Can Respir J, 2000 Mar-Apr, 7(2), 141 - 6 Tuberculosis in aboriginal Canadians; Hoeppner VH et al.; Endemic tuberculosis (TB) was almost certainly present in Canadian aboriginal people (aboriginal Canadians denotes status Indians, Inuit, nonstatus Indians and metis as reported by Statistics Canada) before the Old World traders arrived . However, the social changes that resulted from contact with these traders created the conditions that converted endemic TB into epidemic TB . The incidence of TB varied inversely with the time interval from this cultural collision, which began on the east coast in the 16th century and ended in the Northern Territories in the 20th century . This relatively recent epidemic explains why the disease is more frequent in aboriginal children than in Canadian-born nonaboriginal people . Treatment plans must account for the socioeconomic conditions and cultural characteristics of the aboriginal people, especially healing models and language . Prevention includes bacillus Calmette-Guerin vaccination and chemoprophylaxis, and must account for community conditions, such as rates of suicide, which have exceeded the rate of TB . The control of TB requires a centralized program with specifically directed funding . It must include a program that works in partnership with aboriginal communities. Infect Immun, 2000 Jul, 68(7), 4084 - 91 High intracellular level of guanosine tetraphosphate in Mycobacterium smegmatis changes the morphology of the bacterium; Ojha AK et al.; Almost one-third of the world population today harbors the tubercle bacillus asymptomatically . It is postulated that the morphology and staining pattern of the long-term persistors are different from those of actively growing culture . Interestingly, it has been found that the morphology and staining pattern of the starved in vitro population of mycobacteria is similar to the persistors obtained from the lung lesions . In order to delineate the biochemical characteristics of starved mycobacteria, Mycobacteria smegmatis was grown in 0.2% glucose as a sole carbon source along with an enriched culture in 2% glucose . Accumulation of the stringent factor guanosine tetraphosphate (ppGpp) with a concomitant change in morphology was observed for M . smegmatis under carbon-deprived conditions . In addition, M . smegmatis assumed a coccoid morphology when ppGpp was ectopically produced by overexpressing Escherichia coli relA, even in an enriched medium . The Mycobacterium tuberculosis relA and spoT homologue, when induced in M . smegmatis, also resulted in the overproduction of ppGpp with a change in the bacterium's growth characteristics. Int J Food Microbiol, 2000 May 25, 56(1), 81 - 6 Potentiation of the lethal effect of peroxygen on Bacillus cereus spores by alkali and enzyme wash; Langsrud S et al.; Bacillus cereus present in pipes and heat-exchangers represents a potential quality problem for dairy industry . The peroxygen-containing disinfectants investigated had only negligible sporicidal effect when applied at the recommended in-use temperature and concentration . However, cleaning agents used before disinfection potentiated their lethal activity . Pre-exposure of B . cereus spores to 1% sodium hydroxide at temperatures over 40 degrees C increased the sporicidal effect of the peroxygen-containing disinfectant . The effect was dependent on the alkali concentration and the temperature . Also, a significant potentiating activity of an enzyme-based cleaning agent was obtained, but the effect was smaller than for alkali treatment . The results indicated that disinfectants based on peroxygen can be used to eliminate B . cereus spores at non-corrosive temperatures and concentrations if the surfaces are cleaned with alkali or enzyme-based disinfectants prior to disinfection. Ophthalmology, 2000 Jun, 107(6), 1038 - 41 Epidemic Bacillus endophthalmitis after cataract surgery II: chronic and recurrent presentation and outcome; Chen JC et al.; OBJECTIVE: To report the clinical outcome of chronic Bacillus endophthalmitis after cataract surgery . DESIGN: Retrospective, noncomparative case series . PARTICIPANTS: Five eyes of five patients with late-onset or recurrent inflammation after exposure to bacteria-contaminated viscoelastic material were studied . INTERVENTION: Repeated vitrectomies, wide excision of the remnant posterior capsule, and intravitreal injections of antibiotics in five patients . Eventual explantation of the intraocular lens in four patients . MAIN OUTCOME MEASURES: Final visual acuities and results of microbiologic studies of aqueous and vitreous specimens as well as pathologic studies using hematoxylin-eosin, Gram, and periodic acid-Schiff (PAS) stain of explanted capsular remnants were obtained . RESULTS: Final visual acuity of 20/40 or better was obtained in three patients . Bacillus species were grown from two cases . PAS- and Gram-positive microorganisms were identified in the capsular tissue in three of four patients who had explantation of the intraocular lens . CONCLUSIONS: A chronic form of Bacillus endophthalmitis is described for the first time . The clinical outcome is similar to chronic endophthalmitis caused by other organisms. Anal Chem, 2000 Jun 1, 72(11), 2428 - 32 Electron monochromator mass spectrometry for the analysis of whole bacteria and bacterial spores; Beverly MB et al.; Spores from a variety of Bacillus species were analyzed with direct probe mass spectrometry using an electron monochromator to select electrons of distinct energies for ionization . Electron energies were chosen to match the electron capture energies of taxonomically important compounds such as dipicolinic acid and fatty acids . Previous negative ion interferences were not observed when the monochromator was used, and the signal-to-noise ratio of targeted compounds was significantly enhanced using this approach . To demonstrate the selectivity of the technique, the monochromator was swept over a range of electron energies while monitoring the masses of compounds with known electron capture energies . Scanning the monochromator while the mass spectrometer was operated in single-ion mode enabled dipicolinic acid to be detected in 10(5) spores . The results presented here demonstrate the utility of the electron monochromator for selectively ionizing compounds directly in bacteria and bacterial spores. Comp Immunol Microbiol Infect Dis, 2000 Jul, 23(3), 153 - 62 Binding of bovine growth hormone to . Mycobacterium avium ss paratuberculosis; Feola RP et al.; Mycobacterium avium ss paratuberculosis causes a chronic progressive enteritis in cattle and other ruminants referred to as Johne's disease . It also has been suggested by some as possibly being associated with Crohn's disease in humans . In a previous study we observed that incubation of bovine monocytes with recombinant bovine growth hormone (bGH) altered the ingestion and intracellular growth of M . avium ss paratuberculosis in vitro . This led us to investigate whether bGH also has a direct effect on M . avium ss paratuberculosis . We observed that addition of bGH (5 microg/ml) had a direct inhibitory effect on the growth of M . avium ss paratuberculosis in Middlebrook 7H9 broth . In contrast, the growth of Mycobacterium smegmatis was unaffected, even at a bGH concentration of 50 microg/ml . Using 125I-bGH we observed high affinity binding (Kd = 1.32 nM) of bGH to M . avium ss paratuberculosis, with an estimated 204 binding sites per bacillus . To the best of our knowledge, this is the first report of a mammalian hormone binding to this important enteric pathogen. Graefes Arch Clin Exp Ophthalmol, 2000 Apr, 238(4), 299 - 301 Stenotrophomonas maltophilia endophthalmitis after intraocular lens implantation; Horio N et al.; BACKGROUND: Stenotrophomonas maltophilia is an opportunistic, gram-negative bacillus . Endophthalmitis induced by S . maltophilia has been described in only two cases after intraocular lens implantation . We report S . maltophilia endophthalmitis in two patients with diabetes mellitus after intraocular lens implantation and compare the characteristics of the S . maltophilia-induced endophthalmitis with two previous cases . METHODS: A 68-year-old woman and a 74-year-old man with diabetes mellitus developed S . maltophilia endophthalmitis within 5 days of intraocular lens implantation . We performed intraocular lens removal and vitrectomy, which resolved the inflammation . No recurrences were found . RESULTS: Cultures grew S . maltophilia in both cases, and one of the organisms was multi-resistant . The final visual acuity was counting fingers and 0.3 . The first case revealed a tractional retinal detachment during vitrectomy . CONCLUSIONS: S . maltophilia is a potential opportunistic intraocular pathogen, and the incidence of multiresistant S . maltophilia is increasing . S . maltophilia causes acute endophthalmitis, and its prognosis may not be poor unless the eye has a history of serious disease before the cataract surgery . The combined procedure of intraocular lens removal and vitrectomy was useful in resolving the inflammation and preventing recurrences. Oncology (Huntingt), 2000 May, 14(5), 719 - 29; discussion 729-31, 734, 737 Intravesical therapy for superficial bladder cancer; Baselli EC et al.; Approximately 54,400 new cases of transitional cell carcinoma of the bladder were reported in the United States in 1999, with an estimated 12,500 deaths attributable to this cancer . Close to 75% of all bladder tumors are confined to the urothelium (stage Ta, or carcinoma in situ), and nearly 30% of papillary tumors invade the lamina propria (stage T1) . The majority of superficial tumors are low grade with low rates of progression . Transurethral resection is the standard initial treatment for transitional cell carcinoma . Intravesical therapy is an important adjunct to transurethral resection in patients with superficial bladder cancer, many of whom are at risk for disease recurrence and progression . Cytotoxic and immunomodulating agents and, more recently, photosensitizers have demonstrated utility against superficial transitional cell carcinoma . Many studies have assessed and continue to examine the efficacy of various agents at different doses and in different combinations and schedules . Recently, valrubicin (Valstar) won Food and Drug Administration (FDA) approval only for the treatment of refractory carcinoma in situ . However, bacillus Calmette-Guerin (BCG) and mitomycin (Mutamycin) remain the most commonly used, most effective agents available for prophylaxis against recurrence and subsequent progression of superficial bladder cancer . This article reviews traditional and alternative intravesical agents useful in the therapy and prophylaxis of superficial transitional cell carcinoma of the bladder. Mol Gen Genet, 2000 May, 263(4), 571 - 80 A novel secreted ribonuclease from Bacillus intermedius: gene structure and regulatory control; Hahnen E et al.; A second secreted ribonuclease, designated binase II, has been detected in Bacillus intermedius 7P, and its structural gene was cloned and sequenced . Unlike the well-known binase I, a 109-amino acid guanyl-specific enzyme, the 292-residue binase II is closely related to the B . subtilis nuclease Bsn, in structure and in its enzymatic properties . Binase II is also insensitive to inactivation by barstar, an inhibitor protein that is specific for guanyl-specific ribonucleases . While both B . intermedius enzymes are induced upon phosphate starvation, only the gene for binase I belongs to the pho regulon system and carries pho-box elements adjacent to its promoter sequence . The gene for binase II is similar to that for Bsn in lacking such elements . The birB gene coding for binase II appears to be located next to the 3'-end of a ferric ion transport operon, with which it convergently overlaps . This would allow attenuator control over binase II expression under conditions of starvation for ferric ions. Mutat Res, 2000 May 31, 459(4), 307 - 16 Purification and characterization of a novel UV lesion-specific DNA glycosylase/AP lyase from Bacillus sphaericus; Vasquez DA et al.; The purification and characterization of a pyrimidine dimer-specific glycosylase/AP lyase from Bacillus sphaericus (Bsp-pdg) are reported . Bsp-pdg is highly specific for DNA containing the cis-syn cyclobutane pyrimidine dimer, displaying no detectable activity on oligonucleotides with trans-syn I, trans-syn II, (6-4), or Dewar photoproducts . Like other glycosylase/AP lyases that sequentially cleave the N--glycosyl bond of the 5' pyrimidine of a cyclobutane pyrimidine dimer, and the phosphodiester backbone, this enzyme appears to utilize a primary amine as the attacking nucleophile . The formation of a covalent enzyme-DNA imino intermediate is evidenced by the ability to trap this protein-DNA complex by reduction with sodium borohydride . Also consistent with its AP lyase activity, Bsp-pdg was shown to incise an AP site-containing oligonucleotide, yielding beta- and delta-elimination products . N-terminal amino acid sequence analysis of this 26 kDa protein revealed little amino acid homology to any previously reported protein . This is the first report of a glycosylase/AP lyase enzyme from Bacillus sphaericus that is specific for cis-syn pyrimidine dimers. J Invertebr Pathol, 2000 May, 75(4), 288 - 91 Effect of Bacillus thuringiensis toxins on the midgut of the nun moth Lymantria monacha; Rausell C et al.; Three steps of the proposed mode of action of Bacillus thuringiensis toxins have been studied in Lymantria monacha . We demonstrated that only the toxins that caused typical pathological changes in midgut epithelial cells and bound to the midgut brush border membrane were able to drastically reduce the midgut transepithelial voltage of the nun moth . J Invertebr Pathol, 2000 May, 75(4), 259 - 66 The interactions between soybean trypsin inhibitor and delta-endotoxin of Bacillus thuringiensis in Helicoverpa armigera larva; Zhang JH et al.; No significant difference in larval mortality was observed when a sublethal dose of Bacillus