerotatus notoscriptus (Skuse), Culex sitiens Wiedemann, Culex annulirostris Skuse, and Culex quinquefasciatus Say . The normal model for log-linear mortality data was used to determine laboratory 48-h LC50 and LC95 values . The target mosquito species tested were extremely sensitive to the VectoBac WG formulation, with the most sensitive species (Cx . annulirostris and Cx . quinquefasciatus, LC95 value of 0.019 ppm) being twice as susceptible as the most tolerant (Oc . notoscriptus, LC95 value of 0.037 ppm) . Cx . annulirostris was selected as a target species for a small-plot evaluation of VectoBac WG and VectoBac 12 aqueous solution (AS) ({AI}: 1,200 Bti ITU/mg) efficacy over time, in freshwater in southeastern Queensland, Australia . Replicated cohorts of caged third instars were exposed weekly to six concentrations of WG formulation (0.004-0.13 ppm) and three concentrations of the 12AS formulation (0.04-0.13 ppm) . In water with high organic content, treatment concentrations of 0.008 ppm WG and 0.04 ppm 12AS and above produced significant larval control (> or = 96%) at 48 h posttreatment, with no residual control at week 1 . Water quality was not affected by treatment with either formulation. J Econ Entomol, 2003 Dec, 96(6), 1662 - 7 Increased mortality of gypsy moth Lymantria dispar (L.) (Lepidoptera: Lymantriidae) exposed to gypsy moth nuclear polyhedrosis virus in combination with the phenolic gycoside salicin; Cook SP et al.; Second instar gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae), larvae suffered significantly greater mortality from aerially applied gypsy moth nuclear polyhedrosis virus (Gypchek) when the virus was consumed on quaking aspen, Populus tremuloides Michx., versus red oak, Quercus spp . L., foliage . Laboratory assays in which various doses of Gypchek and salicin (a phenolic glycoside present in aspen foliage) were tested in combination demonstrated that salicin significantly increased total larval mortality and lowered the LD50 estimates (dose of Gypchek that resulted in 50% population mortality) for the virus, although not significantly . While salicin did not impact larval survival in the absence of Gypcek, it did act to significantly deter feeding when it was present in high concentrations (up to 5.0%) within the treatment formulations . The enhanced activity of Gypchek in the presence of salicin is similar to prior reports of enhanced activity of the bacterial pathogen Bacillus thuringiensis when consumed concurrently with phenolic glycosides commonly present in aspen foliage . The enhancement of viral activity is in contrast to the inhibitory effects on the virus reported for another common group of phenolic compounds, tannins. J Econ Entomol, 2003 Dec, 96(6), 1653 - 61 Evaluation of oils and microbial pathogens for control of lepidopteran pests of sweet corn in New England; Hazzard RV et al.; Vegetable and mineral oil, Beauveria bassiana (Balsamo) and Bacillus thuringiensis subsp . kurstaki Berliner were evaluated for control of Ostrinia nubilalis (Hubner) (Lepidoptera: Pyralidae), Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) and Spodoptera frugiperda (J . E . Smith) (Lepidoptera: Noctuidae) in sweet corn (Zea mays L.) . Field experiments in Maine and Massachusetts during 1993 and 1994 evaluated oils and pathogens singly or in combinations, using a single application directly to the top of the silk channel, immediately after pollination . Mineral oil alone provided equal (1993) or better (1994) control compared with corn oil . In both years, mineral or corn oil plus B . thuringiensis resulted in 93-98% marketable ears, compared with 48-52% marketable ears in untreated plots . In three factorial experiments with B . bassiana, B . thuringiensis and corn oil, B . bassiana at 5 x 10(7) conidia per ear provided little or no control while B . thuringiensis and corn oil provided significant though not always consistent control of all three species . The combination of B . thuringiensis and corn oil provided the largest and most consistent reduction in numbers of larvae and feeding damage to ears. Insect Biochem Mol Biol, 2004 Jan, 34(1), 101 - 12 Analysis of glycan structures on the 120 kDa aminopeptidase N of Manduca sexta and their interactions with Bacillus thuringiensis Cry1Ac toxin; Knight PJ et al.; The Bacillus thuringiensis Cry1Ac toxin specifically binds to a 120 kDa aminopeptidase N (APN) receptor in Manduca sexta . The binding interaction is mediated by GalNAc, presumably covalently attached to the APN as part of an undefined glycan structure . Here we detail a simple, rapid and specific chemical deglycosylation technique, applicable to glycoproteins immobilized on Western blots . We used the technique to directly and unambiguously demonstrate that carbohydrates attached to 120 kDA APN are in fact binding epitopes for Cry1Ac toxin . This technique is generally applicable to all putative Cry toxin/receptor combinations . We analyzed the various glycans on the 120 kDA APN using carbohydrate compositional