Int J Food Microbiol, 2001 Jan 22, 63(1-2), 35 - 50 Investigation of bacterial spore structure by high resolution solid-state nuclear magnetic resonance spectroscopy and transmission electron microscopy; Leuschner RG et al.; High resolution solid-state nuclear magnetic resonance spectroscopy (NMR) in combination with transmission electron microscopy (TEM) of spores of Bacillus cereus, an outer coatless mutant B . subtilis 322, an inner coatless mutant B . subtilis 325 and of germinated spores of B . subtilis CMCC 604 were carried out . Structural differences in the coats, mainly protein of spores were reflected by NMR spectra which indicated also differences in molecular mobility of carbohydrates which was partially attributed to the cortex . Dipicolinic acid (DPA) of spores of B . cereus displayed a high degree of solid state order and may be crystalline . Heat activation was studied on spores of B . subtilis 357 lux + and revealed a structural change when analysed by TEM but this was not associated with increases in molecular mobility since no effects were measured by NMR. Int J Food Microbiol, 2001 Jan 22, 63(1-2), 29 - 34 Modelling the influence of pH and organic acid types on thermal inactivation of Bacillus cereus spores; Leguerinel I et al.; A model is proposed to describe the influence pH on the heat resistance of Bacillus cereus spores . In addition to the conventional z value, the effect of pH on the thermal resistance of spores is characterised by a z(pH) value (z(pH) is the distance of pH from a reference pH*, which leads to a 10-fold reduction of D value) . The type of organic acid used for acidifying the heating medium, influences the z(pH) value . For nine organic acids, a linear relationship between the calculated z(pH) value and its lower acid pKa is observed . This relationship showed that the acid form (dissociated or undissociated) modifies the thermal spore resistance in addition to the H+ ion . The influence of acetic acid concentration on the D value at pH 7 shows the protective effect of the dissociated acid form on the heat resistance of spores . The acid concentration in the medium modified the heat resistance of spore and the z(pH) value. Arch Virol, 2000, 145(12), 2643 - 57 Genetic composition and complexity of virus populations at tungro-endemic and outbreak rice sites; Azzam O et al.; We have recently demonstrated the geographic isolation of rice tungro bacilliform virus (RTBV) populations in the tungro-endemic provinces of Isabela and North Cotabato, Philippines . In this study, we examined the genetic structure of the virus populations at the tungro-outbreak sites of Lanao del Norte, a province adjacent to North Cotabato . We also analyzed the virus populations at the tungro-endemic sites of Subang, Indonesia, and Dien Khanh, Vietnam . Total DNA extracts from 274 isolates were digested with EcoRV restriction enzyme and hybridized with a full-length probe of RTBV . In the total population, 22 EcoRV-restricted genome profiles (genotypes) were identified . Although overlapping genotypes could be observed, the outbreak sites of Lanao del Norte had a genotype combination distinct from that of Subang or Dien Khanh but a genotype combination similar to that identified earlier from North Cotabato, the adjacent endemic province . Sequence analysis of the intergenic region and part of the ORF1 RTBV genome from randomly selected genotypes confirms the geographic clustering of RTBV genotypes and, combined with restriction analysis, the results suggest a fragmented spatial distribution of RTBV local populations in the three countries . Because RTBV depends on rice tungro spherical virus (RTSV) for transmission, the population dynamics of both tungro viruses were then examined at the endemic and outbreak sites within the Philippines . The RTBV genotypes and the coat protein RTSV genotypes were used as indicators for virus diversity . A shift in population structure of both viruses was observed at the outbreak sites with a reduced RTBV but increased RTSV gene diversity. In Vivo, 2000 Nov-Dec, 14(6), 721 - 4 Oxidative stress of red blood cells during Bacillus Calmette-Guerin intravesical instillations; Mitropoulos D et al.; Although Bacillus Calmette-Guerin (BCG) intravesical instillation is widely accepted as a very effective modality in treating bladder carcinoma in situ, and in preventing superficial bladder cancer recurrence, its mechanism of action is not yet fully understood . The antitumor effects of BCG are mostly related to local immunological events but a systemic activation of the immune system cannot be excluded . The objective of the present study was to estimate the systemic production of oxidants during intravesical BCG treatment . Systemic production of oxidants was estimated by assessing the red blood cells (RBC) oxidative stress in twelve patients undergoing BCG immunotherapy for bladder carcinoma in situ . RBC oxidative stress induced by peroxynitrite was determined by luminol-enhanced chemiluminescence . During the treatment period, the RBC oxidative stress revealed a biphasic curve of changes: after an initial 5-fold increase, it dropped to pretreatment levels following the 4th instillation . Intravesical BCG administration induced systemic production of oxygen free radicals that may reflect a systemic activation of the immune system. Z Naturforsch {C}, 2000 Nov-Dec, 55(11-12), 987 - 90 PCR-based identification of Bacillus thuringiensis isolated from soil samples in Nigeria; Ogunjimi AA et al.; Six isolates of Bacillus thuringiensis isolated from soil samples confirmed to be toxic to mosquito larvae were differentiated using a PCR-Based technique . Three of these isolates initially identified using a serological technique were further differentiated with the PCR amplification of the delta-endotoxin target sequences . Using the total DNA of isolates as template, at least four isolates yielded amplicons one or all the crystal protein genes, cryI a, b, c, or II with sizes ranging from 238-1070 bp . None of these isolates yielded an amplicon for any of Cry IV A, B and D tested . Of the four isolates identified by PCR technique one isolate remained unidentified by serology. Lipids, 2000 Dec, 35(12), 1371 - 5 Unusual lipid composition of a Bacillus sp . isolated from Lake Pomorie in Bulgaria; Carballeira NM et al.; The lipid composition of a Bacillus sp., isolated from Lake Pomorie in Bulgaria, was unusual and consisted of 26 different fatty acids between C12 and C26, with anteiso C15-C17 saturated fatty acids predominating . The furan fatty acid, 10,13-epoxy-11-methyloctadeca-10,12-dienoic acid, was also identified, a new finding for this genus . The hydrocarbons consisted of 30 different monounsaturated hydrocarbons, between C25 and C30, with the iso-iso, iso-anteiso, anteiso-anteiso, iso-normal, and anteiso-normal methyl branching for odd-numbered chains, and the iso-iso, iso-anteiso, iso-normal, and anteiso-normal methyl branching for even-numbered chains . The double bond positions in these hydrocarbons were determined by dimethyl disulfide derivatization followed by GC-MS, and the double-bond cis configuration was confirmed by infrared spectroscopy . Some previously unknown hydrocarbons in bacteria, such as (Z)-3,21-dimethyl-9-tricosene, (Z)-3,21-dimethyl-10-tricosene, (Z)-2,24-dimethyl-11-pentacosene, and (Z)-2,25-dimethyl-13-hexacosene were identified . Sterols were detected and were based on the sitosterol nucleus. Nagoya J Med Sci, 2000 Nov, 63(3-4), 123 - 8 Severe disseminated BCG infection in an 8-year-old girl; Yamanaka K et al.; An 8-year-old girl died of sepsis due to staphylococcal infection one year and 8 months after Bacille Calmette-Guerin (BCG) revaccination . Two months after the vaccination in accordance with the school health program, she was hospitalized with a high fever, skin rash over the face and lower limbs, and leukopenia . Her clinical and laboratory pictures were not compatible with those of any established type of immunodeficiency . The polymerase chain reaction (PCR) test for M . tuberculosis complex was positive for bone marrow, pleural fluid, and peripheral blood . The strain recovered from a mycobacterial culture of the blood was identical to the BCG strains with which the patient was vaccinated, based on restriction fragment length polymorphism (RFLP) and a pulse-field gel electrophoresis (PFGE) analyses of DNA . She developed finally a lung abscess due to staphylococcal septicemia, which was the direct cause of her death. Exp Appl Acarol, 2000 Jul, 24(7), 497 - 560 Diseases of mites; van der Geest LP et al.; An overview is given of studies on diseases of mites . Knowledge of diseases of mites is still fragmentary but in recent years more attention has been paid to acaropathogens, often because of the economic importance of many mite species . Most research on mite pathogens concerns studies on fungal pathogens of eriophyoids and spider mites especially . These fungi often play an important role in the regulation of natural mite populations and are sometimes able to decimate populations of phytophagous mites . Studies are being conducted to develop some of these fungi as commercial acaricides . Virus diseases are known in only a few mites, namely, the citrus red mite and the European red mite . In both cases, non-occluded viruses play an important role in the regulation of mite populations in citrus and peach orchards, respectively, but application of these viruses as biological control agents does not seem feasible . A putative iridovirus has been observed in association with Varroa mites in moribund honeybee colonies . The virus is probably also pathogenic for honeybees and may be transmitted to them through this parasitic mite . Few bacteria have been reported as pathogens of the Acari but in recent years research has been concentrated on intracellular organisms such as Wolbachia that may cause distorted sex ratios in offspring and incompatibility between populations . The role of these organisms in natural populations of spider mites is in particular discussed . The effect of Bacillus thuringiensis on mites is also treated in this review, although its mode of action in arthropods is mainly due to the presence of toxins and it is, therefore, not considered to be a pathogen in the true sense of the word . Microsporidia have been observed in several mite species especially in oribatid mites, although other groups of mites may also be affected . In recent years, Microsporidia infections in Phytoseiidae have received considerable attention, as they are often found in mass rearings of beneficial arthropods . They affect the efficacy of these predators as biological control agent of insect and mite pests . Microsporidia do not seem to have potential for biological control of mites. J Mol Microbiol Biotechnol, 2001 Jan, 3(1), 73 - 82 Protein-protein interaction between Bacillus stearothermophilus tyrosyl-tRNA synthetase subdomains revealed by a bacterial two-hybrid system; Karimova G et al.; We have recently developed a bacterial two-hybrid system (BACTH), based on functional complementation between two fragments of the catalytic domain of Bordetella pertussis adenylate cyclase (AC), that allows an easy in vivo screening and selection of functional interactions between two proteins in Escherichia coli . In this work, we have further explored the potentialities of the BACTH system to study protein-protein interactions, using as a model, the interactions between various subdomains of the dimeric tyrosyl-tRNA synthetase (TyrRS) of Bacillus stearothermophilus . Using the BACTH system we confirmed the known interactions of the alpha/beta domains and those between the alpha/beta domain and the alpha domain that could be anticipated from the three-dimensional structure of TyrRS . Interestingly, the BACTH system revealed the unexpected interaction between the TyrRS alpha domains which is presumably mediated by a pseudo-leucine zipper motif . This study illustrates the interest of the bacterial two-hybrid system to delineate interacting domains of proteins and shows that it can reveal interactions that occur in vivo and that were not anticipated from the three-dimensional structure of the protein of interest. J Leukoc Biol, 2001 Jan, 69(1), 138 - 48 Impaired IL-15 production associated with susceptibility of murine AIDS to mycobacterial infection; Umemura M et al.; LP-BM5 murine leukemia virus (MuLV) injection causes murine AIDS (MAIDS), a disease characterized by many functional abnormalities of immunocompetent cells . We show that MAIDS mice are susceptible to Mycobacterium bovis Bacille Calmette-Guerin (BCG) infection as assessed by survival rate and bacterial counts . The peritoneal exudate macrophages from MAIDS mice produced a significant level of interleukin (IL)-12 soon after inoculation with BCG, whereas IL-15 and tumor necrosis factor (TNF) production were severely impaired in BCG-infected MAIDS mice . The appearance of natural killer (NK) and CD4+ T helper type 1 (Th1) cells specific for mycobacterial antigen were depressed in MAIDS mice after BCG infection . Thus, it appeared that impaired production of IL-15, besides other inflammatory cytokines, in MAIDS mice may be involved in the poor responses of the NK and Th1 cells, resulting in an increased susceptibility to BCG. J Basic Microbiol, 2000, 40(5-6), 385 - 8 Diversity of Bacillus thuringiensis in different habitats of northern Jordan; Obeidat M et al.; A survey of Bacillus thuringiensis was conducted for 17 locations in Northern Jordan representing 12 different habitats . Eighty isolates were identified as B . thuringiensis in the majority of the tested samples . Results showed that soils contaminated with the slaughterhouses waste materials had the highest content of spore-forming bacteria {(4.05-2.2) 10(7) CFU/g)} and B . thuringiensis {(4.05-7.9) 10(7) CFU/g)} with a (5.5%-14.9%) and (5.2%-7.7%) of the total viable bacterial count, respectively . These bacteria were more abundant in soils contaminated with such animal by-products. J Basic