thuringiensis (Bt) var . kurstaki HD-1 crystal was supplemented with soybean trypsin inhibitor (STI) in the artificial diet fed to Helicoverpa armigera in the laboratory, but supplementing a nonlethal dose of crystal with STI in the diet led to a pronounced reduction of larval growth . This concentration of crystal and two lower concentrations of STI alone had no significant effects on larval growth . The results of substrate-gel electrophoresis demonstrated that the proteases in the H . armigera midgut fluid responsible for the degradation of protoxin consisted of at least four proteases with molecular weights of 71, 49, 36, and 30 kDa . All four proteases could utilize casein also as the substrate . When larvae were fed with STI or Bt + STI, the proteolytic activities of the 49-kDa enzyme disappeared, and the activities of the other three enzymes were reduced . Enzyme assays also indicated that feeding larvae with diets containing Bt, STI, or Bt + STI significantly decreased the specific activities of larval general proteases and the trypsin-like enzyme . The protein concentration of midgut fluid was elevated, especially in the larvae fed on the diets containing STI and Bt + STI . Both in vitro and in vivo studies showed that the degradation of protoxin and toxin could be inhibited by soybean trypsin inhibitors, but when the incubation time was prolonged, the protoxin could be degraded completely, while the degradation of toxin was inhibited further . This suggested that the retention time of toxins in the larval midgut was extended and synergism between insecticidal crystal protein and soybean trypsin inhibitor occurred, which showed as the inhibition of H . armigera larval growth . J Invertebr Pathol, 2000 May, 75(4), 243 - 50 Sedimentation of Bacillus sphaericus in tap water and sewage water; Skovmand O et al.; Formulations of Bacillus sphaericus products were applied in 1-m columns of clean water and sewage water in tubes . Samples taken through valves attached to the walls of the tubes showed that spores settled as a function of application method, droplet size, particle size, and water characteristics . In a mixture of deionized and tap water, the slowest sedimentation rate was obtained with a primary powder sonicated in water with 0.1% detergent . This product and a commercial fluid product settled faster in sewage water than in clean water and after 4 weeks, most spores were found in the sediment . In clean water, most spores remained suspended during the 4 weeks but the density was highest at the bottom . Calculation of total spore numbers in the columns after 4 weeks indicated that about 90% of spores disappeared in clean water, and 95-97% disappeared in sewage water . Bioassays showed that toxicity declined in parallel with the sedimentation, but at a higher rate, indicating a loss of toxicity from the residual spores . Appl Biochem Biotechnol, 2000 Spring, 84-86, 955 - 62 Enhancement of selectivity for producing gamma-cyclodextrin; Matioli G et al.; The production of cyclodextrins (CDs) by cyclodextrin-glycosyl-transferase (CGTase) from Bacillus firmus was studied, with respect to the effect of the source of starch upon CD yield and on the selectivity for producing gamma-CD . Cyclodextrin production tests were run for 24 h at 50 degrees C, pH 8.0, and 1 mg/L of CGTase, and substrates were maltodextrin or the starches of rice, potato, cassava, and corn hydrolyzed up to D.E . 10 . Cornstarch was the best substrate for producing gamma-CD . Later, glycyrrhizin (2.5% {w/v}), which forms a stable complex with gamma-CD, was added to the cornstarch reaction medium and increased the yield of gamma-CD to about four times that produced with only maltodextrin, but the total yield of CDs remained practically unchanged . Therefore, the results showed that the studied CGTase is capable of giving relatively high yield of gamma-CD in the presence of glycyrrhizin as complexant and cornstarch as substrate. Appl Biochem Biotechnol, 2000 Spring, 84-86, 903 - 15 Cloning and expression of the limonene hydroxylase of Bacillus stearothermophilus BR388 and utilization in two-phase limonene conversions; Cheong TK et al.; A 3.6-kb fragment of Bacillus stearothermophilus BR388 chromosomal DNA that confers growth on limonene to Escherichia coli has been sequenced, revealing a single open reading frame encoding a single subunit limonene hydroxylase containing 444 amino acid residues . This enzyme proved capable of limonene hydroxylation to a mixture of carveol and perillyl alcohol as well as dehydrogenation of these products to carvone and perillyl aldehyde . Oxygen, FAD, and NADH were found to stimulate the hydroxylation reaction in cell extracts, and NAD+ stimulated the dehydrogenase reaction . In two-phase bioconversions using viable E . coli cells over-expressing the limonene hydroxylase, perillyl alcohol and carvone were the principal products observed. Appl Biochem Biotechnol, 2000 Spring, 84-86, 655 - 63 Study of different media for production of penicillin G acylase from Bacillus megaterium ATCC 14945; Pinotti LM et al.; In this study, several fermentation media were tested for the production of penicillin G acylase (PGA) using Bacillus megaterium . The carbon sources studied were glucose and lactose . The nitrogen sources studied were enzymatic casein hydrolysates produced with proteases of different specificities . The replacement of glucose with cheese whey and the addition of free amino acids in the PGA production were also tested . The results showed a strong correlation between the nitrogen source and enzyme yield and the presence of glucose repression . The highest enzyme concentration achieved was 138 IU/L using casein hydrolyzed with 0.6 L of Alcalase and cheese whey. Eur J Biochem, 2000 Jun, 267(12), 3836 - 47 Export and folding of signal-sequenceless Bacillus licheniformis beta-lactamase in Escherichia coli; Frate MC et al.; Two genetically engineered variants of the Bacillus licheniformis beta-lactamase gene were expressed in Escherichia coli . One variant coded for the exo-small mature enzyme without the signal peptide . The other coded for the exo-large mature enzyme preceded by 10, mostly polar, residues from an incomplete heterologous signal . As observed following the extraction by a lysozyme-EDTA treatment, the signal-less variant was exported to the periplasm with nearly 20% efficiency, whereas the variant with the N-terminal extension was translocated to a lesser degree; interestingly, nearly all of the former and half of the latter were extracted by osmotic shock, which may be of importance for our understanding of cellular compartments . The fact that a signal-less protein is translocated with substantial yields raises questions about the essential role of signal peptides for protein export . As folding and export are related processes, we investigated the folding in vitro of the two variants . No differences were found between them . In the absence of denaturant, they are completely folded, fully active and have a large DeltaG of unfolding . Under