analysis and lectin binding . The data indicate that in the average APN molecule, 2 of 4 possible N-glycosylation sites are occupied with fucosylated paucimannose {Man(2-3)(Fuc(1-2)GlcNAc(2)-peptide} type N-glycans . Additionally, we identified 13 probable O-glycosylation sites, 10 of which are located in the Thr/Pro rich C-terminal "stalk" region of the protein . It is likely that 5-6 of the 13 sites are occupied, probably with simple {GalNAc-peptide} type O-glycans . This O-glycosylated C-terminal stalk, being GalNAc-rich, is the most likely binding site for Cry1Ac. J Infect Dis, 2004 Mar 1, 189(5), 812 - 9 Epub 2004 Feb 13. Mapping immune reactivity toward Rv2653 and Rv2654: two novel low-molecular-mass antigens found specifically in the Mycobacterium tuberculosis complex; Aagaard C et al.; New tools are urgently needed for the detection of latent tuberculosis (TB) . We evaluated the diagnostic potential of 2 novel Mycobacterium tuberculosis complex-specific candidate antigens (Rv2653 and Rv2654) and investigated T cell recognition during natural infection in humans and experimental infection in guinea pigs . Peripheral blood mononuclear cells stimulated with peptide pools covering the full length of Rv2654 induced interferon- gamma release in 10 of 19 patients with TB . Neither Rv2654 single peptides nor Rv2654 pools were recognized by bacille Calmette-Guerin-vaccinated donors . However, peptides from Rv2653 were recognized by both patients group . The cross-reactive epitope(s) in Rv2653 were located in a 36-amino acid stretch in the center of the molecule . Rv2654 also induced M . tuberculosis-specific skin-test responses in 3 of 4 aerosol-infected guinea pigs . Rv2654 is a strongly recognized T cell antigen that is highly specific for TB and has potential as a novel cell-mediated immunity-based TB diagnostic agent. Int J Tuberc Lung Dis, 2004 Jan, 8(1), 45 - 51 Tuberculin reactivity and tuberculosis epidemiology in the Pakaanóva (Wari') Indians of Rondônia, south-western Brazilian Amazon; Escobar AL et al.; OBJECTIVE: To investigate the characteristics of tuberculin skin test reactivity in the Pakaanova Indians, in Amazonia, Brazil, after revaccination of all study participants with bacille Calmette-Guerin (BCG) . METHODS: The investigation was designed as a post-BCG vaccination purified protein derivative (PPD) survey . Data included PPD readings, age, sex, nutritional status, place of residence, previous tuberculosis, physical examinations and BCG status . Bivariate and multivariate logistic regression analyses were conducted . RESULTS: About 90% (n = 505) of the total population participated . One third (32.1%) of the subjects presented induration > or = 10 mm at 72 h . Induration sizes showed weak linear correlation with age; differences between sexes were not observed . Skin reaction was not associated with nutritional status . Individuals with a history of tuberculosis were six times more likely to test positive . History of tuberculosis, age, and previous BCG vaccination were significantly associated with PPD reactivity in the multivariate analyses . CONCLUSION: The Pakaanova showed a high proportion (58.4%) of non-reactors, even with a recent BCG booster . Sex differences in PPD reactivity were either not present or could not be demonstrated . The association between age and PPD reactivity resembles that observed in other Amazonian populations . The authors discuss the potential of PPD testing as a screening tool to enhance tuberculosis detection, especially in indigenous populations in Amazonia with limited access to health services. Crit Care Nurs Q, 2004 Jan-Mar, 27(1), 87 - 91 A case study in Hansen's disease acquired after heart transplant; Launius BK et al.; Hansen's disease, leprosy, is a chronic infectious disease caused by the acid-fast bacillus Mycobacterium leprae . There are multiple forms of the disease ranging from the relatively benign to the progressive, malignant lepromatous leprosy . There is effective antimicrobial treatment available that is capable of curing the disease . We report the case of a post heart transplant patient acquiring Hansen's disease. Cancer Gene Ther, 2004 Mar, 11(3), 194 - 207 Treatment of bladder carcinomas using recombinant BCG DNA vaccines and electroporative gene immunotherapy; Lee CF et al.; Intravesical immunotherapy with live Mycobacterium bovis bacillus Calmette-Guerin (BCG) is the treatment of choice for superficial bladder cancers . Nevertheless, a significant proportion of patients do not respond to this therapy, and adverse effects are common . Here, we report the cloning of recombinant mycobacterial DNA vaccines and demonstrate the ability of multicomponent and multisubunit DNA vaccines to enhance Th1-polarized cytokine-mediated responses as well as effector cell responses . Splenocytes from immunized groups of mice were restimulated in vitro and examined for cytotoxicity against murine bladder tumur (MBT-2) cells . We used four combined recombinant BCG DNA vaccines (poly-rBCG) for