Microbiol, 2000, 40(5-6), 363 - 8 New enzymatic method for determining D-arabinitol in serum; Hino M et al.; A new reagent has been developed to determine D-arabinitol . This utilizes D-arabinitol 2-oxidoreductase derived from Bacillus sp . with high stability, and water-soluble tetrazolium salt, that can detect NADH with high sensitivity . Since this enzyme does not react to D-mannitol, elimination of D-mannitol is unnecessary . Thus, this is a much simpler process than currently available with commercial kits use D-arabinitol 4-oxidoreductase . The within-run and between-run precisions (CV) were 2.4-6.9% and 3.1-8.7%, respectively, whilst the correlation (r) between the results obtained with our proposed method (y) and those obtained with the commercial "Arabinitec-auto" kit (x) was 0.964 (y = 1.02x + 0.933 mumol/l; n = 69) . However, some samples deviated remarkably from correlation in both methods . Our analyzing accuracy is satisfactory in clinical application, as it does not miss positive sample over cut-off value . We are refining this method by investigating why some specimens are apart from correlation significantly. J Am Mosq Control Assoc, 2000 Dec, 16(4), 342 - 5 Evaluation of methylated soy oil and water-based formulations of Bacillus thuringiensis var . Israelensis and Golden Bear Oil (GB-1111) against anopheles quadrimaculatus larvae in small rice plots; Dennett JA et al.; The efficacy of formulations containing methylated soybean oil (MSO) alone and with technical-grade Bacillus thuringiensis var . israelensis (Bti) were compared to Golden Bear Oil (GB-1111) and a water-based Bti formulation against 3rd- to 4th-stage Anopheles quadrimaculatus larvae confined to sentinel cages in small rice plots . Three replicates each of MSO with 2% Pyroter added as a surfactant (MSO + PYR), MSO with 2% Pyroter and 4 g of Bti technical powder (MSO + PYR + Bti), GB-1111, a water-based formulation with 4 g of Bti technical powder (Bti + water), and untreated controls were performed . Mosquito larvae were introduced on the 1st day of treatment and at 4 days posttreatment . Mortality was recorded at 24 and 48 h posttreatment for the 1st installation and at 5 days posttreatment for the 2nd installation . The Bti + water formulation provided 71% control and the MSO + PYR + Bti formulation achieved 64% control, whereas MSO + PYR and GB-1111 produced 16 and 18% control, respectively, at 24 h posttreatment . With the exception of MSO + PYR + Bti, which decreased by 2%, the mean percent control increased slightly at 48h posttreatment across remaining treatments, with Bti + water obtaining 72% control . This was significantly higher than GB-1111, which achieved 23% control at 48 h posttreatment . The MSO + PYR and MSO + PYR + Bti formulations yielded 56 and 62% control, respectively, during the same interval and were not significantly different from one another . Formulations containing MSO + PYR exhibited delayed activity similar to GB-1111, with all formulations except MSO + PYR + Bti providing greatest control at 48 h posttreatment . Both MSO formulations (MSO + PYR + Bti and MSO + PYR) were statistically comparable to Bti + water and GB-1111, respectively, at 24 and 48 h posttreatment . None of the formulations exhibited a residual activity adequate enough to control An . quadrimaculatus larvae for up to 5 days. J Am Mosq Control Assoc, 2000 Dec, 16(4), 321 - 3 Control of aedes aegypti breeding in desert coolers and tires by use of Bacillus thuringiensis var . Israelensis formulation; Batra CP et al.; Three different formulations of Bacillus thuringiensis var . israelensis (Bti) were evaluated for their efficacy against immature Aedes aegypti in desert coolers and tires . Three formulations, viz., VectoBac tablets, VectoBac granules, and Bacticide powder, at the application rate of 0.75, 2, and 1 g per cooler, respectively, and VectoBac tablets at 0.75 and 0.375 g per tire, were evaluated . In coolers and tires, 100% reduction in the abundance of late larval instars of Ae . aegypti was observed for a period of 2 and 3 wk, respectively . The possibility of using tablets and capsules filled with Bti granules and powder formulation by individuals or communities for control of Ae . aegypti breeding has been discussed in view of the increasing outbreaks of dengue and dengue hemorrhagic fever in India . Use of these formulations over conventional methods is better and more user-friendly. J Egypt Soc Parasitol, 2000 Dec, 30(3), 839 - 49 Potential for Culex pipiens to develop resistance against Bacillus sphaericus toxins; Soliman BA et al.; Susceptibility of field Culex pipiens populations to Bacillus sphaericus and the possibility of resistance development in the selected laboratory colony were investigated . 180 populations collected from Giza and Qualyobia over one year show no marked variation at LC50 and LC90 values . Larvae of laboratory colony of Cx pipiens were subjected to selection pressure with a preparation of Bacillus sphaericus strain 2362 at LC80 values for 20 successive generations . The selected laboratory colony began developing resistance at F10 (RR = 1.69) when compared with its unselected counterpart . The magnitude of resistance increased gradually and reached 33 fold by F20 . The unselected colony reared without any exposure to the B . sphaericus toxin, did not show any marked change in their susceptibility to this microbial agent. Faraday Discuss, 2000, (116), 135 - 53; discussion 171-90 Engineering artificial redox chains by molecular 'Lego'; Sadeghi SJ et al.; This work reports on a novel approach for building artificial redox chains: the molecular 'Lego' approach . This exploits the scaffold of natural redox proteins by fusing together functional protein modules with the desired properties . The molecular 'Lego' mimics the natural molecular evolution that proceeded by modular assembly of genes/DNA segments . Non-physiological electron transfer partners, flavodoxin (fld) and cytochrome c553 (c553) from Desulfovibrio vulgaris and the haem domain of P450 BM3 (BMP) from Bacillus megaterium have been used as building blocks in different combinations to build artificial redox chains . The kinetic characterization of the electron transfer (ET) between the separate building blocks has been carried out . Under pseudo-first order conditions, a limiting ET rate, klim, of 0.48 +/- 0.05 s-1 and 43.77 +/- 2.18 s-1 and an apparent binding constant, Kapp, of 21 +/- 6 microM and 1.23 +/- 0.32 microM have been found for the fld/c553 and fld/BMP redox pairs, respectively . These results show that fld can be used as a module for transferring electrons to c553 and BMP . A 3D model of the fld/c553 and fld/BMP complexes was used to guide the construction of covalently linked assemblies via engineered disulfide bridges or by fusion of the relevant genes via an engineered loop . The first approach led to the construction, expression and characterization of the S35C and S64C mutants of fld and M23C and G51C mutants of c553 . Although the redox potentials of the separate mutants were found to be the same as those of recombinant wild type proteins (-408 mV for the semiquinone/hydroquinone couple of fld and +32 mV for the c553), the c553 homo-dimers M23C-M23C and G51C-G51C were found to have redox potentials of +88 and +105 mV, respectively . These differences have been analysed in terms of exposure of the haem cofactors to the solvent, and these lead to some interesting questions on the redox potentials of the transient redox complexes in physiological systems . The fld-c553 S64C-M23C and S35C-M23C chimeras were constructed, expressed and purified but the FMN was found to be destabilised resulting in the apo-form of these proteins . The gene fusion strategy was used to produce covalently linked assemblies of both fld-c553 and fld-BMP . The former was expressed using a seven amino acid (GPGPGPG) loop linking the C-terminus of fld to the N-terminus of c553 . The fld-BMP fusion protein was successfully expressed by using the naturally occurring loop of the P450 BM3 (residues 471-479) to link the BMP domain at the N-terminus with fld domain at the C-terminus . This fusion was found to be correctly folded and functional . Efficient ET from the FMN to the haem domain (370 s-1) was also found to be in the same region of the physiological redox partners (250 s-1) . This work demonstrates the feasibility of the molecular 'Lego' approach in generating functional multi-domain proteins with designed properties, beyond the restrictions imposed by the naturally occurring protein domains. Arch Tierernahr, 2000, 53(4), 353 - 73 Comparative studies on the in vitro properties of phytases from various microbial origins; Igbasan FA et al.; The physical and chemical properties of six crude phytase preparations were compared . Four of these enzymes (Aspergillus A, Aspergillus R, Peniophora and Aspergillus T) were produced at commercial scale for the use as feed additives while the other two (E . coli and Bacillus) were produced at laboratory scale . The encoding genes of the enzymes were from different microbial origins (4 of fungal origin and 2 of bacterial origin, i.e., E . coli and Bacillus phytases) . One of the fungal phytases (Aspergillus R) was expressed in transgenic rape . The enzymes were studied for their pH behaviour, temperature optimum and stability and resistance to protease inactivation . The phytases were found to exhibit different properties depending on source of the phytase gene and the production organism . The pH profiles of the enzymes showed that the fungal phytases had their pH optima ranging from 4.5 to 5.5 . The bacterial E . coli phytase had also its pH optimum in the acidic range at pH 4.5 while the pH optimum for the Bacillus enzyme was identified at pH 7.0 . Temperature optima were at 50 and 60 degrees C for the fungal and bacterial phytases, respectively . The Bacillus phytase was more thermostable in aqueous solutions than all other enzymes . In pelleting experiments performed at 60, 70 and 80 degrees C in the conditioner, Aspergillus A, Peniophora (measurement at pH 5.5) and E . coli phytases were more heat stable compared to other enzymes (Bacillus enzyme was not included) . At a temperature of 70 degrees C in the conditioner, these enzymes maintained a residual activity of approximately 70% after pelleting compared to approximately 30% determined for the other enzymes . Incubation of enzyme preparations with porcine proteases revealed that only E . coli phytase was insensitive against pepsin and pancreatin . Incubation of the enzymes in digesta supernatants from various segments of the digestive tract of hens revealed that digesta from stomach inactivated the enzymes most efficiently except E . coli phytase which had a residual activity of 93% after 60 min incubation at 40 degrees C . It can be concluded that phytases of various microbial origins behave differently with respect to their in vitro properties which could be of importance for future developments of phytase preparations . Especially bacterial phytases contain properties like high temperature stability (Bacillus phytase) and high proteolytic stability (E . coli phytase) which make them favourable for future applications as feed additives. Mikrobiologiia, 2000 Nov-Dec, 69(6), 796 - 800 {Characterization of a new strain of Bacillus thuringiensis, a producer of an endotoxin against coleoptera}; Dobrzhanskaia EO et al.; A new strain of Bacillus thuringiensis 2-7 was found to belong to the serotype H8 . Cells of this strain contained irregular and flat crystalline inclusions and two large plasmids . The gene responsible for crystal formation is most likely located on the large plasmid greater than 105 MDa in size . Comparison of the cry gene of B . thuringiensis 2-7 and the cryIIIA gene of B . thuringiensis subsp . tenebrionis showed that their nucleotide sequences are identical. Acta Paediatr, 2000 Dec, 89(12), 1495 - 7 Sternal tuberculosis in a 9-month-old infant after BCG vaccination; Kato Y et al.; Tuberculosis osteitis is mainly observed as a late complication of the pulmonary infection . We describe a 9-mo-old Japanese infant who became infected with Mycobacterium tuberculosis even after receiving Bacillus Calmette-Guerin (BCG) vaccination at 3 mo of age . This is the first report that the sternum is the localization for tuberculous osteitis in infants . He developed a localized tumor in the sternum without any respiratory symptom . Mycobacterium tuberculosis complex was detected by means of nucleic acid amplification test . Gastric aspirates also yielded Mycobacterium tuberculosis . The source of the infection was unclear . CONCLUSION: Tuberculous osteitis should be excluded in infants with undiagnosed bone lesions, even if they have been vaccinated with BCG. Sci Total Environ, 2000 Dec 18, 263(1-3), 155 - 60 The measurement of volatile constituents in Foray 48B, an insecticide prepared from Bacillus thuringiensis var . kurstaki; van Netten C et al.; Foray 48B, an insecticide prepared from Bacillus thuringiensis var . kurstaki (Btk), has been used for many years to combat infestations of Gypsy moths . Foray 48B also contains a large number of 'inert ingredients' which are not disclosed by the manufacturer . Gypsy moths usually enter the country through marine- and airports in close proximity to urban areas, which consequently need to be sprayed . The population affected often demands more detailed information than what is available including the potential presence of volatile organic agents which could be released during spraying, posing a potential health hazard . Four different methods were investigated using GC/mass spectrometry regarding their ability to capture volatile agents associated with Foray 48B . It was found that solid phase micro-extraction was most efficient in capturing volatile agents from the head-space of Foray 48B . Separate trials using 95:5% ethanol/isopropanol mixture and toluene in an impinger configuration were much less efficient . Standard techniques using activated charcoal