partially denaturing conditions they populate several partially folded states . The absence of significant amounts of a non-native state under native conditions makes a thermodynamic partitioning between folding and export less likely . In addition, kinetic measurements indicated that these B . licheniformis lactamases fold much faster than E . coli beta-lactamase . This behavior suggests that they are exported by a kinetically controlled process, mediated by one or more still unidentified interactions that slow folding and allow a folding intermediate to enter the export pathway. Eur J Biochem, 2000 Jun, 267(12), 3432 - 41 The role of arginine 47 in the cyclization and coupling reactions of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 implications for product inhibition and product specificity; van der Veen BA et al.; Cyclodextrin glycosyltransferase (CGTase) (EC 2.4.1.19) is used for the industrial production of cyclodextrins . Its application, however, is hampered by the limited cyclodextrin product specificity and the strong inhibitory effect of cyclodextrins on CGTase activity . Recent structural studies have identified Arg47 in the Bacillus circulans strain 251 CGTase as an active-site residue interacting with cyclodextrins, but not with linear oligosaccharides . Arg47 thus may specifically affect CGTase reactions with cyclic substrates or products . Here we show that mutations in Arg47 (to Leu or Gln) indeed have a negative effect on the cyclization and coupling activities; Arg47 specifically stabilizes the oligosaccharide chain in the transition state for these reactions . As a result, the mutant proteins display a shift in product specificity towards formation of larger cyclodextrins . As expected, both mutants also showed lower affinities for cyclodextrins in the coupling reaction, and a reduced competitive (product) inhibition of the disproportionation reaction by cyclodextrins . Both mutants also provide valuable information about the processes taking place during cyclodextrin production assays . Mutant Arg47-->Leu displayed an increased hydrolyzing activity, causing accumulation of increasing amounts of short oligosaccharides in the reaction mixture, which resulted in lower final amounts of cyclodextrins produced from starch . Interestingly, mutant Arg47-->Gln displayed an increased ratio of cyclization/coupling and a decreased hydrolyzing activity . Due to the decreased coupling activity, which especially affects the production of larger cyclodextrins, this CGTase variant produced the various cyclodextrins in a stable ratio in time . This feature is very promising for the industrial application of CGTase enzymes with improved product specificity. Microbiology, 2000 Jun, 146 ( Pt 6), 1371 - 80 Tripartite haemolysin BL: isolation and characterization of two distinct homologous sets of components from a single Bacillus cereus isolate; Beecher DJ et al.; Haemolysin BL (HBL), a three-component enterotoxic/necrotizing/vascular permeability toxin, is a likely virulence factor of Bacillus cereus diarrhoeal food poisoning and necrotic infections . This paper describes the isolation of two distinct homologous sets of all three HBL components from a single B . cereus isolate, MGBC 145 . The proteins of one set (designated HBL, consisting of B, L(1) and L(2)), were about 87-100% identical in N-terminal amino acid sequences to their respective prototype components from strain F837/76, and the proteins of the homologous set (HBL(a), consisting of B(a), L(1a) and L(2a)) were all about 62-65% identical . Only the latter homologues differed immunochemically and physicochemically from the prototypes . HBL and HBL(a) exhibited similar haemolytic and vascular permeability potencies, and the homologues could be interchanged freely . There were no notable differences in activity between the L component homologues . However, components B and B(a) were significantly different . Both were secreted as monomers, but unlike B, B(a) was isolated as a relatively inactive complex that could be reactivated with urea . When B(a) was substituted for B in gel-diffusion assays the distinct discontinuous haemolysis pattern typical of the presence of B did not occur . In suspension assays, excess B inhibited the haemolysis of B-primed cells by L(1) (as previously described), but not that of B(a)-primed cells . Excess B(a) had the opposite effect and enhanced lysis of B(a)-primed cells, but not that of B-primed cells . These differences reveal details about how the toxin components interact on target cell membranes . The authors' observations indicate that HBL represents a new family of multicomponent toxins that was generated by a process of gene and operon duplication that occurred either intracellularly or by horizontal transfer, and raise the possibility of the existence of other related toxins in the genetically diverse B . cereus taxonomic group. Clin Exp Immunol, 2000 Jun, 120(3), 476 - 82 Protective effect of DNA immunization against mycobacterial infection is associated with the early emergence of interferon-gamma (IFN-gamma)-secreting lymphocytes; Kamath AT et al.; The development of more effective anti-tuberculosis (TB) vaccines would contribute to the global control of TB . Understanding the activated/memory T cell response to mycobacterial infection and identifying immunological correlates of protective immunity will facilitate the design and assessment of new candidate vaccines . Therefore, we investigated the kinetics of the CD4+ T cell response and IFN-gamma production in an intravenous challenge model of Mycobacterium bovis bacille Calmette-Guerin (BCG) before and after DNA immunization . Activated/memory CD4+ T cells, defined as CD44hiCD45RBlo, expanded following infection, peaking at 3-4 weeks, and decreased as the bacterial load fell . Activated/memory CD4+ T cells were the major source of IFN-gamma and the level of antigen-specific IFN-gamma-secreting lymphocytes, detected by ELISPOT, paralleled the changes in bacterial load . To examine the effects of a DNA vaccine, we immunized mice with a plasmid expressing the mycobacterial secreted antigen 85B (Ag85B) . This led to a significant reduction in mycobacteria in the liver, spleen and lung . This protective effect was associated with the rapid emergence of antigen-specific IFN-gamma-secreting lymphocytes which were detected earlier, at day 4, and at higher levels than in infected animals immunized with a control vector . This early and increased response of IFN-gamma-secreting T cells may serve as a correlate of protective immunity for anti-TB vaccines. Int J Urol, 2000 Jun, 7(6), 206 - 9 Clinical experience with BCG alone versus BCG plus epirubicin; Bilen CY et al.; BACKGROUND: Bacillus Calmette-Guerin (BCG) and epirubicin have both been shown to be effective in the treatment of superficial bladder cancer . We studied whether the alternating combination of these agents could improve the efficacy with tolerable side-effects in the treatment of high-risk superficial bladder tumors . METHODS: Forty-one