electroporative gene immunotherapy (EPGIT) in vivo, and found that tumor growth was significantly inhibited and mouse survival was prolonged . Increased immune cell infiltration and induction of apoptosis were noted after treatment with poly-rBCG alone, with the murine interleukin-12 (mIL-12) vaccine alone, and-most significantly-with the poly-rBCG+mIL-12 vaccine combination . Electroporation of poly-rBCG+mIL-12 resulted in complete tumor eradication in seven of eight mice (P<.01) within 28 days . Thus, EPGIT using multicomponent multisubunit BCG is highly effective in the treatment of bladder cancer . This approach presents new possibilities for the treatment of bladder cancer using recombinant BCG DNA vaccines. J Bacteriol, 2004 Mar, 186(5), 1229 - 38 Plasmid-dependent methylotrophy in thermotolerant Bacillus methanolicus; Brautaset T et al.; Bacillus methanolicus can efficiently utilize methanol as a sole carbon source and has an optimum growth temperature of 50 degrees C . With the exception of mannitol, no sugars have been reported to support rapid growth of this organism, which is classified as a restrictive methylotroph . Here we describe the DNA sequence and characterization of a 19,167-bp circular plasmid, designated pBM19, isolated from B . methanolicus MGA3 . Sequence analysis of pBM19 demonstrated the presence of the methanol dehydrogenase gene, mdh, which is crucial for methanol consumption in this bacterium . In addition, five genes (pfk, encoding phosphofructokinase; rpe, encoding ribulose-5-phosphate 3-epimerase; tkt, encoding transketolase; glpX, encoding fructose-1,6-bisphosphatase; and fba, encoding fructose-1,6-bisphosphate aldolase) with deduced roles in methanol assimilation via the ribulose monophosphate pathway are encoded by pBM19 . A shuttle vector, pTB1.9, harboring the pBM19 minimal replicon (repB and ori) was constructed and used to transform MGA3 . Analysis of the resulting recombinant strain demonstrated that it was cured of pBM19 and was not able to grow on methanol . A pTB1.9 derivative harboring the complete mdh gene could not restore growth on methanol when it was introduced into the pBM19-cured strain, suggesting that additional pBM19 genes are required for consumption of this carbon source . Screening of 13 thermotolerant B . methanolicus wild-type strains showed that they all harbor plasmids similar to pBM19, and this is the first report describing plasmid-linked methylotrophy in any microorganism . Our findings should have an effect on future genetic manipulations of this organism, and they contribute to a new understanding of the biology of methylotrophs. Onderstepoort J Vet Res, 2003 Dec, 70(4), 307 - 16 Review of blackfly (Diptera: Simuliidae) control in South Africa; Myburgh E et al.; The medical, veterinary and economic importance of blackflies in South Africa, and the historical development of blackfly control programmes in various South African rivers, are reviewed in this paper . In 1996 it was estimated that blackflies can cause more than R 88 million damages per annum along the middle and lower Orange River where Simulium chutteri is considered the main pest species . A clear link between the construction of dams and the spread of the blackfly problem was shown . Four phases characterize the development of blackfly control in South Africa: (1) during the 1960s blackflies in the Vaal River were controlled with DDT; (2), during the 1970s and into the 1980s blackflies were controlled using water-flow manipulation; (3) when used at strategic times, water-flow manipulation could be used to enhance the effect of natural predator populations; and (4) during the 1990s the organophosphate temephos and toxins produced by the bacterium Bacillus thuringiensis var . israelensis were tested for their efficacy against blackflies . The larvicides temephos and B . thuringiensis proved to be effective and are still used in several control programmes . The latest research focuses on the factors that influence adult blackfly survival and annoyance, as well as the development of methods that can be used to protect sheep from blackfly attacks. Onderstepoort J Vet Res, 2003 Dec, 70(4), 255 - 63 Leishmania donovani-derived lipophosphoglycan plus BCG induces a Th1 type immune response but does not protect Syrian golden hamsters (Mesocricetus auratus) and BALB/c mice against Leishmania donovani; Tonui WK et al.; The efficacy of Leishmania donovani-derived lipophosphoglycan (LPG) plus Mycobacterium bovis Bacille Calmette-Guerin (BCG) as a vaccine candidate against visceral leishmaniosis in susceptible BALB/c mouse and Syrian golden hamster (Mesocricetus auratus) models was investigated . Following a triple vaccination with a total dose of 150 microl BCG plus 60 microg or 30 microg of LPG for hamsters and BALB/c mice respectively, there were no noticeable side effects both locally and systemically; implying that the molecule was safe at this dosage level . Vaccinated animals demonstrated an activation of both the humoral as well as