tubes in the laboratory setting as well as in a field trial did not capture any compounds . It was concluded that the volatile agents associated with Foray 48B did not appear to constitute a significant health hazard and no one agent was a likely candidate to serve as a tracer for Foray 48B exposure. AIDS, 2001 Jan 5, 15(1), 55 - 60 Bacteremia due to Mycobacterium tuberculosis or M . bovis, Bacille Calmette-Guérin (BCG) among HIV- positive children and adults in Zambia; Waddell RD et al.; BACKGROUND: Among adults with advanced HIV infection in developing countries, bacteremia due to Mycobacterium tuberculosis (MTB) is common and bacteremia due to M . bovis (bacille Calmette-Guerin; BCG) is rare . Comparable data are not available for children with HIV . OBJECTIVE: To compare the prevalence of bacteremia due to M . tuberculosis or M . bovis BCG in hospitalized children and adults with HIV infection in a developing country with a high prevalence of tuberculosis and HIV and > 95% BCG immunization coverage . DESIGN: Descriptive cross-sectional study . METHODS: Prospectively hospitalized patients in Lusaka, Zambia who were suspected to have HIV infection underwent phlebotomy for HIV ELISA, HIV viral load, and lysis-centrifugation blood culture for mycobacteria . Histories were obtained and patients were examined for BCG scars . Mycobacterial isolates were identified using DNA probes for MTB complex (MTBC), multiplex PCR and IS6110 typing . RESULTS: The median age of 387 HIV-positive children was 15 months; 98% were BCG immunized . The median age of 344 HIV-positive adults was 32 years; 44% were BCG immunized . Blood cultures were positive for mycobacteria in six children (2%) and 38 adults(11%) (P < 0.001) . The six pediatric isolates included five MTBC (40% clustered) and one BCG . The 38 adult isolates included 36 MTBC (16% clustered) and two M . avium complex . CONCLUSION: Bacteremia due to MTB is less common among children than adults with advanced HIV infection in Zambia . Bacteremia due to M . bovis BCG is rare even among children with recent BCG immunization and symptomatic HIV infection. J Med Microbiol, 2001 Jan, 50(1), 23 - 8 Binding of alpha2-laminins by pathogenic and non-pathogenic mycobacteria and adherence to Schwann cells; Marques MA et al.; The ability of Mycobacterium leprae to specifically bind alpha2-laminins of Schwann cells has been described recently as being an important property of the leprosy bacillus, which could explain the neural tropism of M . leprae . Therefore, the extent of the expression of alpha2-laminin-binding properties among mycobacteria was investigated . In an ELISA-based assay, all three species of Mycobacterium tested (M . tuberculosis, M . chelonae and M . smegmatis) expressed laminin-binding capacity, suggesting that the ability to bind alpha2-laminins is conserved within the genus Mycobacterium . This report also demonstrated that not only M . leprae but all the mycobacterial species tested readily interacted with the ST88-14 cells, a human schwannoma cell line, and that the addition of soluble alpha2-laminins significantly increased their adherence to these cells . These results failed to demonstrate the presence in M . leprae of a unique system based on alpha2-laminins for adherence to Schwann cells. Yi Chuan Xue Bao, 2000, 27(10), 932 - 8 {Cloning of plasmid pBMB2062 in Bacillus thuringiensis strain YBT-1520 and construction of plasmid vector with genetic stability}; Sun M et al.; A small plasmid pBMB2062 in Bacillus thuringiensis strain YBT-1520 was cloned and sequenced . Its 2,062 bp sequence contains two potential open reading frames (orfs) . The orf1 and orf2 encode a tentative replication initial protein consisting of 289 amino acid residues and a tentative replication protein consisting of 80 amino acid residues, respectively . Two homological plasmids were found by Blast searching . There are 23 nucleotides difference occurring among three of the plasmids . The difference occurred in the orf1 causes different encoding capability . Comparing with the orf1 in pBMB2062, the orf1 in the homological plasmids are truncated, one at the N-terminal and another at the C-terminal . cDNA synthesis and PCR detection showed that the mRNA corresponding to orf1 in pBMB2062 really occurs . Shuttle vectors were constructed based on pBMB2062 and showed the ability to express insecticidal crystal gene . Under nonselective condition, recombinant plasmids based on pBMB2062 were genetically stable. Sheng Wu Gong Cheng Xue Bao, 2000 Sep, 16(5), 587 - 90 {Transfer of cry1C gene into Bacillus thuringiensis by electroporation to construct strain with broader insecticidal activity}; Lu SQ et al.; Three Transformants were selected by transferring cry1C into Bacillus thuringiensis strain YBT1535 . Plasmid profiles, PCR and Southern blot result all proved that cry1C had been transferred into strain YBT1535 . Bioassay results showed that the transformants of strain YBT1535 displayed significantly higher toxicity against Spodoptera exigua than strain YBT1535, but the toxicities against Heliothis armigera and Plutella xylostella did not rise except transformant YBT1535-2. Med Clin North Am, 2001 Jan, 85(1), 79 - 114 Nosocomial pneumonia . Diagnostic and therapeutic considerations; Cunha BA; Many patients with presumed nosocomial pneumonia probably have infiltrates on the chest radiograph, fever, and leukocytosis resulting from noninfectious causes . Because of the high mortality and morbidity associated with nosocomial pneumonias, however, most clinicians treat such patients with a 2-week empiric trial of antibiotics . Before therapy is initiated, the clinician should rule out other causes of pulmonary infiltrates, fever, and leukocytosis that mimic a nosocomial pneumonia (e.g., pre-existing interstitial lung disease, primary or metastatic lung carcinomas, pulmonary emboli, pulmonary drug reactions, pulmonary hemorrhage, collagen vascular disease affecting the lungs, or congestive heart failure) . If these disorders can be eliminated from diagnostic consideration, a 2-week trial of empiric monotherapy is indicated . The clinician should treat cases of presumed nosocomial pneumonia as if P . aeruginosa were the pathogen . Although P . aeruginosa is not the most common cause of nosocomial pneumonia, it is the most virulent pulmonary pathogen associated with nosocomial pneumonia . Coverage directed against P . aeruginosa is effective against all other aerobic gram-negative bacillary pathogens causing hospital-acquired pneumonia . The clinician should select an antibiotic for empiric monotherapy that is highly effective against P . aeruginosa, has a good side-effect profile, has a low resistance potential, and is relatively inexpensive in terms of its cost to the institution . The preferred agents for empiric monotherapy for nosocomial pneumonia are cefepime, meropenem, and piperacillin . Single organisms are responsible for nosocomial pneumonia, not multiple pathogens . S . aureus rarely, if ever, causes nosocomial pneumonia but is mentioned frequently in studies based on cultures of respiratory tract secretions . S . aureus, unless accompanied by a necrotizing pneumonia with rapid cavitation within 72 hours, in the sputum indicates colonization rather than infection and should not be addressed therapeutically . Antibiotics associated with a high resistance potential should not be used as monotherapy or included in combination therapy regimens (i.e., ceftazidime, ciprofloxacin, imipenem, or gentamicin) . Combination therapy is more expensive than monotherapy and is indicated only when P . aeruginosa is extremely likely, based on its characteristic clinical presentation, or is proved by tissue biopsy . Therapy should not be based on respiratory secretion cultures regardless of technique . Optimal combination regimens include cefepime or meropenem plus levofloxacin or piperacillin or aztreonam or amikacin . Nosocomial pneumonias usually are treated for 14 days . Lack of radiographic or clinical response to appropriate empiric nosocomial pneumonia monotherapy after 14 days suggests an alternate diagnosis . In these patients, a tissue biopsy specimen should be obtained to determine the cause of the persistence of pulmonary infiltrates unresponsive to appropriate antimicrobial therapy. Rev Sci Tech, 2000 Apr, 19(1), 136 - 50 Cat-scratch disease; Chomel BB; Cat-scratch disease (CSD) was first described by Debre in 1950, yet the causative bacterial agent of CSD remained obscure until 1992, when Bartonella (formerly Rochalimaea) henselae was implicated in CSD by serological and microbiologic studies . Bartonella henselae had initially been linked to bacillary angiomatosis (BA), a vascular proliferative disease most commonly associated with long-standing human immunodeficiency virus infection or other significant immunosuppression . Bartonella henselae has also been associated with bacillary peliosis, relapsing bacteraemia and endocarditis in humans . Cats are healthy carriers of B . henselae, and can be bacteraemic for months or years . Cat-to-cat transmission of the organism by the cat flea, with no direct contact transmission, has been demonstrated . Two new Bartonella species have been identified in the cat reservoir, namely: B . clarridgeiae and B . koehlerae . The role of these species in the aetiology of CSD still needs to be confirmed by isolation or DNA identification from lesions in humans . The author discusses the present state of knowledge on the aetiology, clinical features and epidemiological characteristics of CSD/BA, in addition to diagnosis, treatment and prevention. Wei Sheng Wu Xue Bao, 1997 Dec, 37(6), 480 - 2 {The preparing of Bacillus penetrans preparation and their controlling to root-knot nematodes}; Pan C et al.; In order to make use of B . penetrans the preparation of these bacteria was prepared successfully . Each gram of the specimen contains more than 3.1 x 10(8) spores according to determination by bloodcount plate . The method of its production are: First, infect the larvae of root-knot nematodes with the spores of B . penetrans . Second, inoculate infectable plants with the infected larvae . Third, beat the root-systems of inoculated plants after adding some water . The flowed liquid from a bronze sieve was mixed with the clear water, filtered, precipitated, air-dried under 5-10 degrees C, and grinded into fine powder, When these powder was added into the water which contained the larvae of root-knot nematodes, the spores of B . penetrans adhered to the cuticles of these larvae in 24 h . The pot experiments of tomato, chilli and cockscomb showed that the galls, egg-sacs, females, males and all-stages of larvae produced by the plant inoculated with the larvae infected with B . penetrans are about 75%-80% less than that without these bacteria . Thus it can be seen that the spores of B . penetrans can depress the populations of root-knot nematodes obviously . Therefore, it is a promising preparation for biological control. Wei Sheng Wu Xue Bao, 1997 Dec, 37(6), 473 - 6 {Fermentation conditions of engineering strain utilizing starch for production of alkaline proteinase}; Feng Y et al.; The highest activity of alkaline proteinase produced by Bacillus pumilus c172(pBX 96) transformant was 9,000 U/ml in shaking flask where corn meal and bean cake meal were carbon and nitrogen sources, respectively . The enzyme activity was raised under the conditions of initial pH 7.0, MgCl2 instead of MgSO4, and glucose (0.1%) in the substrate . The fermentation of c172(pBx 96) transformant was carried out with parameters of pH, reducing sugar, total sugar and enzyme activity. Wei Sheng Wu Xue Bao, 1997 Oct, 37(5), 397 - 400 {The properties of protease from Bacillus sphaericus C3-41}; Sun F et al.; The production and properties of protease from Bacillus sphaericus strain C3-41 were reported in this paper . It was found out that the secrete of extracellular protease began at the exponential phase, reached a maximum at the early phase of sporangium and then decrease rapidly . The production conditions of the protease have been studied . The protease preparation was purified by salting out with ammonium sulfate and by chromatography fractionating on Sephadex G-100 . The purified enzyme have a specific activity of 6741.5 U/mg protein and a molecular weight of 42,000 . The optimal activities of the protease were around pH11.0 and at 4 degrees C respectively . The enzyme was stable at pH5.0-12.0 . The proteolytic activity was inhibited by phenylmethylsulphony fluoride (PMSF) and EDTA, but not by IAA, SA and DTT . The activity was inhibited in the presence of Al3+, Hg2+, Fe3+, Cu2+ and Fe2+ . The enzyme was sensitive to higher temperature, but was quite stable in the presence of Ca2+. Med Sci Monit, 2000 Jan-Feb, 6(1), 8 - 12 Factors influencing the cellular location of proteolytic enzymes of Bacillus intermedius; Sharipova MR et al.; Thiol-dependent serine proteinase and glutamylendopeptidase of Bacillus intermedius 3-19 being prevailing enzymes in the total pool of extracellular proteinases (95%) of this microorganism in catalytic active form were detected on the membrane of the cells . Production of these enzymes was maximum on the medium containing inorganic phosphate and gelatin and decreased 2-4-fold on the medium with glucose and lactate . The level of the activity of extracellular enzymes correlated with that of corresponding membrane-bound proteins . The addition of CoCl2 (2 mM) into the medium caused essential increase in extracellular glutamylendopeptidase activity and promoted the release of membrane-bound enzyme into cultural fluid . Proteolytic activity was detected in cytoplasm also . Proteinases localized in cytoplasm were shown to differ in properties from those secreted. Environ Microbiol, 1999 Dec, 1(6), 503 - 15 Climatic influence on mesophilic Bacillus cereus and psychrotolerant Bacillus weihenstephanensis populations in tropical, temperate and alpine soil; von Stetten F et al.; Bacillus weihenstephanensis strains are psychrotolerant