patients with high-risk superficial bladder transitional carcinoma were included in this study . Twenty-one patients were randomized into the BCG group and 20 patients were treated with sequential BCG and epirubicin . The patients were followed for 9-24 months (mean 18 months) . Recurrence rates, median time to the first recurrence, progression rate and complications were compared . RESULTS: Fifteen percent of the patients in the BCG and epirubicin group and 19% of the patients in the BCG alone group developed tumor recurrence . Tumor progression was observed in 4.7% and 10% in the BCG/epirubicin group and the BCG alone group, respectively . Median time to first recurrence was 11 months for the BCG/epirubicin group and 16 months for the BCG group (P > 0.05) . Three patients in the BCG/epirubicin treatment group developed serious side-effects, which necessitated antituberculosis treatment . CONCLUSION: Because the efficacy of combination was no better than the standard treatment and the alternating combination seemed to be related to a higher incidence of side-effects, this study albeit small, does not recommend combination therapy of BCG and epirubicin in high risk patients with superficial bladder cancer. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 1305 - 13 New psychrophilic and psychrotolerant Bacillus marinus strains from tropical and polar deep-sea sediments and emended description of the species; Ruger HJ et al.; In contrast to the current view that psychrophily combined with an absolute requirement for NaCl is connected with the Gram-negative cell wall type, psychrophilic and psychrotolerant, NaCl-requiring, Gram-positive bacteria have been isolated from tropical Atlantic, Arctic and Antarctic deep-sea sediments . Some of the isolates are even extremely psychrophilic, having maximum growth temperatures of 4 degrees C . On the basis of phenotypic characteristics, DNA base analyses, DNA-DNA hybridizations and partial and complete 16S rRNA gene sequence analyses, the strains from the three distinct geographical regions have been allocated to the obligately marine species Bacillus marinus . The distribution and origin of B . marinus are discussed and an emended description of the species is presented. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 1081 - 6 Phylogenetic analysis of the genus Thermoactinomyces based on 16S rDNA sequences; Yoon JH et al.; The almost complete 16S rDNA sequences of the type strains of eight validly described species and two invalid species of the genus Thermoactinomyces were determined and phylogenetically analysed . The validly described Thermoactinomyces species formed phylogenetic lineages related to the family Bacillaceae, as shown previously . However, the available strains of 'Thermoactinomyces glaucus' and 'Thermoactinomyces monosporus' exhibited their closest phylogenetic affinities not to the genus Thermoactinomyces but to the genus Saccharomonospora . Some Thermoactinomyces species were shown to be closely related from 16S rDNA sequence analysis . Particularly, Thermoactinomyces vulgaris KCTC 9076T and Thermoactinomyces candidus KCTC 9557T had the same 16S rDNA sequences and Thermoactinomyces thalpophilus KCTC 9789T and Thermoactinomyces sacchari KCTC 9790T showed 16S rDNA similarity value of almost 100% . From phylogenetic analysis based on 16S rDNA sequences, it is suggested that the genus Thermoactinomyces should be taxonomically re-evaluated using other useful taxonomic markers. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 1021 - 8 Protein-coding genes as molecular markers for ecologically distinct populations: the case of two Bacillus species; Palys T et al.; Bacillus globisporus and Bacillus psychrophilus are one among many pairs of ecologically distinct taxa that are distinguished by very few nucleotide differences in 16S rRNA gene sequence . This study has investigated whether the lack of divergence in 16S rRNA between such species stems from the unusually slow rate of evolution of this molecule, or whether other factors might be preventing neutral sequence divergence at 16S rRNA as well as every other gene . B . globisporus and B . psychrophilus were each surveyed for restriction-site variation in two protein-coding genes . These species were easily distinguished as separate DNA sequence clusters for each gene . The limited ability of 16S rRNA to distinguish these species is therefore a consequence of the extremely slow rate of 16S rRNA evolution . The present results, and previous results involving two Mycobacterium species, demonstrate that there exist closely related species which have diverged long enough to have formed clearly separate sequence clusters for protein-coding genes, but not for 16S rRNA . These results support an earlier argument that sequence clustering in protein-coding genes could be a primary criterion for discovering and identifying ecologically distinct groups, and classifying them as separate species. J Urol, 2000 Jul, 164(1), 47 - 52 Weekly mitomycin C followed by monthly bacillus Calmette-Guerin or alternating monthly interferon-alpha2B and bacillus Calmette-Guerin for prophylaxis of recurrent papillary superficial bladder carcinoma; Kaasinen E et al.; PURPOSE: We evaluated alternatives to bacillus Calmette-Guerin (BCG) monotherapy using a new combination of chemotherapy and immunotherapy for recurrent superficial bladder carcinoma . MATERIALS AND METHODS: A total of 236 patients with frequently recurrent stage Ta or T1 bladder tumors were enrolled in our prospective, randomized, multicenter Finnbladder IV study . The initial mitomycin C instillation was instilled in all patients perioperatively after transurethral resection, followed by 4 weekly instillations of mitomycin C . Thereafter patients were randomized to receive monthly for up to 1 year BCG only or interferon-alpha2b and BCG alternating monthly . Primary end points were time to initial recurrence, recurrence rate (number of recurrences per patient-year) and recurrence index (number of recurrent tumors per patient-year) . RESULTS: Of the 236 randomized patients 205 were eligible for study with a median overall followup of 30.7 months . Monthly BCG was superior to alternating monthly interferon-alpha and/or BCG with respect to time to initial recurrence (log rank test p <0.00001) as well as recurrence rate (0.4 versus 0.9, p <0.00001) and index (0.9 versus 3.0, p <0.00001) . Side effects were limited . CONCLUSIONS: Monthly BCG given for up to 1 year preceded by perioperative and an additional 4 weekly mitomycin C instillations is a well tolerated mode of instillation therapy, providing excellent tumor control comparable to that of the best reported instillation regimens . No benefit was obtained by alternating interferon-alpha2b with BCG. Proc Natl Acad Sci U S A, 2000 Jul 5, 97(14), 7700 - 3 Absence of toxicity of Bacillus thuringiensis pollen to black swallowtails under field conditions; Wraight CL et al.; A single laboratory study on monarch butterflies has prompted widespread concern that corn