cell-mediated responses to LPG, which correlated with resistance against the disease . Protection by LPG plus BCG, was however, poor as the remaining immunized animals showed disease progression leading to severity of the disease as illustrated by emaciation, mass loss and heavy splenic parasitaemia in hamsters . These data nevertheless suggest that it may be rewarding to further evaluate the potential of LPG as a vaccine candidate in leishmaniosis using other adjuvants, which may enhance its immunogenicity. Pest Manag Sci, 2004 Feb, 60(2), 167 - 72 Long-term selection for resistance to transgenic cotton expressing Bacillus thuringiensis toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae); Meng F et al.; Selection experiments for resistance to transgenic Bt cotton expressing Cry1Ac toxin in Helicoverpa armigera (Hubner) were conducted using a leaf-feeding method with 42 selection episodes over 45 generations . The cotton bollworm developed resistance to transgenic Bt cotton after 12 generations (F12) of selection . The survival rate of F12 neonates feeding on leaves of seedling stage for 4 days and boll-opening stage for 5 days of R19 line were ca 34 and 72%, respectively, compared with ca 0 and 40% for a non-selected sister strain (NYCS), but lower than or similar to that of F12 feeding on leaves of non-Bt cotton, Sumian 12 (ca 87 or 76%) . Resistance to B thuringiensis HD-1 Dipel in neonates (F12) was ca 6-fold . After 42 generations of selection, the strain developed a very high level of resistance to Cry1Ac protoxin, 210 g kg(-1) MVPII wettable powder and 200 g litre(-1) MVPII liquid formulation, the resistance ratios being 1680-, 1780- and ca 1200-fold, respectively, compared with a laboratory susceptible strain (HZS) . When compared with the non-selected NYCS, the resistance ratios to the above toxins were ca 540-, 580- and 510-fold, respectively, but to Dipel only ca 16-fold . The results indicated that it is very important to develop and implement effective resistance-management strategies and to detect early resistance to Bt cotton in field populations. J Food Prot, 2004 Feb, 67(2), 399 - 402 Factors contributing to the occurrence of antimicrobial drug residues in Kenyan milk; Shitandi A et al.; The study investigated factors contributing to the occurrence of antimicrobial drug residues in milk within four major milk production districts in Kenya . The frequency of contamination was studied among small- and large-scale dairy producers to determine if there were differences between the two types of producers . Field samples (n = 1,600) were analyzed with the improved Dutch tube diffusion test, a microbial inhibitor test (Bacillus stearothermophilus) . In total, 144 and 64 samples from small- and large-scale producers, respectively, were found to contain beta-lactam antibiotics at levels exceeding the established Codex maximum residue level for penicillin G (4 microg/kg) . The difference in results between the two categories of producers was found to be significant (P < 0.001) . To explain the higher frequency of antibiotic contamination of milk from small-scale producers, a questionnaire was constructed and used with 220 randomly selected smallholders in the selected districts . The results suggested (i) lack of understanding of risks related to antibiotic contamination of food, (ii) poor or no treatment records, and (iii) lack of a monitoring system as major risks for contamination . It was concluded that intensification of the education among small-scale dairy producers would greatly reduce the occurrence of antimicrobial residues in milk. J Food Prot, 2004 Feb, 67(2), 387 - 90 A colony blot immunoassay for the rapid identification of Bacillus cereus; Chen CH et al.; A colony blot immunoassay was developed for the rapid identification of Bacillus cereus using antibodies against the 28.5-kDa cell-surface antigen of B . cereus . Suspect colonies from plates were blotted onto a Whatman #541 membrane, dried, and fixed by UV irradiation . The membrane was then immersed in an anti-B . cereus antibody-horseradish peroxidase conjugate for 60 min . After washing and reacting with 4-chloro-1-naphthol and H2O2, the appearance of purple spots indicated the presence of B . cereus . This assay effectively identified 61 of 62 B . cereus strains tested . Among 38 non-B . cereus strains, which were other Bacillus spp . (19 genera), 36 gave true-negative results, and 2 showed false-positive results . The sensitivity and specificity for B . cereus were 98.4 and 94.7%, respectively . The present assay is easy to use, and the rapid identification of B . cereus can be completed in 2.5 h. Rev Chir Orthop Reparatrice Appar Mot, 2004 Feb, 90(1), 75 - 8 {Muscular tuberculosis: an unusual focus of Koch bacillus}; Dendane A et al.; We report an exceptional case of tuberculosis in an 8-year-old girl . The focus of infection was situated in the anterior muscle compartment of the left thigh . Diagnosis was established on the basis of histological findings . A 12-month