and grow from below 7 degrees C to 38 degrees C . Closely related mesophilic Bacillus cereus strains can grow from above 7 degrees C to 46 degrees C . We classified 1060 B . cereus group isolates from different soil samples with respect to their psychrotolerant and mesophilic genotypes by polymerase chain reaction (PCR) targeting of specific 16S rDNA and cold shock protein A gene signatures . In parallel, growth tests at 7 degrees C were carried out to determine the thermal phenotype . The geographic distribution of psychrotolerant and mesophilic isolates was found to depend significantly on the prevalent annual average temperature . In one tropical, one temperate and two alpine habitats, the proportion of psychrotolerant cspA genotypes was found to be 0%, 45% and 86% and 98%, respectively, with the corresponding annual average temperatures being 28 degrees C, 7 degrees C, 4 degrees C and 1 degrees C . In the tropical habitat, only the mesophilic B . cereus was found, characterized by correspondence of thermal genotype and phenotype . In the alpine habitat, almost only the psychrotolerant B . weihenstephanensis was isolated . In the temperate habitat, mesophilic B . cereus and psychrotolerant B . weihenstephanensis as well as 'intermediate thermal types' occurred, the latter having opposite thermal genotypes and phenotypes or opposing sets of thermal DNA signatures, characterized by the coexistence of mesophilic and psychrotolerant 16S rDNA operon copies within a single isolate . Both sugar utilization and DNA fingerprinting patterns revealed a high, probably non-clonal microsite diversity within the population of the temperate habitat . We interpret our observations in terms of a temperature-dependent selection regime, acting on recombining B . cereus/ B . weihenstephanensis populations in soil. Clin Exp Immunol, 2001 Feb, 123(2), 264 - 70 Recombinant bacille Calmette-Guérin (BCG) expressing human interferon-alpha 2B demonstrates enhanced immunogenicity; Luo Y et al.; To increase its immunostimulatory properties, BCG was genetically engineered to secrete recombinant human interferon-alpha 2B (rhIFN-alpha) under control of the mycobacterial heat shock protein (hsp)60 promoter and the alpha antigen signal sequence . Expression of rhIFN-alpha was readily detectable by ELISA and on Western blotting . When compared with control BCG, rhIFN-alpha BCG was substantially more active in inducing the production of IFN-gamma and IFN-inducible protein 10 (IP-10) from human peripheral blood mononuclear cells, while IL-10 production was correspondingly decreased . These effects were reversible upon antibody neutralization of rhIFN-alpha . Among 10 patients tested, rhIFN-alpha BCG enhanced IFN-gamma production in all patients ranging from 1.4- to 23.7-fold with a general trend toward greatest enhancement among those with weakest baseline responses to control BCG . Correspondingly, rhIFN-alpha BCG decreased IL-10 production in all patients by 1.2-4.8-fold . The onset of IFN-gamma production induced by rhIFN-alpha BCG was also more rapid, occurring within 4 h after stimulation versus > 24 h with wild-type BCG . The observation that the maximum IFN-gamma induction depends on the simultaneous presence of both IFN-alpha and BCG highlights the advantages of rhIFN-alpha BCG . Taken together, these immunostimulatory properties of rhIFN-alpha BCG suggest that it may be a superior agent for immunotherapeutic protocols involving live BCG in humans. Clin Exp Immunol, 2001 Feb, 123(2), 219 - 25 Effect of deworming on human T cell responses to mycobacterial antigens in helminth-exposed individuals before and after bacille Calmette-Guérin (BCG) vaccination; Elias D et al.; The protective efficacy of BCG vaccination against pulmonary tuberculosis (TB) is highly variable in different populations . The reason remains to be elucidated . This study aims to investigate the possible effect of intestinal helminths on the immune response to PPD in naturally immunized or BCG-vaccinated humans . The study population was assessed for helminthic infection and those found to be positive were randomly assigned to either an albendazole treatment group or a control group who received a placebo . The immune response to PPD was compared between the two groups . In addition, subjects who were tuberculin skin test-negative in both groups were BCG vaccinated and later on tested for PPD-specific responses . Albendazole induced elimination/or reduction in intestinal worms resulting in a significant improvement in T cell proliferation and in interferon-gamma production by peripheral blood mononuclear cells (PBMC) stimulated with PPD . Moreover, BCG vaccination significantly improved PPD-specific immune responses in the treated group but not in the placebo group . The differences in the in vivo skin test responses were not significant . The data show that cellular immune responses to PPD are reduced in persons with concurrent helminthic infections, perhaps reflecting a lowered resistance to mycobacterial infections . This could explain, at least in part, the reduced efficacy of BCG against TB in helminth-endemic areas of the world. Cell Microbiol, 2000 Oct, 2(5), 431 - 41 Interaction of Bartonella henselae with endothelial cells results in rapid bacterial rRNA synthesis and replication; Kempf VA et al.; Bartonella henselae is a slow-growing microorganism and the causative pathogen of bacillary angiomatosis in man . Here, we analysed how interaction of B . henselae with endothelial cells might affect bacterial growth . For this purpose, bacterial rRNA production and ribosome content was determined by fluorescence in situ hybridization (FISH) using rRNA-targeted fluorescence-labelled oligonucleotide probes . B . henselae grown on agar plates showed no detectable rRNA content by means of FISH, whereas B . henselae co-cultured with endothelial cells showed a rapid increase of rRNA production within the first 18 h after inoculation . The increased rRNA synthesis was paralleled by a approximately 1000-fold intracellular bacterial replication, whereas bacteria grown on agar base showed only a approximately 10-fold replication within the first 48 h of culture . Pretreatment of host cells with paraformaldehyde prevented adhesion, invasion, intracellular replication and bacterial rRNA synthesis of B . henselae . In contrast, inhibition of host cell protein synthesis by cycloheximide did not affect bacterial adhesion and invasion, but prevented intracellular replication although bacterial rRNA content was increased . Inhibition of actin polymerization by cytochalasin D did not affect adhesion, invasion, increased rRNA content or intracellular replication of B . henselae . These results demonstrate that rRNA synthesis and replication of B . henselae is promoted by viable host cells with intact de novo protein synthesis. Oncol Rep, 2001 Mar-Apr, 8(2), 257 - 61 Changes in mucosal immune cells of bladder tumor patient after BCG intravesical immunotherapy; Chang SG et al.; The Bacillus Calmette-Guerin (BCG) is considered to be at least as effective, and perhaps superior to chemotherapy in the prophylaxis of recurrent superficial tumors . However, the mechanism of the antitumor effect of BCG is still not exactly known . We have conducted investigations to examine changes in bladder mucosal immune cells in patients with superficial bladder carcinoma treated with a first cycle of BCG . The study group included 15 BCG and 5 doxorubicin instillation patients, most in the intermediate or high risk group for recurrent tumor . Grossly normal bladder mucosal cold cup biopsies were performed at initial TUR and one week after six consecutive weekly instillations of BCG or doxorubicin . All specimens underwent immunohistochemical staining, both pre-treatment and post-treatment, including CD20, CD45RO, CD8, CD4 and CD57 . Immunoreactive cell counts were evaluated from three different microscopic fields (x400) under the grid . The mean duration of follow-up was 52.8 months . The post-treatment bladder mucosal B-cells (CD20) and T-cells (CD45RO, CD4, CD8) were significantly increased compared to pre-treatment in patients treated with BCG instillation, but NK-cells (CD57) were not changed . However, there was no change in B-cells or T-cells in patient treated with doxorubicin . The CD20 cells in pre-treatment specimens did not correlate with any other cells . However, it was a statistically significant correlation with CD45RO in post-treatment specimens . The CD4 correlated with CD45RO and CD8 in pre-treatment, but it was correlated with CD45RO and CD57 in post-treatment specimens . There was no tumor recurrence in cases with significantly increased B-cells after BCG instillation . The results of these studies suggest that intravesical BCG immunotherapy for superficial bladder tumor induces a significant increase in T-cells as well as B-cells and that B-cells have a preventive effect on tumor recurrence . Further studies with a larger number of patients are needed to confirm the value of the B-cell increment after BCG instillation as a clinically independent prognostic factor. Biotechnol Bioeng, 2001 Feb 20, 72(4), 434 - 40 Orthophosphate anion enhances the stability and activity of endoxylanase from Bacillus sp; Park MT et al.; Endoxylanase, for which the optimum temperature is 60 degrees C (optimum pH 7), is labile to heat . Because the isoelectric point (pI) value of this xylanase is 10.6, the net charge of this enzyme is positive at pH 7 . Thus, ions are likely to influence its enzyme structure and the thermal stability of endoxylanase may improve . Among the various ions tested, orthophosphate anion (HPO(4)(2-)) was found to significantly improve not only the stability but the activity of xylanase . When K(2)HPO(4) concentration was increased from 50 mM to 1.2 M, the T(m )value of xylanase was increased from 60.0 degrees C to 74.5 degrees C . The affinity of xylanase on xylan also increased along with K(2)HPO(4) concentration . Thus, the xylanase activity at 0.6 M K(2)HPO(4) was 2.3-fold higher than that at 50 mM K(2)HPO(4), and 120.2-fold higher than that in 40 mM MOPS buffer . This enhanced activity in the presence of K(2)HPO(4 )probably takes place because the orthophosphate anion affects the binding and catalytic residues of endoxylanase . Eur J Biochem, 2001 Feb, 268(4), 895 - 902 Properties of the prophenoloxidase activating enzyme of the freshwater crayfish, Pacifastacus leniusculus; Wang R et al.; The prophenoloxidase activating enzyme (ppA), a serine proteinase catalyzing the conversion of prophenoloxidase to an active phenoloxidase, has a molecular mass of about 36 kDa in its active form . This protein was cloned from a blood cell cDNA library and its corresponding cDNA of 1736 base pairs encodes a zymogenic protein (proppA) of 468 amino acids . An antibody raised against a synthetic peptide derived from a region of the cDNA sequence could efficiently inhibit the beta-1,3-glucan triggered activation of prophenoloxidase in vitro . The C-terminal half of the proppA is composed of a typical serine proteinase domain, with a sequence similar to other invertebrate and vertebrate serine proteinases . The N-terminal half contains a cationic glycine-rich domain, a cationic proline-rich domain and a clip-domain, in which the disulfide-bonding pattern is likely to be identical to those of the horseshoe crab big defensin and mammalian beta-defensins . Antibodies made against both the C- and the N-terminal halves recognize two proppAs under reducing conditions . However, under nonreducing conditions only the anti-C antibody recognized the two proppAs, which suggests that a conformational change takes place upon reduction that allows the anti-N to react with the N-terminal half of proppA . The recombinant clip-domain in crayfish proppA was overexpressed in Escherichia coli and the resulting peptide exhibited antibacterial activity against Gram-positive bacterial strains such as Micrococcus luteus Ml11 and Bacillus megaterium Bm11 with 50% growth inhibitory concentrations of 1.43 microM and 17.9 microM, respectively . These results suggest that the clip-domains in proppAs may function as antibacterial peptides. Infect Immun, 2001 Mar, 69(3), 1847 - 55 Effects of tumor necrosis factor alpha on host immune response in chronic persistent tuberculosis: possible role for limiting pathology; Mohan VP et al.; Reactivation of latent tuberculosis contributes significantly to the incidence of disease caused by Mycobacterium tuberculosis . The mechanisms involved in the containment of latent tuberculosis are poorly understood . Using the low-dose model of persistent murine tuberculosis in conjunction with MP6-XT22, a monoclonal antibody that functionally neutralizes tumor necrosis factor alpha (TNF-alpha), we examined the effects of TNF-alpha on the immunological response of the host in both persistent and reactivated tuberculous infections . The results confirm an essential role for TNF-alpha in the containment of persistent tuberculosis . TNF-alpha neutralization resulted in fatal reactivation of persistent tuberculosis characterized by a moderately increased tissue bacillary burden and severe pulmonic histopathological deterioration that was associated with changes indicative of squamous metaplasia and fluid accumulation in the alveolar space . Analysis of pulmonic gene and protein expression of mice in the low-dose model revealed that nitric oxide synthase was attenuated during MP6-XT22-induced reactivation, but was not totally suppressed . Interleukin-12p40 and gamma interferon gene expression in TNF-alpha-neutralized mice was similar to that in control mice . In contrast, interleukin-10 expression was augmented in the TNF-alpha-neutralized mice . In summary, results of this study suggest that TNF-alpha plays an essential role in preventing reactivation of persistent tuberculosis, modulates the pulmonic expression of specific immunologic factors, and limits the pathological response of the host. Infect Immun, 2001 Mar, 69(3), 1704 - 7 Immune response