pollen, engineered to express Bacillus thuringiensis (Bt) endotoxin, might travel beyond corn fields and cause mortality in nontarget lepidopterans . Among the lepidopterans at high potential risk from this technology is the black swallowtail butterfly, Papilio polyxenes, whose host plants in the midwestern U . S . are located principally in narrow strips between roads and crop fields . A field study was performed to assess whether mortality of early instar black swallowtails was associated either with proximity to a field of Bt corn or by levels of Bt pollen deposition on host plants . Potted host plants were infested with first instar black swallowtails and placed at intervals from the edge of a field of Bt corn (Pioneer 34R07 containing Monsanto event 810) at the beginning of anthesis . We confirmed by ELISA that pollen from these plants contained Cry1Ab endotoxin (2.125 +/- 0.289 ng/g) . Although many of the larvae died during the 7 days that the experiments were run, there was no relationship between mortality and proximity to the field or pollen deposition on host plants . Moreover, pollen from these same plants failed to cause mortality in the laboratory at the highest pollen dose tested (10,000 grains/cm(2)), a level that far exceeded the highest pollen density observed in the field (200 grains/cm(2)) . We conclude that Bt pollen of the variety tested is unlikely to affect wild populations of black swallowtails . Thus, our results suggest that at least some potential nontarget effects of the use of transgenic plants may be manageable. Chemistry, 2000 May 2, 6(9), 1531 - 6 Directed evolution of the fatty-acid hydroxylase P450 BM-3 into an indole-hydroxylating catalyst; Li QS et al.; The self-sufficient cytochrome P450 BM-3 enzyme from Bacillus megaterium catalyzes subterminal hydroxylation of saturated long-chain fatty acids and structurally related compounds . Since the primary structure of P450 BM-3 is homologous to that of mammalian P450 type II, it represents an excellent model for this family of enzymes . During studies on the directed evolution of P450 BM-3 into a medium-chain fatty-acid hydroxylase, several mutants, in particular the triple mutant Phe87Val, Leu188Gln, Ala74Gly, were observed to hydroxylate indole, producing indigo and indirubin at a catalytic efficiency of 1365 M(-1)s(-1) (kcat=2.73 s(-1) and Km=2.0 mM) . Both products were unequivocally characterized by NMR and MS analysis . Wild-type P450 BM-3 is incapable to hydroxylate indole . These results demonstrate that an enzyme can be engineered to catalyze the transformation of substrates with structures widely divergent from those of its native substrate. Probl Tuberk, 2000, (2), 9 - 13 {Tuberculosis morbidity in the Sverdlovsk region}; Nechaeva OB et al.; Childhood and adolescence tuberculosis morbidity rates are higher in girls than in boys . Adult males fall ill 5.3 times more frequently than adult females . Males fall ill with destructive (6.1 times), bacillary (4.1 times), pulmonary tuberculosis (5.3 times) more frequent than females . Females experience extrapulmonary tuberculosis 1.6 times as high as males . Convicts and those under investigation are ascertained to have earlier forms of respiratory tuberculosis than in the general population . At the same time, the facilities of the RF Ministry of Justice do not record extrapulmonary tuberculosis . Goal-oriented work with these facilities has caused a reduction in tuberculosis morbidity in the whole Sverdlovsk region. Jpn J Cancer Res, 2000 May, 91(5), 534 - 42 Enhancing effect of an inhibitor of nitric oxide synthesis on bacillus Calmette-Guerin-induced macrophage cytotoxicity against murine bladder cancer cell line MBT-2 in vitro; Yamada H et al.; We studied the effect of an inhibitor of nitric oxide (NO) synthesis, N(G)-monomethyl-L-arginine (L-NMMA), on the Bacillus Calmette-Guerin (BCG)-induced antitumor activity of murine peritoneal exudate cells (PEC) against murine bladder cancer cell line MBT-2 in vitro . L-NMMA enhanced BCG-induced cytotoxic activity of PEC, as well as interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha production . The L-NMMA-induced enhancement was due to the prolonged survival of BCG in macrophages, because no enhancement of cytotoxicity was observed and neither IFN-gamma nor TNF-alpha production was significantly enhanced by killed BCG . Anti-TNF-alpha antibody (Ab) and anti-IFN-gammaAb reduced the L-NMMA-induced enhancement of the cytotoxicity . The depletion of T cells from PEC reduced the production of both IFN-gamma and TNF-alpha, as well as the enhancement of cytotoxicity induced by viable BCG plus L-NMMA . These results suggest that L-NMMA has an enhancing effect on BCG-induced macrophage cytotoxicity and the enhancement is partially mediated by T cells and their soluble products . Accordingly, NO inhibitor should be a valuable adjunct to BCG immunotherapy for bladder cancer. Int J Lepr Other Mycobact Dis, 2000 Mar, 68(1), 40 - 8 Interferon-gamma responses to candidate leprosy skin-test reagents detect exposure to leprosy in an endemic population; Manandhar R et al.; New tools for the detection of leprosy exposure in a community will be necessary for the eradication of leprosy . Candidate leprosy skin-test antigens derived from the fractionation of the leprosy bacillus into cytoplasmic and cell-wall proteins free of immuno-inhibitory mycobacterial lipoglycans and carbohydrates were used in an overnight blood test to determine whether exposure to leprosy can be detected by the production of the cytokine interferon gamma (IFN-gamma) . Strong IFN-gamma responses were detected in leprosy contacts to both skin-test antigens compared with control subjects from the same endemic communities . There was little response in patients with tuberculosis . Responses were greatest in contacts with recent leprosy exposure . The implications of these findings for the application of these reagents in a field trial as skin tests to detect exposure to leprosy are discussed in light of the strong association between overnight IFN-gamma to PPD and the tuberculin skin-test responses previously reported. Int J Lepr Other Mycobact Dis, 2000 Mar, 68(1), 27 - 39 Antileprosy protective vaccination of rhesus monkeys with BCG or BCG plus heat-killed Mycobacterium leprae: immunologic observations; Gormus BJ et al.; Groups of rhesus monkeys were vaccinated and boosted with Mycobacterium bovis bacillus Calmette Guerin (BCG) or BCG plus low-dose (LD) or high-dose (HD) heat-killed M . leprae (HKML), or were unvaccinated . Prior to and following vaccination-boosting and subsequent M . leprae (ML) challenge, these and unvaccinated, unchallenged control monkeys were observed longitudinally for approximately 3 years . Vaccination with BCG plus HKML initially stimulated significant in vitro blood mononuclear cell blastogenic responses to lepromin, which returned to baseline post-boosting and post-live-ML-challenge, minimally reappearing significantly 2 years post-ML-challenge . Vaccination with BCG failed to stimulated