antituberculosis regimen was given . Outcome was good with no general or functional sequelae . Based on this case and data reported in the literature, we discuss the routes of dissemination of the Koch bacillus explaining this unusual sporadic localization. Biochemistry, 2004 Feb 24, 43(7), 2080 - 90 Phosphatidylinositol-specific phospholipase C forms different complexes with monodisperse and micellar phosphatidylcholine; Berg OG et al.; Phosphatidylinositol-specific phospholipase C (PI-PLC) from Bacillus cereus forms a premicellar complex E(#) with monodisperse diheptanoylphosphatidylcholine (DC(7)PC) that is distinguishable from the E complex formed with micelles . Results are interpreted with the assumption that in both cases amphiphiles bind to the interfacial binding surface (i-face) of PI-PLC but not to the active site . Isothermal calorimetry and fluorescence titration results for the binding of monodisperse DC(7)PC give an apparent dissociation constant of K(2) = 0.2 mM with Hill coefficient of 2 . The gel-permeation, spectroscopic, and probe partitioning behaviors of E(#) are distinct from those of the E complex . The aggregation and partitioning behaviors suggest that the acyl chains in E(#) but not in E remain exposed to the aqueous phase . The free (E) and complexed (E(#) and E) forms of PI-PLC, each with distinct spectroscopic signatures, readily equilibrate with changing DC(7)PC concentration . The underlying equilibria are modeled and their significance for the states of the PI-PLC under monomer kinetic conditions is discussed to suggest that the Michaelis-Menten complex formed with monodisperse DC(7)PC is likely to be E(#)S or its aggregate rather than the classical monodisperse ES complex. Zhejiang Da Xue Xue Bao Yi Xue Ban, 2004 Jan, 33(1), 65 - 7 {Experimental study on attachment of bacillus Calmette-Guerin to bladder wall with different types of injury}; Ding GQ et al.; OBJECTIVE: To investigate whether different types of injury on bladder wall can influence bacillus Calmette-Guerin (BCG) attachment . METHODS: The bladder mucosa of 24 rabbits were treated by electrocautery,cryocautery and incision on left lateral wall, right lateral wall and posterior wall, respectively . Then radiolabeled BCG ((3)H-BCG) was instilled into bladder . Two hours latter, the injured bladder wall with different methods and non-injured wall (anterior wall of bladder) were surgically removed and digested . The quantity of BCG of each specimen was determined by liquid scintillation counter . RESULT: The quantity of BCG attachment to bladder wall with different injuries was significantly higher than that of non-injured wall (P<0.001), meanwhile there was no statistically difference among the BCG levels of different injury types (P>0.05) . CONCLUSION: BCG attachment is not influenced by different types of injury on the bladder wall. Appl Microbiol Biotechnol, 2004 Aug, 65(2), 158 - 62 Epub 2004 Feb 14. Influence of media and temperature on bacteriocin production by Bacillus cereus 8A during batch cultivation; Bizani D et al.; Cerein 8A is a bacteriocin produced by the soil bacterium Bacillus cereus 8A, isolated from native woodlands of Brazil . The influence of temperature and media on the growth of B . cereus 8A and the production of this bacteriocin was studied during batch cultivation . Maximum activity was detected by cultivation in brain/heart infusion broth, reaching 3200 activity units ml(-1) . Bacteriocin was also produced in peptone, MRS, Mueller-Hinton and nutrient broth, while no activity was observed during cultivation in thioglycollate or tryptic soy broth . Temperature had a strong influence on bacteriocin production, which was higher at 30 degrees C than at 25 degrees C . An important decrease in bacteriocin activity was observed at 37 degrees C . The relationship between growth and specific production rates, as a function of the temperature, showed different kinetics of production and there were several peaks in the specific production rates during growth . Bacteriocin was produced at the stationary phase, indicating it is synthesized as a secondary metabolite. Transplantation, 2004 Feb 15, 77(3), 456 - 59 Macrophage colony-stimulating factor inhibits tumor necrosis factor production and prolongs skin graft survival; Nishina T et al.; BACKGROUND: Despite the availability of a variety of immunosuppressive agents, acute rejection and infection after organ transplantation remain serious problems . METHODS AND RESULTS: We examined the effect of macrophage colony-stimulating factor (M-CSF) on the production of tumor necrosis factor (TNF) in a Bacille de Calmette Guerin-lipopolysaccharide-challenged mouse model . Both serial and repeated injections of M-CSF inhibited TNF production in a dose-dependent manner . Electrophoretic mobility shift assay showed that M-CSF-induced inhibition of TNF production was a result of suppression of nuclear factor-kappaB . High-dose M-CSF significantly prolonged skin graft survival in mice with orthotopic transplantation compared