to infection with Mycobacterium ulcerans; Gooding TM et al.; Mycobacterium ulcerans is a slow-growing, acid-fast bacillus that causes chronic necrotizing skin ulcers known as Buruli ulcers . Previously reported information on immunity to this mycobacterium is limited . We examined immune responses to M . ulcerans and M . bovis BCG in patients with M . ulcerans disease and in 20 healthy control subjects (10 tuberculin test positive and 10 tuberculin test negative) . Cell-mediated immunity was assessed by stimulating peripheral blood mononuclear cells (PBMC) with whole mycobacteria and then measuring PBMC proliferation and the production of gamma interferon (IFN-gamma) . Humoral immunity was assessed by immunoblotting . PBMC from all subjects showed significantly greater proliferation and IFN-gamma production in response to stimulation with living mycobacteria compared with killed cells . However, PBMC from subjects with past or current M . ulcerans disease showed significantly reduced proliferation and production of IFN-gamma in response to stimulation with live M . ulcerans or M . bovis than PBMC from healthy, tuberculin test-positive subjects (P < 0.001) and showed results in these assays comparable to those of tuberculin test-negative subjects (P > 0.2) . Serum from 9 of 11 patients with M . ulcerans disease, but no control subject, contained antibodies to M . ulcerans . The results indicate that patients with M . ulcerans infection mount an immune response to M . ulcerans as evidenced by antibody production, but they demonstrate profound systemic T-cell anergy to mycobacterial antigens . These findings may explain some of the distinct clinical and pathological features of M . ulcerans-induced disease. Biochem Biophys Res Commun, 2001 Feb 16, 281(1), 151 - 8 Molecular cloning of an alpha-glucosidase-like gene from Penicillium minioluteum and structure prediction of its gene product; Garcia B et al.; The dexC cDNA, which is expressed in dextran-containing medium by the filamentous fungus Penicillium minioluteum, was cloned and sequence characterized . The cDNA sequence comprises 1859 bp plus a poly (A) tail, coding for a predicted protein of 597 amino acids . The genomic counterpart was isolated by PCR, finding three introns in its sequence . The dexC gene was located by Southern blot in the same 9-kb fragment that the previously isolated dextranase-encoding gene (dexA) . Sequence analysis revealed that the deduced DexC protein belongs to glycosyl hydrolase family 13, showing a high sequence identity (58%) with Aspergillus parasiticus alpha-1,6-glucosidase . In addition, the high sequence identity (51%) between DexC protein and oligo-1,6-glucosidase of Bacillus cereus, with three-dimensional (3D) structure determined, leads us to proposed a 3D model for the structural core of DexC protein. Curr Microbiol, 2001 Apr, 42(4), 237 - 40 Cloning and characterization of a bacterial cell-bound type B carboxylesterase from Bacillus sp . BP-7; Prim N et al.; A clone producing halos on tributyrin plates was isolated from a genomic library of Bacillus sp . BP-7 . The insert contained an open reading frame that coded for a protein of 487 amino acids with homology to carboxylesterases . The cloned enzyme showed clear preference for esters of short-chain fatty acids, being classified as an esterase . Maximum activity was found at 45 degrees C and pH 7.5 . The enzyme displayed stability in the pH range from 6 to 9.5, and at temperatures from 4 degrees to 45 degrees C . Zymogram analysis of the protein revealed a molecular mass of 53 kDa and a pI of 5.1 . The enzyme showed homology to members of the bacterial subclass of type B carboxylesterases, a set of proteins potentially useful for biotechnological applications. J Urol, 2001 Mar, 165(3), 834 - 6 Primary treatment of condylomata acuminata with viable bacillus Calmette-Guerin; Bohle A et al.; PURPOSE: Condylomata acuminata or genital warts are caused by human papillomavirus . Prevalence data show that infection rates are increasing . To our knowledge we report the first successful primary treatment of genital warts with topical bacillus Calmette-Guerin (BCG) and provide long-term followup in a group of adjuvant treated patients with recurrent condylomata acuminata . MATERIALS AND METHODS: In 10 consecutive men viable BCG was directly applied to the condylomata acuminata lesions once weekly for 6 weeks . In nonresponding patients another course of 9 applications was administered for 3 weeks . RESULTS: A complete response was achieved in 6 of the 10 men after 1 or 2 treatment cycles . All responding patients are disease-free at a median followup of 9.2 months (range 4 to 12) . One patient achieved partial regression of the lesions and in 3 the condylomata did not disappear . Side effects were rare and mild . Long-term followup in 6 adjuvant treated patients with rapidly recurrent condylomata acuminata showed no further recurrence after topical BCG in 5 at a median of 30.8 months (range 29 to 50) . CONCLUSIONS: Topical application of viable BCG has therapeutic activity as adjuvant and primary treatment for penile condylomata acuminata with negligible side effects . Long-term followup implies the prevention of recurrent disease. J Urol, 2001 Mar, 165(3), 745 - 56 The limits of bacillus Calmette-Guerin for carcinoma in situ of the bladder; Kim JC et al.; PURPOSE: Historically carcinoma in situ of the bladder has been treated with radical cystectomy based on the aggressive and potentially invasive nature of this disease . The introduction in the late 1970s of intravesical bacillus Calmette-Guerin (BCG) has made this therapy the gold standard in the management of carcinoma in situ . Cases that are refractory or resistant to BCG therapy are a management dilemma with various available treatment options . MATERIALS AND METHODS: A comprehensive literature review of the current management of carcinoma in situ of the bladder was performed using MEDLINE, a review of current urology journals and abstracts from recent urology meetings . Data focused on BCG resistant carcinoma in situ of the bladder and current approaches in use for refractory disease . RESULTS: Complete and durable response rates have been reported in more than 70% of patients with carcinoma in situ who are treated with intravesical BCG . To our knowledge the optimal therapeutic regimen has not been established, although extended periods of treatment beyond the originally described 6-week course have not been shown to improve complete response rates . Prolonged administration of BCG is associated with adverse side effects . Various prognostic indicators of recurrence and progression exist that may identify a subset of cases unlikely to respond favorably to a conservative approach, including carcinoma in situ with associated stage T1 bladder lesions, diffuse and multifocal carcinoma in situ, multiple recurrences with intravesical therapy and extravesical involvement . Current molecular markers may also predict the response of carcinoma in situ to therapy . Treatment options available for BCG refractory carcinoma in situ of the bladder include intravesical chemotherapy, combined immuno-chemotherapy and radical cystectomy . Intravesical valrubicin and oral bropirimine have been shown to induce a complete response rate of 21% to 50%, although data on long-term followup are forthcoming . Radical cystectomy remains effective therapy for aggressive carcinoma in situ of the bladder . CONCLUSIONS: The current management of carcinoma in situ of the bladder is ill defined due to the variable natural history and unpredictable response of this disease to therapy . Controversy exists as to the optimal treatment of carcinoma in situ of the bladder since different forms of carcinoma in situ may exist that complicate therapeutic decisions for appropriate therapy . Some tumor characteristics are associated with more aggressive behavior and may be predictive of treatment outcome. Curr Opin Pediatr, 2001 Feb, 13(1), 56 - 9 Treatment of cat-scratch disease; Conrad DA; Cat-scratch disease is an infection caused by Bartonella henselae, a fastidious gram-negative bacillus acquired from exposure to an infected kitten or cat . The most common manifestation of human disease is lymphadenitis . Atypical forms of infection include Parinaud oculoglandular syndrome, stellate neuroretinitis, persistent fever without localizing signs, hepatosplenic infection, encephalopathy, osteomyelitis, and endocarditis . Immunocompromised individuals with B . hensalae infection may develop bacillary angiomatosis, bacillary peliosis, and relapsing bacteremia with fever syndrome . The bacillus is susceptible to several antibacterial agents in vitro, including penicillins, cephalosporins, aminoglycosides, tetracyclines, macrolides, quinolones, trimethoprim and sulfamethoxazole, and rifampin . Greatest clinical efficacy has been observed following treatment with rifampin, ciprofloxacin, gentamicin, trimethoprim and sulfamethoxazole, clarithromycin, and azithromycin . In one placebo-controlled study, azithromycin therapy was associated with more rapid diminution in size of infected lymph nodes . The majority of cases of cat-scratch disease occurring in normal hosts do not require anti-infective therapy for resolution of infection. J Am Acad Dermatol, 2001 Feb, 44(2), 261 - 4 Utility of anti-bacillus Calmette-Guérin antibodies as a screen for organisms in sporotrichoid infections; Byrd J et al.; BACKGROUND: Sporotrichoid infections often present a diagnostic challenge for the dermatopathologist . Therefore an affordable, reliable method of easier recognition of organisms is desired . Recently, immunohistochemical staining with antibodies against Mycobacterium bovis (bacillus Calmette-Guerin, BCG) was found to be useful in identifying various fungi and mycobacteria with minimal background staining . OBJECTIVE: The purpose of this study was to demonstrate the usefulness of anti-BCG antibodies as a screen for organisms in sporotrichoid infections . METHODS: Thirteen specimens of suspected sporotrichosis were selected for staining with anti-BCG antibody . Sporotrichoid infection was confirmed by histochemical staining, biopsy, and follow-up results . RESULTS: Twelve of the 13 specimens stained positively using anti-BCG antibody . Of the 5 cultures done, 2 were positive for M . marinum, and 1 grew Sporothrix schenckii . CONCLUSION: Immunohistochemical staining with anti-BCG antibody offers another screening method to identify organisms in sporotrichoid infections because of its ease, cost-effectiveness, and simplicity. J Biochem (Tokyo), 2001 Feb, 129(2), 303 - 12 Crystal structure of glucose dehydrogenase from Bacillus megaterium IWG3 at 1.7 A resolution; Yamamoto K et al.; The crystal structure of glucose dehydrogenase (GlcDH) from Bacillus megaterium IWG3 has been determined to an R-factor of 17.9% at 1.7 A resolution . The enzyme consists of four identical subunits, which are similar to those of other short-chain reductases/dehydrogenases (SDRs) in their overall folding and subunit architecture, although cofactor binding sites and subunit interactions differ . Whereas a pair of basic residues is well conserved among NADP(+)-preferring SDRs, only Arg39 was found around the adenine ribose moiety of GlcDH . This suggests that one basic amino acid is enough to determine the coenzyme specificity . The four subunits are interrelated by three mutually perpendicular diad axes (P, Q, and R) . While subunit interactions through the P-axis for GlcDH are not so different from those of the other SDRs, those through the Q-axis differ significantly . GlcDH was found to have weaker hydrophobic interactions in the Q-interface . Moreover, GlcDH lacks the salt bridge that stabilizes the subunit interaction in the Q-interface in the other SDRs . Hydrogen bonds between Q-axis related subunits are also less common than in the other SDRs . The GlcDH tetramer dissociates into inactive monomers at pH 9.0, which can be attributed mainly to the weakness of the Q-axis interface. Acta Crystallogr D Biol Crystallogr, 2001 Feb, 57(Pt 2), 292 - 5 Crystallization and preliminary X-ray analysis of the sporulation factor SpoIIAA in its native and phosphorylated forms; Seavers PR et al.; Sporulation in Bacillus begins with an asymmetric cell division producing two progeny with identical chromosomes but different developmental fates . As such, it is a simple example of cellular differentiation . The establishment of cell type is controlled by a series of alternate RNA polymerase sigma subunits . The first compartment-specific sigma factor is sigma(F), whose activity is controlled by SpoIIAB, an anti-sigma factor, and SpoIIAA, an anti-sigma factor antagonist which is phosphorylated by the kinase activity of SpoIIAB . Here, the preliminary crystallographic analysis of SpoIIAA and phosphorylated SpoIIAA from B . sphaericus in forms suitable for high-resolution structure determination are reported. FEBS Lett, 2001 Feb 9, 490(1-2), 70 - 4 The role of a proline-induced broken-helix motif in alpha-helix 2 of Bacillus thuringiensis delta-endotoxins; Arnold S et al.; Bacillus thuringiensis delta-endotoxins (Cry proteins), are widely used for insect control and plant protection . They are water-soluble proteins that insert into membranes forming ion channels . In most Cry toxins alpha-helix 2 is broken by a highly conserved proline residue (Pro70 in Cry1Ab), generating a broken-helix motif . The flexibility of the motif was altered through site-directed mutagenesis . It was found that increasing the flexibility of the motif decreased the stability, the ion transport ability and the toxicity of the protein . By removing the broken-helix motif, the biological properties were restored to a wild type level. Environ Health Perspect, 2001 Jan, 109(1), 47 - 54 Spatial and temporal distribution of airborne Bacillus thuringiensis var . kurstaki during an aerial spray program for gypsy moth eradication; Teschke K et al.; We measured airborne exposures to the biological insecticide Bacillus thuringiensis var . kurstaki (Btk) during an aerial spray program to eradicate gypsy moths on the west coast of Canada . We aimed to determine whether staying indoors during spraying reduced exposures, to determine the rate of temporal decay of airborne concentrations, and to determine whether drift occurred outside the spray zone . During spraying, the average culturable airborne Btk concentration measured outdoors within the spray zone was 739 colony-forming units (CFU)/m3 of air . Outdoor air concentrations decreased over time, quickly in an initial phase with a half time of 3.3 hr, and then more slowly over the following 9 days, with an overall half-time of about 2.4 days . Inside residences during spraying, average concentrations were initially 2-5 times lower than outdoors, but at 5-6 hr after spraying began, indoor concentrations exceeded those outdoors, with an average of 244 CFU/m3 vs . 