positive blastogenic responses to lepromin before ML-challenge but small, marginally positive, intermittent responses were seen post-ML-challenge . Compared to the unvaccinated ML-challenged group, significant increases in the numbers of blood CD4+ and CD8+ T-cell subsets and an increased CD4+:CD8+ ratio were observed in both BCG plus HKML-vaccinated, ML-challenged groups, but not in the BCG-only-vaccinated, ML-challenged group . CD4+CD29+ and CD4+CD45RA+ subset numbers increased significantly over time in only the BCG plus LD HKML-vaccinated, ML-challenged group . Compared to unvaccinated, ML-challenged groups, vaccination with BCG or BCG plus HKML followed by ML-challenge produced lower IgM:IgG antiphenolic glycolipid-I (PGL-I) serum antibody ratios and protected rhesus monkeys from clinical leprosy, consistent with prior observations that low IgM:IgG anti-PGL-I responses correlated with resistance to and protection from leprosy. Folia Parasitol (Praha), 2000, 47(1), 45 - 8 Bacillary band ultrastructure of the fish parasite Capillaria pterophylli (Nematoda: Capillariidae); Zd'arska Z et al.; In Capillaria pterophylli Heinze, 1933, two lateral bacillary bands extend along the whole body in female and male worms . A ventral bacillary band is present in females only . The bacillary bands consist of glandular and non-glandular cells, in the region between the nerve ring and the end of the stichosome, ciliated sense receptors in tight connection with gland are present. Indian J Pediatr, 1996 Jul-Aug, 63(4), 569 - 70 Septic arthritis due to Actinobacillus actinomycetemcomitans; Shenoy S et al.; Actinobacillus actinomycetemcomitans, a gram-negative aerobic bacillus of the family Parvobacteriaceae which is a normal inhabitant of the oral flora, is a rare cause of human infection . We report a case of septic arthritis caused by this organism in an uncompromised child. Appl Environ Microbiol, 2000 Jun, 66(6), 2631 - 5 Cloning, expression, and sequence analysis of the gene encoding the alkali-stable, thermostable endoxylanase from alkalophilic, mesophilic Bacillus sp . Strain NG-27; Gupta N et al.; Alkalophilic Bacillus sp . strain NG-27 produces a 42-kDa endoxylanase active at 70 degrees C and at a pH of 8.4 . The gene for this endoxylanase was cloned and sequenced . The gene contained one open reading frame of 1,215 bases . An active site characteristic of the family 10 beta-glycanases was recognized between amino acids 303 and 313, with the active glutamate at position 310 . Though highly thermostable, the enzyme contains no cysteine residue. Microbiol Res, 2000 Apr, 155(1), 23 - 9 Larval susceptibility of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae), to Bacillus thuringiensis H serovars isolated in Japan; Higuchi K et al.; A total of 1700 Japanese strains of Bacillus thuringiensis, belonging to at least 47 H serogroups, were examined for insecticidal activity against larvae of the diamondback moth, Plutella xylostella . The high-level toxicity was associated with 612 isolates (36.0%) . Of these, 608 isolates (99.3%) fell into 13 H serogroups belonging to the low-numbered H serotypes, H1-H10 . Conversely, most isolates belonging to the high-numbered serotypes (>H10) had little or no larvicidal activity; only one isolate of the serovar japonensis H23 was active . P xylostella larvae were susceptible to 89.8% of the serovar morrisoni H8a:8b strains and 85.7% of galleriae H5a:5b strains . High values of 60-80% were also obtained in six serovars (thuringiensis H1, alesti H3a:3c, kurstaki H3a:3b:3c, kenyae H4a:4c, aizawai H7, and tolworhi H9), while relatively low values of <60% in two other common serovars, sotto H4a:4b and darmstadiensis H10a:10b . Five selected isolates, belonging to H serovars other than kurstaki and aizawai, were 10-60 times less toxic than the reference strain HD-1 (serovar kurstaki) . Parasporal inclusion proteins of these strains were immunologically unrelated to those of the strain HD-1 and the aizawai type strain. Microbiol Res, 2000 Apr, 155(1), 17 - 22 Bacillus thuringiensis soil populations naturally occurring in the Ryukyus, a subtropic region of Japan; Ohba M et al.; Of 809 soil samples collected from the seven islands of the Ryukyus, Japan, 107 samples (13.2%) contained Bacillus thuringiensis . The frequency of B . thuringiensis among the B . cereus group was 1.1% (235/21842) on the average . The B . thuringiensis soil populations of the Ryukyus consisted of more than 22 H serogroups . The predominant H serotype was the H5ac/21 (serovar canadensis/colmeri), followed by the H3ad (serovar sumiyoshiensis) and H16 (serovar indiana) . Geographically, most widely distributed H serogroups were the H16 and H10ac (serovar londrina); the former was recovered from five islands and the latter from three islands . Parasporal inclusions of the isolates were morphologically heterogeneous, roughly grouped into four categories: bipyramidal/cuboidal, spherical/ovoid, irregularly-pointed, and irregular-shaped . About 53% of the isolates formed spherical to ovoid parasporal inclusions . None of the isolates exhibited larvicidal activity against the silkworm, Bombyx mori . Only four isolates belonging to four different serotypes killed larvae of the mosquito, Aedes aegypti . These mosquito-specific isolates all produced spherical parasporal inclusions. Biosci Biotechnol Biochem, 2000 Apr, 64(4), 887 - 90 Purification and partial characterization of a basic xylanase produced by thermoalkaliphilic Bacillus sp . strain TAR-1; Takahashi H et al.; A basic xylanase was purified from the culture supernatant of thermoalkaliphilic Bacillus sp . strain TAR-1 . Its molecular mass and isoelectric point were 23 kDa and > pH 9.3, respectively . The enzyme showed a broad pH profile and was optimally active at 70 degrees C . Analyses of xylan-degradation products and N-terminal amino acid sequence revealed that the enzyme would be a family 11/G endoxylanase. Biochemistry, 2000 May 30, 39(21), 6538 - 45 Enzyme-induced strain/distortion in the ground-state ES complex in beta-lactamase catalysis revealed by FTIR; Hokenson MJ et al.; Class A beta-lactamases hydrolyze penicillins and other beta-lactams via an acyl-enzyme catalytic mechanism . Ser70 is the active site nucleophile . By constructing the S70A mutant, which is unable to form the acyl-enzyme intermediate, it was possible to make stable ES complexes with various substrates . The stability of such Michaelis complexes permitted acquisition of their infrared spectra . Comparison of the beta-lactam carbonyl stretch frequency (nu(CO)) in the free and enzyme-bound substrate revealed an average decrease of 13 cm(-)(1), indicating substantial strain/distortion of the lactam carbonyl when bound in the ES complex . Interestingly, regardless of the frequency of the C=O stretch in the free substrate, when complexed to Bacillus licheniformis beta-lactamase, the frequency was always 1755 +/- 2 cm(-)(1) . This suggests the active site environment induces a similar conformation of the beta-lactam in all substrates when