with the control and low-dose M-CSF groups . The combined administration of low-dose M-CSF and cyclosporine also significantly prolonged graft survival compared with the control and low-dose single agent-treated groups . CONCLUSIONS: Our results indicate that M-CSF at a high dose is a potent inhibitor of cytokine production and can potentially be used as an immunosuppressive agent for allograft rejection. J Egypt Soc Parasitol, 2003 Aug, 33(2), 517 - 30 Cytopathological action in mosquito larvae fed with Bacillus sphaericus (strain faiyoum) spore/crystal complex; Labib IM et al.; Ingestion of Bacillus sphaericus (strain faiyoum) spore/crystal complex by larvae of Anopheles pharoensis, Culex pipiens and Aedes caspius was rapidly followed by a dissolution of the protein crystalline inclusions inside the anterior stomach of the three species . During the first day of the, B . sphaericus spores germinated within the mid-gut lumen, and were in a vegetative stage between 36-48 hours after ingestion when larvae began to die . Ultrastructural observations focused on larval mid-gut showed alterations, which differ according to the mosquito species, being localized mainly in the gastric caecae and posterior stomach . With the bacterial concentration used, neither general cell swelling nor complete breakdown of the mid-gut epithelium was recorded before larval death . In An . pharoensis larval mid-gut epithelium large low-electron-density areas appeared, rough endoplasmic reticula formed numerous concentrical structures and mitochondria swelled . Large vacuoles (of unknown origin) appear early in the Cx . pipiens mid-gut cells and rough endoplasmic reticula broke into small vesicles . Mid-gut epithelial cells of Ae . caspius showed mitochondria swelling except in the anterior stomach and a vacuolization of smooth reticula: these aspects remained unchanged until the larvae died. J Egypt Soc Parasitol, 2003 Aug, 33(2), 425 - 36 Laboratory evaluation of Bacillus sphaericus recycling in mosquito larvae; Labib IM et al.; After ingestion by Culex pipiens and Anopheles pharoensis 4th instar larvae, spores of Bacillus sphaericus strain faiyoum rapidly germinated inside live mosquito midgut . Bacterial counts and electron microscopic observations on intoxicated larvae revealed that the number of viable spores rapidly decreased during the first 12 h, with a maximum between 12 and 24 h . In cadavers, the number of heat-resistant spores quickly increased between the first and second day post-feeding . After one week, the number of spores inside dead larvae reached approximately 20 times the number of ingested spores for both mosquito species (4 x 16(5) spores/larva) . Ultrathin sections of recycled spores showed the presence of a crystalline inclusion identical to that initially present in spores before ingestion . Bioassay on Cx pipiens 4th instar larvae showed a similar toxicity between in vivo recycled spores (LC50 = 1.1 +/- 0.3 x 10(5) spores/ml after 24h exposure) and culture-medium-grown spores of B . sphaericus strain faiyoum (LC50 = 1.7 +/- 10(5) spores/ml). J Egypt Soc Parasitol, 2003 Aug, 33(2), 331 - 40 Soil characteristics as factors governing the existence, recycling and persistence of Bacillus thuringiensis in Egypt; Merdan BA et al.; The simple correlation and regression analyses showed that the soil sand percentage and the available copper levels had significant negative and positive contributions, respectively, to the abundance and distribution of B . thuringiensis in the Egyptian agricultural soil . Among the B . thuringiensis isolates, only 11.26% of strains showed larvicidal activity against Culex pipiens . The larvicidal potency LD50 varied from 5.01X10(5) to 5.4X10(6) spores per milliliter among the isolates. New Microbiol, 2004 Jan, 27(1), 29 - 35 Population dynamics in ageing Helicobacter pylori; Cellini L et al.; The aim of this work was to characterize population changes occurring in aged broth cultures of Helicobacter pylori . Experiments were performed using clinical strains cultured immediately after isolation and after multiple subcultures in solid medium . Morphological changes in the ageing bacteria during a 7-day broth culture were analysed by optical and electron microscopy . The expression of the virulence factor, CagA, together with the presence of the cell cycle regulator, cGMP, were also assessed . The transition from bacillary to coccoid forms was the main morphological change observed in freshly isolated bacteria, together with the increase in cGMP from 1 to 2.25 nmoles/mg of proteins within the first 7 days of broth culture . A similar trend of morphological and physiological changes was observed in cells after multiple subcultures in solid medium with a major presence of large cell clusters . The cagA gene product was always expressed in all experimental conditions evaluated . These data show a significant morphological and physiological diversity in fresh, ageing and aged cultures of H . pylori. J Biol Chem, 2004 Apr 16, 279(16), 