77 CFU/m3 outdoors, suggesting that the initial benefits of remaining indoors during spraying may not persist as outside air moves indoors with normal daily activities . There was drift of culturable Btk throughout a 125- to 1,000-meter band outside the spray zone where measurements were made, a consequence of the fine aerosol sizes that remained airborne (count median diameters of 4.3 to 7.2 microm) . Btk concentrations outside the spray zone were related to wind speed and direction, but not to distance from the spray zone. J Pharm Sci, 2001 Mar, 90(3), 275 - 87 Thermal sterilization of heat-sensitive products using high-temperature short-time sterilization; Mann A et al.; High-temperature short-time (HTST) sterilization with a continuous-flow sterilizer, developed for this study, was evaluated . The evaluation was performed with respect to (a) the chemical degradation of two heat-sensitive drugs in HTST range (140-160 degrees C) and (b) the microbiological effect of HTST sterilization . Degradation kinetics of two heat-sensitive drugs showed that a high peak temperature sterilization process resulted in less chemical degradation for the same microbiological effect than a low peak temperature process . Both drugs investigated could be sterilized with acceptable degradation at HTST conditions . For the evaluation of the microbiological effect, Bacillus stearothermophilus ATCC 7953 spores were used as indicator bacteria . Indicator spore kinetics (D(T), z value, k, and E(a)), were determined in the HTST range . A comparison between the Bigelow model (z value concept) and the Arrhenius model, used to describe the temperature coefficient of the microbial inactivation, demonstrated that the Bigelow model is more accurate in prediction of D(T) values in the HTST range . The temperature coefficient decreased with increasing temperature . The influence of Ca(2+) ions and pH value on the heat resistance of the indicator spores, which is known under typical sterilization conditions, did not change under HTST conditions. Biopolymers, 2001 Mar, 58(3), 260 - 7 Conformational characterization of designed minibarnase; Takahashi K et al.; We have designed a minibarnase by removing one module from barnase, a bacterial RNase from Bacillus amyloliquefaciens . Barnase, consisting of 110 amino acid residues, is decomposed into six modules, M1-M6 . Module is defined as a peptide segment consisting of contiguous amino acid residues that makes a small compact conformation within a globular domain . To understand the role of module in protein architecture, we analyzed NMR and CD spectra of a minibarnase, which lacked 26 amino acid residues corresponding to module M2 . We demonstrated the formation of hydrophobic cores in the minibarnase similar to those of barnase . Although its conformational stability against acids and heat was reduced in comparison with barnase, the minibarnase retained cooperative folding character (two-state folding) . Therefore, the folding of the minibarnase consisting of modules M1 and M3-M6 is independent to some extent of module M2 . This finding may be useful for future module-based protein design . Mol Microbiol, 2001 Feb, 39(3), 813 - 21 An increase in expression of a Mycobacterium tuberculosis mycolyl transferase gene (fbpB) occurs early after infection of human monocytes; Wilkinson RJ et al.; Changes in the mRNA levels of two Mycobacterium tuberculosis genes (fbpB known as antigen 85B, and hspX known as Acr) were studied in infected human monocytes . Antigen 85B is an enzyme involved in cell wall biosynthesis and is also a major target of the immune response . Acr is a stress protein believed to be involved in the bacillary response to adverse conditions and in non-replicating persistence . During the first 24 h of intracellular infection, the intramonocyte 85B mRNA level increased 54-fold (P = 0.00001) and 14.6 times in comparison with the 16S ribosomal rRNA . In contrast, the Acr mRNA fell 14.3 times . Although monocyte cytokine production was very variable, the 24 h secretion of tumour necrosis factor (TNF)-alpha correlated with the 85B-16S RNA ratio at 24 h (r = 0.77, Pcorr < 0.01) . Furthermore, the addition of exogenous TNF-alpha to cultures was associated with a twofold increase in the 85B-16S ratio and, conversely, neutralization of endogenous TNF-alpha reduced the ratio . As antigen 85B also induces TNF-alpha, the positive feedback implied by our findings suggests a previously unsuspected role for this protein in the immunopathogenesis of tuberculosis. Environ, Toxicol . Pharmacol. . 2001 Jan 1, 9(3), 79 - 85 Domoic acid: a fascinating marine toxin; Mos L; There are indications that toxic algal blooms are increasing because of pollution of coastal waters and worldwide shipping . This mini-review deals with the marine biotoxin domoic acid, also known as amnesic shellfish poison, and its main producing pennate diatom genus Pseudo-nitzschia (Bacillariophyceae) . Besides contamination of seafood, these organisms have also been involved in human and marine wildlife mortality . The article aims to give an overview of all biological and environmental factors that should be considered when trying to evaluate a possible increase in toxic blooms of Pseudo-nitzschia spp . Pseudo-nitzschia blooms characteristically occur in a low light regime, at a time when the temperature is falling and at a wide range of salinities . Laboratory studies have shown that the production of domoic acid, a water-soluble amino acid, is related to silicon, phosphorus, nitrogen and trace metal (mainly iron) availability . Domoic acid has no known function in defence or primary metabolism; a role in excretion of excess photosynthetic energy or as a binding ligand for trace metals is suggested . The variability in domoic acid production by different Pseudo-nitzschia spp., or the presence of toxic and non-toxic strains of the same species, cannot be explained . The conclusion is drawn that an increase in toxic blooms of Pseudo-nitzschia spp . might be possible, especially because of the expected increase in nutrient availability from pollution and desert dust . Global warming may have an influence as well by lengthening the growth period for Pseudo-nitzschia, enlarging their global distribution and increasing the dust load through desertification. FEMS Microbiol Lett, 2001 Feb 5, 195(1), 67 - 72 Identification of thermostabilizing residues in a Bacillus alkaline cellulase by construction of chimeras from mesophilic and thermostable enzymes and site-directed mutagenesis; Hakamada Y et al.; An alkaliphilic Bacillus sp . strain, KSM-64, produces a mesophilic alkaline endo-1,4-beta-glucanase that is suitable for use in detergents . The deduced amino acid sequence of the enzyme showed very high homology to that of a thermostable alkaline enzyme from alkaliphilic Bacillus sp . strain KSM-S237 . Analysis of chimeric enzymes produced from the genes encoding the mesophilic and thermostable enzymes suggested that the lysine residues at positions 137, 179, and 194 are responsible for their thermal stabilization . Replacing the corresponding Glu137, Asn179, and/or Asp194 with lysine by site-directed mutagenesis made the mesophilic enzyme more thermostable . Analyses of the hydrophilicity of deduced amino acid sequences and isoelectric focusing of the modified enzymes suggested that these three specific lysine residues and their replacements are all located on the surface of the enzyme molecule . This fact further suggested that specific ionic interaction is involved in the thermal stabilization of the enzyme. FEMS Microbiol Lett, 2001 Feb 5, 195(1), 1 - 8 Why Bacillus thuringiensis insecticidal toxins are so effective: unique features of their mode of action; Aronson AI et al.; The spore-forming bacterium Bacillus thuringiensis produces intracellular inclusions comprised of protoxins active on several orders of insects . These highly effective and specific toxins have great potential in agriculture and for the control of disease-related insect vectors . Inclusions ingested by larvae are solubilized and converted to active toxins in the midgut . There are two major classes, the cytolytic toxins and the delta-endotoxins . The former are produced by B . thuringiensis subspecies active on Diptera . The latter, which will be the focus of this review, are more prevalent and active on at least three orders of insects . They have a three-domain structure with extensive functional interactions among the domains . The initial reversible binding to receptors on larval midgut cells is largely dependent upon domains II and III . Subsequent steps involve toxin insertion into the membrane and aggregation, leading to the formation of gated, cation-selective channels . The channels are comprised of certain amphipathic helices in domain I, but the three processes of insertion, aggregation and the formation of functional channels are probably dependent upon all three domains . Lethality is believed to be due to destruction of the transmembrane potential, with the subsequent osmotic lysis of cells lining the midgut . In this review, the mode of action of these delta-endotoxins will be discussed with emphasis on unique features. Enzyme Microb Technol, 2001 Feb 1, 28(2-3), 161 - 167 trans-Sialidase catalyzed sialylation of beta-galactosyldisaccharide with an introduction of beta-galactosidase; Lee S et al.; Introduction of beta-galactosidase into a trans-sialidase reaction, i.e . sialic acid transfer reaction from a donor substrate (alpha2,3-sialyllactose) to an acceptor substrate (beta-galactosyldisaccharide), could improve the yield of desired sialylated trisaccharide by hydrolyzing lactose, a byproduct from the donor . When trans-sialidase reaction was performed with stoichiometric amounts (2 mM) of alpha2,3-sialyllactose and Galbeta(1,3)GlcNAc, the yield of NeuAcalpha(2,3)Galbeta(1,3)GlcNAc increased from 45% to 75% by the coupling of Escherichia coli beta-galactosidase . Furthermore, by changing the substrate ratio in the coupled reaction, i.e . two-fold excess of alpha2,3-sialyllactose to Galbeta(1,3)GlcNAc, above 95% of yield was achieved based on the amount of Galbeta(1,3)GlcNAc . However, two-fold excess of Galbeta(1,3)GlcNAc to alpha2,3-sialyllactose in this reaction was more desirable for the purification of NeuAcalpha(2,3)Galbeta(1,3)GlcNAc, since complete consumption of alpha2,3-sialyllactose was achieved . Efficiency of the coupled reaction was affected by the specificity of beta-galactosidase for acceptor substrate . When Galbeta(1,6)GlcNAc was used as the acceptor, E . coli beta-galactosidase hydrolyzed Galbeta(1,6)GlcNAc as well as lactose in the coupled reaction, resulting in a significant decrease in the yield of desired sialylated trisaccharide . The conversion yield of the sialylation of Galbeta(1,6)GlcNAc could be improved by employing Bacillus circulans beta-galactosidase. Neth J Med, 2001 Feb, 58(2), 71 - 5 BCG immunotherapy: be cautious of granulomas . Disseminated BCG infection and mycotic aneurysm as late complications of intravesical BCG instillations; Kamphuis JT et al.; We describe a 65-year-old man with a granulomatous hepatitis and a progressive mycotic aneurysm of the abdominal aorta . One year before he received intravesical bacillus Calmette--Guerin (BCG) for carcinoma of the bladder without any complaints . Only post-mortem investigations could confirm that he suffered from a systemic BCG infection . Literature is reviewed for this rare complication. Toxicology, 2001 Jan 2, 156(2-3), 101 - 7 Bacillus intermedius ribonuclease as inhibitor of cell proliferation and membrane current; Ilinskaya O et al.; The antiproliferative action of the guanine-specific ribonuclease secreted by Bacillus intermedius (binase) was studied in different chicken and mouse cell lines . The proliferation rate of chicken embryo fibroblasts, either normal or Rous sarcoma virus-transformed, was significantly reduced by binase treatment . Among mouse fibroblasts, v-ras-transformed NIH3T3 cells were sensitive to binase, whereas the growth of non-transformed, v-src-transformed or v-fms-transformed NIH3T3 cells was not affected . A 48 h treatment with binase inhibited the Ca2+-dependent K+ current of v-ras-transformed NIH3T3 cells but had no effect on this membrane current in non-transformed and in v-src- or v-fms-transformed NIH3T3 cells . Our results suggest that mammalian cells expressing the ras-oncogene are a potential target for the antiproliferative action of binase. J Biotechnol, 2000 Dec 28, 84(3), 249 - 57 P450 in biotechnology: zinc driven omega-hydroxylation of p-nitrophenoxydodecanoic acid using P450 BM-3 F87A as a catalyst; Schwaneberg U et al.; Cytochrome P450 enzymes require the delivery of two electrons to the heme protein for their enzymatic function . NADPH or NADH are usually used as reduction equivalents . In the absence of a substrate, NADPH may inactivate P450 enzymes . Furthermore, it is expensive, making it unsuitable for the preparative synthesis of fine chemicals . Approaches for replacing NADPH with an electrochemically generated reduction by using platinum-electrodes and different mediators