15779 - 86 Epub 2004 Feb 12. The role of Bacillus thuringiensis Cry1C and Cry1E separate structural domains in the interaction with Spodoptera littoralis gut epithelial cells; Avisar D et al.; The Bacillus thuringiensis delta-endotoxins Cry1C and Cry1E share toxicity against several important lepidopteran species . Their combined use to delay development of resistance in target insects depends on their differential interaction with the gut epithelial cells . The three structural domains and combinations of two consecutive domains of Cry1C and Cry1E were separately expressed in Escherichia coli, and their interactions with the brush border membrane vesicles (BBMV) of Cry1E-tolerant and -susceptible Spodoptera littoralis larvae were studied . About 80% reduction in binding of Cry1E and each of its separate domains to BBMV of Cry1E-tolerant larvae was observed, whereas Cry1C was toxic to all larvae and bound equally to BBMV derived from both Cry1E-tolerant and -susceptible larvae . These results suggest differential interactions of the two toxins with BBMV encompassing all three domains . Comparable binding assays performed with fluorescent Cry1C and Cry1C domain II showed that Cry1C has higher Bmax and lower Kd than Cry1C domain II and further supported the existence of toxin multisite interactions . Competitive binding assays were used to estimate the sequence of interaction events . Cry1C domain II could compete with domain III binding, whereas domain III did not interfere with domain II binding, indicating sequential interactions of domain III and then domain II with the same membrane site . No competition between domain II of Cry1C and Cry1E was observed, confirming the existence of different domain II binding sites for the two toxins . Taken together, all three domains specifically interact with the epithelial cell membrane . The folding of the three-domain toxin probably dictates the sequence of interaction events. Australas J Dermatol, 2004 Feb, 45(1), 51 - 4 Granulomas in common variable immunodeficiency: A diagnostic dilemma; Lun KR et al.; A 60-year-old man with common variable immunodeficiency presented with a 7-year history of violaceous plaques and papules on the thighs, arms and trunk . In the preceding 2 years he had developed new lesions on both hands . He had been previously diagnosed with sarcoidosis on the basis of skin and visceral histology, but subsequent opinion was that these were sarcoid-like granulomas rather than being representative of true sarcoidosis . Biopsy of the hand lesions showed necrotizing granulomas, and a single acid-fast bacillus (AFB) was identified on Wade-Fite stain . Subsequent repeat tissue biopsies for histology, culture and polymerase chain reaction testing failed to confirm the presence of mycobacterial organisms and it was felt that the organism was a contaminant introduced during tissue processing . The hand lesions responded well to intralesional injections of triamcinolone acetonide 10 mg/mL and oral tetracycline 500 mg b.d . was later introduced with a good clinical response . The diagnostic dilemma of finding granulomatous inflammation in a patient with common variable immunodeficiency, and the significance of a single AFB on histology are discussed . The treatment of sarcoid-like granulomas with tetracycline therapy is also commented on. Mar Biotechnol (NY), 2002 Mar, 4(2), 179 - 88 Characterization of surfactin-like cyclic depsipeptides synthesized by Bacillus pumilus from ascidian Halocynthia aurantium; Kalinovskaya NI et al.; A marine bacterium (KMM 1364), identified as Bacillus pumilus, was isolated from the surface of ascidian Halocynthia aurantium . Structural analysis revealed that the strain KMM 1364 produced a mixture of lipopeptide surfactin analogs with major components with molecular masses of 1035, 1049, 1063, and 1077 . The variation in molecular weight represents changes in the number of methylene groups in the lipid and/or peptide portions of the compounds . Structurally, these lipopeptides differ from surfactin in the substitution of the valine residue in position 4 by leucine, and have been isolated as two carboxy-terminal variants, with valine or isoleucine in position 7 . As constituents of the lipophilic part of the peptides, only beta-hydroxy-C(15)-, beta-hydroxy-C(16)-, and a high amount of beta-hydroxy-C(17) fatty acid were determined. Mar Biotechnol (NY), 2002 Mar, 4(2), 111 - 8 Antifouling potential of some marine organisms from India against species of Bacillus and Pseudomonas; Bhosale SH et al.; Crude methanolic extracts of 37 marine organisms (16 species of flora, 21 species of fauna) were screened for antibacterial properties against 5 strains of bacteria isolated from marine environments . Of these, 10 plant and 9 animal extracts exhibited antibacterial activity against at least one bacterial strain . The extracts of 6 species were active against all the strains: i.e., Stoechospermum marginatum (brown algae), Cymodocea rotundata (seagrass), Petrosia sp . and Psammaplysilla