are known . In the present study, NADPH was substituted by the mediator cobalt(III)sepulchrate and zinc dust that serves as an electron source . The mutated fatty acid hydroxylase P450 BM-3 F87A from Bacillus megaterium was chosen as a catalyst, since it shows a three-fold higher sensitivity and a nearly five-fold higher activity for p-nitrophenoxydodecanoic acid (12-pNCA) than the wild-type enzyme . The formation of p-nitrophenolate can easily be monitored using a photometer at 410 nm . The turnover rate of the zinc/cobalt(III)sepulchrate system reaches 20% of the NADPH activity . Compared to the electrochemical approaches the activity is at least 77% higher (turnover 125 eq min-1) . The presented alternative cofactor system can be used instead of NADPH or expensive electrochemical devices (platinum electrodes) for fine chemical synthesis. Vaccine, 2001 Jan 8, 19(11-12), 1460 - 6 Mucosal immunization against hepatitis B virus by intranasal co-administration of recombinant hepatitis B surface antigen and recombinant cholera toxin B subunit as an adjuvant; Isaka M et al.; Recombinant cholera toxin B subunit (rCTB) produced by Bacillus brevis carrying pNU212-CTB has been previously found to be a potent mucosal adjuvant to aluminium-non-adsorbed tetanus toxoid (nTT) and diphtheria toxoid (nDT) co-administered intranasally, and the possibility of needle-free inoculation of these vaccines with rCTB has been suggested . In this paper we examined the potentiality of rCTB as a mucosal adjuvant to aluminium-non-adsorbed yeast-derived recombinant hepatitis B surface antigen (rHBs) being a particulate antigen when administered intranasally with rCTB . In-house ELISA showed that a mixture of rHBs (1 or 5 microg) and rCTB (10 microg) elevated not only systemic responses but also mucosal immune responses at the nasal cavity, the lung, the saliva, the small intestine and the vagina against rHBs, and these could be further increased with higher doses of antigen . With antibody isotypes of IgG, there were equally high levels of serum HBs-specific IgG1, IgG2a and IgG2b antibodies and induction of mixed Th1- and Th2-type responses was considered to occur in combination of rHBs and rCTB . Serum anti-HBs titres in almost all mice obtained from sandwich EIA using a commercial kit were higher than 1000 milli-international units ml(-1) (mIU ml(-1)) . These results show that rCTB is also very effective as a mucosal adjuvant for a particulate antigen like rHBs, as well as soluble antigens like nTT and nDT reported previously, suggesting the possibility of intranasal immunization with rHBs plus rCTB in humans. Vaccine, 2001 Jan 8, 19(11-12), 1391 - 6 DNA encoding a single mycobacterial antigen protects against leprosy infection; Martin E et al.; The continuing incidence of leprosy infection around the world and the inability of Mycobacterium bovis bacille Calmette-Guerin (BCG) to protect certain populations clearly indicates that an improved vaccine against leprosy is needed . The immuno dominant 35 kDa protein, shared by Mycobacterium leprae and Mycobacterium avium, but not Mycobacterium tuberculosis or BCG, is recognised by >90% of leprosy patients, making it an ideal candidate antigen for a subunit vaccine . Immunization of outbred Swiss Albino mice with a DNA-35 vaccine stimulated specific T cell activation and IFN-gamma production . DNA-35 immunization induced significant levels of protection against M . leprae footpad infection, comparable to that produced by BCG . Therefore, DNA immunization with the 35 kDa antigen is effective against M . leprae infection and genetic immunization with a combination of antigens holds the potential for an improved vaccine against leprosy. Mol Immunol, 2000 Jun, 37(9), 527 - 36 Co-expression of interleukin-2 and green fluorescent protein reporter in mycobacteria: in vivo application for monitoring antimycobacterial immunity; Luo Y et al.; Recombinant Mycobacterium bovis bacillus Calmette-Guerin expressing green fluorescent protein (rBCG-GFP), driven by the mycobacterial heat shock protein 70 (HSP 70) promoter from an autonomously replicating plasmid, was genetically engineered to co-express mouse interleukin-2 (IL-2) by introduction of an independent HSP 60 promoter . To monitor host antimycobacterial immunity, C57BL/6 mice were intravenously infected with IL-2 expressing and non-expressing GFP rBCGs . Both rBCGs were clearly imaged and easily quantified with ultraviolet microscopy of tissue sections and whole organ suspensions . Enhanced mycobacterial clearance from the spleens of mice infected with the rBCG-IL-2/GFP strain was apparent by both diminished bacterial counts and spleen weights during the first 6 weeks post-infection relative to rBCG-GFP . T helper type 1 (TH1) cytokine production and proliferative response to BCG restimulation was also elevated from in vitro splenocyte cultures taken from the rBCG-IL-2/GFP-infected group . Taken together, these results suggest that IL-2 expression from rBCG augmented host protective immunity to mycobacterial infection via an enhanced TH1 immune response . Mycobacterial expression vectors that allow simultaneous but independent production of reporter proteins and bioactive substances provide an ideal means for monitoring the in vivo fate of recombinant mycobacteria. Mol Cell Endocrinol, 2000 Dec 22, 170(1-2), 163 - 74 Single-column purification and bio-characterization of recombinant human parathyroid hormone-related protein (1-139); Wu C et al.; Recombinant human parathyroid hormone-related protein (hPTHrP) (1-139) was expressed using the IMPACT T7 (intein-mediated purification with an affinity chitin-binding tag) system, allowing purification of free recombinant peptide in a single chromatographic step . This system utilizes an intein, which is a protein splicing element from the Saccharomyces cerevisiae VMA1 gene . The intein has been modified so that it undergoes a self-cleavage reaction at its N-terminus at low temperatures in the presence of 1,4-dithiothreitol (DTT) . The cDNA encoding hPTHrP (1-139) was cloned into the pTYB1 vector to create an in-frame fusion at the N-terminus of the intein gene . The cDNA for the chitin-binding domain from Bacillus circulans is present at the C-terminus of intein for affinity purification of the three-part fusion protein on a chitin column . The recombinant plasmid was transfected into E . coli ER2566 cells and synthesis of the PTHrP fusion protein was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG) . This system produced pure hPTHrP (1-139) and an N-terminally truncated analogue, hPTHrP (27-139), as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blot analysis, N-terminal sequence analysis and mass spectroscopy . hPTHrP (1-139) stimulated cAMP accumulation in ROS 17/2.8 osteoblastic bone cells, whereas hPTHrP (27-139) failed to elicit a response . hPTHrP (1-139) also inhibited the growth of the breast cancer cell line MDA-MB-231; the magnitude of the response was comparable with that of synthetic hPTHrP (1-34) and (1-86) . Neutralization of endogenous PTHrP and added hPTHrP (1-139) and N-terminal species with an anti-PTHrP antiserum completely abolished the growth inhibitory effects . These results indicate that the added peptides modulate cell growth by acting at the cell surface . Availability of recombinant hPTHrP (1-139) will allow further study of its biological function, as well as its structure. Biochem Biophys Res Commun, 2001 Feb 2, 280(4), 1177 - 82 Fragmentary form of thermostable leucine dehydrogenase of Bacillus stearothermophilus: its construction and reconstitution of active fragmentary enzyme; Oikawa T et al.; X-ray crystallographic studies revealed that various amino acid dehydrogenases fold into two domains in each subunit, a substrate-binding domain and an NAD(P)(+)-binding domain (Baker, P . J., Turnbull, A . P., Sedelnikova, S . E., Stillman, T . J., and Rice, D . W . (1995) Structure 3, 693-705) . To elucidate the function and folding process of these two domains, we have genetically constructed a fragmentary form of thermostable leucine dehydrogenase of Bacillus stearothermophilus consisting of an N-terminal polypeptide fragment corresponding to the substrate-binding domain including an N-terminus, and a C-terminal fragment corresponding to the NAD(+)-binding domain . The two peptide fragments were expressed in separate host cells and purified . When both fragments were mixed, the leucine dehydrogenase activity with a specific activity of 1.4% of that of the wild-type enzyme appeared . This suggests that both peptide fragments mutually recognize each other, associate and fold correctly to be catalytically active, although the activity is low . However, the fragmentary form of enzyme produced catalyzed the oxidative deamination of l-leucine, l-isoleucine, and l-valine with broad substrate specificity compared to that of the wild-type enzyme . The fragmentary enzyme retained more than 75% of the initial activity after heating at 50 degrees C for 60 min . The fragmentary enzyme was more stable on heating than separate peptide fragments . These results suggest that the two domains of leucine dehydrogenase probably fold independently, and the two peptide fragments interact and associate with each other to form a functional active site . Nucleic Acids Res, 2001 Feb 15, 29(4), 895 - 903 Characterization of BseMII, a new type IV restriction-modification system, which recognizes the pentanucleotide sequence 5'-CTCAG(N)(10/8)/; Jurenaite-Urbanaviciene S et al.; We report the properties of the new BseMII restriction and modification enzymes from Bacillus stearothermophilus Isl 15-111, which recognize the 5'-CTCAG sequence, and the nucleotide sequence of the genes encoding them . The restriction endonuclease R.BseMII makes a staggered cut at the tenth base pair downstream of the recognition sequence on the upper strand, producing a two base 3'-protruding end . Magnesium ions and S:-adenosyl-L-methionine (AdoMet) are required for cleavage . S:-adenosylhomocysteine and sinefungin can replace AdoMet in the cleavage reaction . The BseMII methyltransferase modifies unique adenine residues in both strands of the target sequence 5'-CTCAG-3'/5'-CTGAG-3' . Monomeric R.BseMII in addition to endonucleolytic activity also possesses methyltransferase activity that modifies the A base only within the 5'-CTCAG strand of the target duplex . The deduced amino acid sequence of the restriction endonuclease contains conserved motifs of DNA N6-adenine methylases involved in S-adenosyl-L-methionine binding and catalysis . According to its structure and enzymatic properties, R.BseMII may be regarded as a representative of the type IV restriction endonucleases. J Bacteriol, 2001 Mar, 183(5), 1672 - 9 S-layer variation in Bacillus stearothermophilus PV72 is based on DNA rearrangements between the chromosome and the naturally occurring megaplasmids; Scholz HC et al.; Bacillus stearothermophilus PV72 expresses different S-layer genes (sbsA and sbsB) under different growth conditions . No stretches of significant sequence identity between sbsA and sbsB were detected . In order to investigate S-layer gene regulation in B . stearothermophilus PV72, we characterized the upstream regulatory region of sbsA and sbsB by sequencing and primer extension analysis . Both genes are transcribed from unique but different promoters, independently of the growth phase . Localization of sbsB in the sbsA-expressing strain PV72/p6 revealed that the coding region of the second S-layer gene sbsB is located not on the chromosome but on a natural megaplasmid of the strain, whereas the upstream regulatory region of sbsB was exclusively detected on the chromosome of PV72/p6 . For sbsB expression, the coding region has to be integrated into the chromosomally located expression site . After the switch to sbsB expression, the sbsA coding region was removed from the chromosome but could still be detected on the plasmid of the sbsB-expressing strain PV72/p2 . The sbsA upstream regulatory region, however, remained on the chromosome . This is the first report of S-layer variation not caused by intrachromosomal DNA rearrangements, but where variant formation depends on recombinational events between the plasmid and the chromosome. Appl Environ Microbiol, 2001 Feb, 67(2), 995 - 1000 Molecular characterization of cycloinulooligosaccharide fructanotransferase from Bacillus macerans; Kim HY et al.; Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cyclooligosaccharides of beta-(2-->1)-linked D-fructofuranose by catalyzing an intramolecular transfructosylation reaction . The CFTase gene was cloned and characterized from Bacillus macerans CFC1 . The CFTase gene encoded a polypeptide of 1,333 amino acids with a calculated Mr of 149,563 . Western blot and zymography analyses revealed that the CFTase with a molecular mass of 150 kDa (CFT150) was processed (between Ser389 and Phe390 residue) to form a 107-kDa protein (CFT107) in the B . macerans CFC1 cells . The processed CFT107 was similar in its mass to the previously purified CFTase from B . macerans CFC1 . The CFT107 enzyme was produced by B . macerans CFC1 but was not detected from the recombinant Escherichia coli cells, indicating that the processing event occurred in a host-specific manner . The two CFTases (CFT150 and CFT107) exhibited the same enzymatic properties, such as influences of pH and temperature on the enzyme activity, the intermolecular transfructosylation ability, and the ability of hydrolysis of cycloinulooligosaccharides produced by the cyclization reaction . However, the thermal stability of CFT107 was slightly higher than that of CFT150 . The most striking difference between the two enzymes was observed in their Km values; the value for CFT150 (1.56 mM) was threefold lower than that for CFT107 (4.76 mM) . Thus, the specificity constant (kcat/Km) of CFT150 was about fourfold higher than that of CFT107 . These results indicated that the N-terminal 358-residue region of CFT150 played a role in increasing the enzyme's binding affinity