purpurea (sponges), Sinularia compressa (soft coral), and Cassiopeia sp . (jellyfish) . Among the plants, Padina tetrastromatica (brown algae) extract exhibited significant activity (9-11-mm inhibition zone at 500 microg per 6-mm disc) against Bacillus pumilus and Pseudomonas vesicularis, while the extracts of Petrosia, Psammaplysilla, and Cassiopeia were strongly active (11-13-mm inhibition zone at 500 microg per 6-mm disc) against B . circulans and P . putida . It was further confirmed that the attachment of bacterial strains on glass slides was inhibited remarkably with increasing concentrations of bioextracts of Petrosia sp . and Psammaplysilla purpurea . The present findings could form the basis for exploring the antibacterial potential of bioactive molecules from some of the marine organisms that exhibited moderate to strong antibacterial properties. Appl Microbiol Biotechnol, 2004 Aug, 65(2), 193 - 9 Epub 2004 Feb 11. Structural and functional characterization of the Bacillus megaterium uvrBA locus and generation of UV-sensitive mutants; Nahrstedt H et al.; The Bacillus megaterium genes uvrB and uvrA, encoding two subunits of the (A)BC excinuclease, which is responsible for nucleotide excision repair, were isolated and functionally characterized . RNA analyses revealed co-transcription of both genes probably forming a bicistronic operon . Expression of uvrB and uvrA was inducible by the DNA-damaging agent mitomycin C . This finding agrees with the presence of a potential DinR box within the uvrBA promoter . Single inactivation of uvrB or uvrA as well as the parallel knockout of both genes resulted in mutants highly sensitive to UV irradiation . Thus, this locus represents an attractive target for generating biologically safe containment strains of B . megaterium. Insect Biochem Mol Biol, 2004 Mar, 34(3), 193 - 202 Fluorescent-based assays establish Manduca sexta Bt-R(1a) cadherin as a receptor for multiple Bacillus thuringiensis Cry1A toxins in Drosophila S2 cells; Hua G et al.; A fluorescence-based approach was developed to analyze in vivo the function of Manduca sexta cadherin (Bt-R(1)) as a Cry1 toxin receptor . We cloned a Bt-R(1a) cDNA that differs from Bt-R(1) by 37 nucleotides and two amino acids and expressed it transiently in Drosophila melanogaster Schneider 2 (S2) cells . Cells expressing Bt-R(1a) bound Cry1Aa, Cry1Ab, and Cry1Ac toxins on ligand blots, and in saturation binding assays . More Cry1Ab was bound relative to Cry1Aa and Cry1Ac, though each Cry1A toxin bound with high-affinity (Kd values from 1.7 to 3.3 nM) . Using fluorescent microscopy and flow cytometry assays, we show that Cry1Aa, Cry1Ab and Cry1Ac, but not Cry1Ba, killed S2 cells expressing Bt-R(1a) cadherin . These results demonstrate that M . sexta cadherin Bt-R(1a) functions as a receptor for the Cry1A toxins in vivo and validates our cytotoxicity assay for future receptor studies. Oral Microbiol Immunol, 2004 Apr, 19(2), 88 - 94 The effect of oxygen on the growth and physiology of Porphyromonas gingivalis; Diaz PI et al.; Oxygen constitutes a constant challenge for the survival of strict anaerobes in the oral environment . The aim of this study was to investigate the effect of oxygen on the physiology and growth of Porphyromonas gingivalis in a continuous culture system when grown under conditions of hemin limitation and excess . Results showed that, when grown in the presence of hemin at 0.5 mg/l, P . gingivalis could tolerate low levels of oxygen, being able to reach steady-state when 6% oxygen was present in the incoming gas mixture . When the hemin concentration was increased to 5 mg/l, the culture tolerated 10% oxygen . Anaerobically-grown cells were coccoid in shape, whereas those grown in the presence of oxygen were bacillary . Acetate was the predominant end-product in cultures grown in the presence of oxygen or in cultures hemin-limited . Despite some changes in the activity of Arg- and Lys-gingipain, most of the proteolytic activity was retained in the presence of oxygen . Activity of each of the three anti-oxidant enzymes tested (NADH oxidase, NADH peroxidase and SOD) was detected under all conditions and usually increased under oxygenated environments . Higher activities were also seen in the hemin-limited cultures . These results show some of the changes that occur in the physiology of P . gingivalis as a result of oxidative stress and confirm that hemin has a protective effect on the growth of the microorganism in the presence of oxygen. Biotechnol Appl Biochem, 2004 Oct, 40(Pt 2), 191 - 6 Optimization of medium composition for keratinase production on feather by Bacillus licheniformis RG1 using statistical methods involving response surface methodology; Ramnani P et al.; A 3.5-fold increase in keratinase production by Bacillus licheniformis RG1 was achieved by using statistical methods involving Plackett-Burman design and response surface methodology . Eight variables were screened using Plackett-Burman design . Of these, glucose, peptone and glutathione