to the inulin substrate. Appl Environ Microbiol, 2001 Feb, 67(2), 979 - 81 Mannose phosphate isomerase isoenzymes in Plutella xylostella support common genetic bases of resistance to Bacillus thuringiensis toxins in Llpidopteran species; Herrero S et al.; A strong correlation between two mannose phosphate isomerase (MPI) isoenzymes and resistance to Cry1A toxins from Bacillus thuringiensis has been found in a Plutella xylostella population . MPI linkage to Cry1A resistance had previously been reported for a Heliothis virescens population . The fact that the two populations share similar biochemical, genetic, and cross-resistance profiles of resistance suggests the occurrence of homologous resistance loci in both species. Appl Environ Microbiol, 2001 Feb, 67(2), 872 - 9 Binding analyses of Bacillus thuringiensis Cry delta-endotoxins using brush border membrane vesicles of Ostrinia nubilalis; Hua G et al.; Transgenic corn expressing the Bacillus thuringiensis Cry1Ab gene is highly insecticidal to Ostrinia nubilalis (European corn borer) larvae . We ascertained whether Cry1F, Cry9C, or Cry9E recognizes the Cry1Ab binding site on the O . nubilalis brush border by three approaches . An optical biosensor technology based on surface plasmon resonance measured binding of brush border membrane vesicles (BBMV) injected over a surface of immobilized Cry toxin . Preincubation with Cry1Ab reduced BBMV binding to immobilized Cry1Ab, whereas preincubation with Cry1F, Cry9C, or Cry9E did not inhibit BBMV binding . BBMV binding to a Cry1F-coated surface was reduced when vesicles were preincubated in Cry1F or Cry1Ab but not Cry9C or Cry9E . A radioligand approach measured 125I-Cry1Ab toxin binding to BBMV in the presence of homologous (Cry1Ab) and heterologous (Cry1Ac, Cry1F, Cry9C, or Cry9E) toxins . Unlabeled Cry1Ac effectively competed for 125I-Cry1Ab binding in a manner comparable to Cry1Ab itself . Unlabeled Cry9C and Cry9E toxins did not inhibit (125)I-Cry1Ab binding to BBMV . Cry1F inhibited (125)I-Cry1Ab binding at concentrations greater than 500 nM . Cry1F had low-level affinity for the Cry1Ab binding site . Ligand blot analysis identified Cry1Ab, Cry1Ac, and Cry1F binding proteins in BBMV . The major Cry1Ab signals on ligand blots were at 145 kDa and 154 kDa, but a strong signal was present at 220 kDa and a weak signal was present at 167 kDa . Cry1Ac and Cry1F binding proteins were detected at 220 and 154 kDa . Anti-Manduca sexta aminopeptidase serum recognized proteins of 145, 154, and 167 kDa, and anti-cadherin serum recognized the 220 kDa protein . We speculate that isoforms of aminopeptidase and cadherin in the brush border membrane serve as Cry1Ab, Cry1Ac, and Cry1F binding proteins. Appl Environ Microbiol, 2001 Feb, 67(2), 713 - 20 Polysaccharide lyase: molecular cloning, sequencing, and overexpression of the xanthan lyase gene of Bacillus sp . strain GL1; Hashimoto W et al.; When grown on xanthan as a carbon source, the bacterium Bacillus sp . strain GL1 produces extracellular xanthan lyase (75 kDa), catalyzing the first step of xanthan depolymerization (H . Nankai, W . Hashimoto, H . Miki, S . Kawai, and K . Murata, Appl . Environ . Microbiol . 65:2520-2526, 1999) . A gene for the lyase was cloned, and its nucleotide sequence was determined . The gene contained an open reading frame consisting of 2,793 bp coding for a polypeptide with a molecular weight of 99,308 . The polypeptide had a signal peptide (2 kDa) consisting of 25 amino acid residues preceding the N-terminal amino acid sequence of the enzyme and exhibited significant homology with hyaluronidase of Streptomyces griseus (identity score, 37.7%) . Escherichia coli transformed with the gene without the signal peptide sequence showed a xanthan lyase activity and produced intracellularly a large amount of the enzyme (400 mg/liter of culture) with a molecular mass of 97 kDa . During storage at 4 degrees C, the purified enzyme (97 kDa) from E . coli was converted to a low-molecular-mass (75-kDa) enzyme with properties closely similar to those of the enzyme (75 kDa) from Bacillus sp . strain GL1, specifically in optimum pH and temperature for activity, substrate specificity, and mode of action . Logarithmically growing cells of Bacillus sp . strain GL1 on the medium with xanthan were also found to secrete not only xanthan lyase (75 kDa) but also a 97-kDa protein with the same N-terminal amino acid sequence as that of xanthan lyase (75 kDa) . These results suggest that, in Bacillus sp . strain GL1, xanthan lyase is first synthesized as a preproform (99 kDa), secreted as a precursor (97 kDa) by a signal peptide-dependent mechanism, and then processed into a mature form (75 kDa) through excision of a C-terminal protein fragment with a molecular mass of 22 kDa. Curr Microbiol, 2000 Jul, 41(1), 65 - 9 Cloning of a new Bacillus thuringiensis cry1I-type crystal protein gene; Choi SK et al.; A new cry1I-type gene, cry1Id1, was cloned from a B . thuringiensis isolate, and its nucleotide sequence was determined . The deduced amino acid sequence of Cry1Id1 is 89.7%, 87.2%, and 83.4% identical to the Cry1Ia, Cry1Ib, and Cry1Ic proteins, respectively . The upstream sequence of the cry1Id1 structural gene was not functional as promoter in B . subtilis . The Cry1Id1 protein, purified from recombinant E . coli cells, had a toxicity comparable to that of Cry1Ia against Plutella xylostella, but it was significantly less active than Cry1Ia against Bombyx mori . Cry1Id1 was not active against the coleopteran insect, Agelastica coerulea. Folia Biol (Krakow), 1999, 47(3-4), 143 - 8 Biocidal activity of Bacillus species for Anopheles larvae; Shakoori AR et al.; It is generally accepted that Bacillus thuringiensis (B.t.) is specifically toxic to some insects but does not pose any threat to the environment, operators, or consumers . There are several other Bacillus species which can be used as effective bioinsecticides . In this study four different species of Bacillus, i.e., B . coagulans, B . megaterium, B . brevis, and B . sphaericus were isolated from soil samples collected from Kala Shah Kakoo and Kasur areas, in the suburbs of Lahore . Isolated Bacillus species were administered to mosquito larvae to evaluate their biocidal activity . B . coagulans I from Kala Shah Kakoo showed 93% mortality, while B . coagulans III from Kasur showed 70% mortality . Bacterial isolates most toxic to Anopheles larvae showed optimum growth at 37 degrees C and pH 7 . These isolates have a great potency to controlling anopheline population. Urology . 1999 Jun;53(6):1228. Granulomatous nephritis as a complication of intrarenal bacille Calmette-Guérin therapy; Soda T et al.; A case of granulomatous nephritis after intrarenal bacille Calmette-Guerin (BCG) therapy is reported . High fever greater than 38.5 degrees C lasted for 1 month, without response to conservative therapy . Standard nephroureterectomy was subsequently carried out . Histopathologic findings from the surgical specimen were compatible with BCG-induced granulomatous nephritis . The use of a syringe pump for retrograde instillation of BCG was thought to be the major cause of this severe complication. Arch Virol, 2000, 145(2), 275 - 89 Inter- and intra-site genetic diversity of natural field populations of rice tungro bacilliform virus in the Philippines; Arboleda M et al.; The genetic structure of rice tungro bacilliform virus (RTBV) populations within and between growing sites was analyzed in a collection of natural field isolates from different rice varieties grown in eight tungro-endemic sites of the Philippines . Total DNA extracts from 345 isolates were digested with EcoRV restriction enzyme and hybridized with a full-length probe of RTBV, a procedure shown in preliminary experiments capable of revealing high levels of polymorphism in RTBV field isolates . In the total population, 17 distinct EcoRV-based genome profiles (genotypes) were identified and used as indicators for virus diversity . Distinct sets of genotypes occurred in Isabela and North Cotabato provinces suggesting a geographic isolation of virus populations . However, among the sites in each province, there were few significant differences in the genotype compositions of virus populations . The number of genotypes detected at a site varied from two to nine with a few genotypes dominating . In general the isolates at a site persisted from season to season indicating a genetic stability for the local virus population . Over the sampling time, IRRI rice varieties, which have green leafhopper resistance genes, supported similar virus populations to those supported by other varieties, indicating that the variety of the host exerted no apparent selection pressures . Insect transmission experiments on selected RTBV field isolates showed that dramatic shifts in genotype and phenotype distributions can occur in response to host/environmental shifts. Cancer Immunol Immunother, 2000 Mar, 48(12), 703 - 13 Observed localization of the long-term cultured rat adherent natural killer cells in mammary tumor tissues; Gu S et al.; Adherent natural killer (A-NK) cells were isolated from splenic lymphocytes and treated with long-term culture in the presence of recombinant interleukin-2 (rIL-2) . Immunocytochemical and flow-cytometric analysis revealed that most of the A-NK cells strongly expressed lymphocyte-function-associated antigen 1 (LFA-1alpha, and LFA-1beta) throughout the incubation . All A-NK cells from 8- to 150-day cultures, particularly those cultured for 8 days, showed significant cytolytic activity against all targets . Analysis of the tissue distribution of the injected {3H} uridine-labeled A-NK cells demonstrated that, in the first 3 h, most (over 60%) cells localized in the lungs, and that most cells remained temporally within the cavities of blood capillaries of the lungs and moved gradually into lymphoid and other tissues . Peritumoral injection of various kinds of adjuvant, particularly Freund's complete adjuvant (FCA) plus bacillus Calmetee-Guerin (BCG), resulted in a marked accumulation of {3H}A-NK cells in mammary tumor tissues 24 h after injection, and simultaneously in the formation of vessels resembling high-endothelium venules (HEV), infiltration of LFA-1+ lymphocytes and expression of the ICAM-1 molecule on the tumor cells in the sites of tumor tissues . When 30 x 10(6) A-NK cells were intravenously administered, significant retardation of tumor growth and prolongation of survival of tumor-bearing rats were observed in the groups that received the prior injection of adjuvants, especially FCA + BCG and Freund's incomplete adjuvant (FIA) + BCG . The suppressive effect of combination therapy on tumor growth was blocked effectively by the injection of anti-ICAM-1 mAb . These results indicate that the prior injection of proper adjuvant into the peritumoral region is effective for the selective accumulation or infiltration of A-NK cells into the sites of tumor tissues, and results in the marked retardation of tumor growth. Arq Neuropsiquiatr, 1999 Sep, 57(3B), 723 - 34 Muscle involvement in leprosy . Study of the anterior tibial muscle in 40 patients; Werneck LC et al.; The involvement of skeletal striated muscle in leprosy is considered secondary due to peripheral neuropathy, but some studies point it to a primary muscle lesion . In order to investigate the muscle involvement in leprosy, we studied 40 patients (lepromatous 23, tuberculoid 13, borderline 2 and indeterminate 2) . The motor nerve conduction of the peroneal nerves had a reduction of the velocity, decreased compound muscle action potential and sometimes absence of potentials . The electromyographic study of the anterior tibial muscle showed signs of recent and chronic denervation in 77.5% of the cases and no myopathic potentials . The anterior tibial muscle biopsy revealed denervation in 45% of the cases, interstitial inflammatory myopathy in 30% and mixed (myopathic and neuropathic) pattern in 12.5% . Acid fast bacillus was detected in 25% of the cases, always in the interstitial tissue . Inflammatory reaction was present in the interstitial space and in patients with the lepromatous type . The histological findings clearly defined the presence of the so-called "Leprous Interstitial Myositis" on the top of denervation signs. J Urol, 2000 May, 163(5), 1588 - 90 Induction of mycobacteremia by intravesical bacillus Calmette-Guerin instillation in an experimental animal model and detection with polymerase chain reaction; Aygun C et al.; PURPOSE: The aim of this study was to detect mycobacteremia by polymerase chain reaction (PCR), induced by the instillation of bacillus Calmette-Guerin (BCG) to guinea pig bladder . We also investigated the peak time and the effect of the dose of BCG in injured and non-injured bladder . The sensitivities of routine culture and PCR were also compared . MATERIALS AND METHODS: Five different doses (0, 0.069, 0.69, 6.9 and 69 mg.) of BCG were instilled into 5 injured and 5 non-injured bladders . Blood samples were collected at 0, 5, 15, 30 and 60 minutes following instillation for routine culture and PCR for each dose . A total of 50 female guinea pigs were used . RESULTS: Three of 5 samples (60%) obtained 30 minutes after the instillation of 69 mg . BCG into injured bladders were PCR positive . Furthermore, 4 of 5 samples (80%) were PCR positive when samples were obtained at the 60th minute following instillation . All the other samples were negative for PCR and routine culture . All the routine tuberculosis culture results were negative, including those which were PCR positive . CONCLUSIONS: Mycobacteremia was detected only in injured bladders and with high doses of BCG . PCR is a highly sensitive and rapid diagnostic method for detection of mycobacteremia. J Urol, 2000 May, 163(5), 1553 - 9 Autocrine IL-6 production by human transitional carcinoma cells upregulates expression of the alpha5beta1 firbonectin receptor; Zhang GJ et al.; PURPOSE: Studies have demonstrated elevated expression and secretion of IL-6 by transitional cell