bound to the enzyme . Using deuterium substitution, it was shown that the "oxyanion hole", which involves hydrogen bonding to two backbone amides, is the major source of the enzyme-induced strain/distortion . The very weak catalytic activity of the S70A beta-lactamase suggests enzyme-facilitated hydrolysis due to substrate distortion on binding to the enzyme . Thus the binding of the substrate in the active site induces substantial strain and distortion that contribute significantly to the overall rate enhancement in beta-lactamase catalysis. Eur Urol, 2000, 37 Suppl 3, 50 - 3 Efficacy of prophylactic Immucothel in patients pretreated with conventional drugs to prevent recurrence of superficial bladder carcinoma; Echarti C et al.; OBJECTIVES: Fifty patients enrolled in clinical trials with Immucothel((R)) were reevaluated for their disease-free intervals . METHODS: Of the 37 evaluable patients, 16 had received mitomycin C, 3 bacillus Calmette-Guerin, 3 Adriamycin and 15 miscellaneous pretreatments prior to Immucothel . They thus served as their own controls . RESULTS: Although their prognosis was worse at the time when Immucothel therapy was started than at the time of initiation of pretreatment, the mean recurrence-free intervals of all patients increased from 17.0 months under pretreatment to 35.1 months under Immucothel . CONCLUSION: The difference was statistically significant (Wilcoxon matched-pairs signed-ranks test) . Eur Urol, 2000, 37 Suppl 3, 45 - 9 Keyhole limpet hemocyanin for carcinoma in situ of the bladder: a long-term follow-up study; Jurincic-Winkler CD et al.; OBJECTIVE: Keyhole limpet hemocyanin (KLH) is a nonspecific immunomodulator, demonstrated to be clinically effective in superficial bladder cancer . The present study investigated the clinical efficacy of intravesical KLH in patients with carcinoma in situ (CIS) with a long-term follow-up . METHODS: Thirteen patients with CIS grade III were treated with intravesical instillations of KLH, 20 mg for 6 weeks, then monthly for 1 year and bimonthly for 2 subsequent years . Patients not responding to 2 courses of KLH were treated with bacillus Calmete-Guerin (BCG, 81 mg Connaught strain) . RESULTS: The follow-up period ranged from 12 to 84 months . Two patients were free of tumor after KLH instillations with a follow-up of 66 and 82 months, respectively . All patients who did not respond to the primary KLH course, but to the 'rescue' instillation of BCG, experienced recurrences after 42, 48, 56 and 60 months after the first KLH instillation treatment . Three patients with recurrent CIS and who were not cystectomized had recurrences after prolonged remission (4-5 years) . Patients progressing despite KLH and BCG instillations underwent cystectomy . CONCLUSIONS: KLH demonstrates efficacy and induces long- term remissions against CIS in a limited number of cases . In the present study, most patients with CIS progressed over time whatever the substance instilled, whether KLH or BCG . CIS remains a very aggressive neoplasm requiring a lifelong follow-up . Further studies are necessary to define the precise role of KLH in patients with CIS . Eur Urol, 2000 Jun, 37(6), 654 - 9 Prognostic value of MIB-1 antibody labeling index to predict response to Bacillus Calmette-Guérin therapy in a high-risk selected population of patients with stage T1 grade G3 bladder cancer; Lebret T et al.; PURPOSE: Proliferation rate is an important determinant of bladder tumor progression . However, this factor has not yet been correlated to bacillus Calmette-Guerin (BCG) therapy response in a selected high-risk population of patients with stage T1 grade G3 bladder cancer . To assess the predictive value of the proliferation rate, an immunoreactivity test with monoclonal antibodies MIB-1 was carried out . The aim of this study was to evaluate the prognostic value of an MIB-1 labeling index by selecting a group of responsive patients prior to intravesical therapy . MATERIALS AND METHODS: After complete transurethral resection, 35 patients with T1G3 bladder carcinoma received 6 weekly installations of BCG (intravesical Pasteur strain: 75 mg in 50 ml course of BCG therapy) . After treatment a cystoscopy and randomized biopsies of the bladder mucosa were carried out and all recurrences were systematically resected . All tissue samples were fixed in Bouin's solution, embedded in paraffin and stained with hematoxylin-eosin-safran . Pathologists had sufficient material to perform immunomarking in 25 patients using peroxidase-antiperoxidase (PAP) technique, with antiprotein monoclonal antibody MIB-1 (Immunotech, Marseilles, France) to study MIB-1 expression before BCG therapy . Consensus was obtained from three independent pathologists for all sections . The results were expressed in a percentage of marked nuclei . Ten percent increment thresholds were established from 10 to 60% . Contingency tables were established, chi2 (p1) and Fisher exact test (p2) were performed for each threshold of 10% . RESULTS: Median follow-up was 57.3 months (range 25-144) . Of the 25 patients, 8 (32%) did not respond to BCG therapy, 17 (68%) responded positively . With a 20% threshold, there was a statistical difference (p1 = 0.03, p2 = 0.04) between responder (R) and nonresponder (NR) patients . All the 7 patients with less than 20% of nuclear activity positively responded to BCG . At this threshold level, sensitivity was high but specificity low (positive predictive value = 0.44) . If we consider other reactivity thresholds there were no statistical differences between R and NR patients (10%) threshold p1 = 0.13, p2 = 0.19; 30% p1 and p2 = 0.20; 40% p1 = 0.82, p2 = 0.61; 50% p1 = 0.57, p2 = 0.55) . CONCLUSION: Our study indicates that the proliferation rate, assessed by MIB-1 immunoreactivity in Bouin's solution-fixed primary tissue, could be a useful predictive marker of outcome in T1G3 bladder carcinoma . With a 20% reactivity cut-off, a negative MIB-1 immunomarking appears to predict a positive response to BCG instillations . However, on the other hand, MIB-1 is of limited clinical use because the low specificity of this test cannot predict failure and then select candidates for cystectomies. Curr Microbiol, 2000 Jun, 40(6), 402 - 8 Isolation and characterization of Bacillus thuringiensis strains from aquatic environments in Spain; Iriarte J et al.; Samples collected from aquatic environments from Spain were analyzed for the occurrence and dipteran toxicity of Bacillus thuringiensis . From a total of 41 samples, 122 isolates were obtained, yielding a B . thuringiensis index of 0.22 . Isolates were assigned to 13 different serovars, with serovar thuringiensis (serotype H1) the most frequently found . Toxicity tests carried out revealed that eight isolates (6.6% out of the total) were active against Tipula oleracea larvae . Serological tests assigned these toxic isolates to serovar thuringiensis . The toxicity found in these isolates against the tipulid was approximately seven times lower than that shown by the standard strain B . thuringiensis ser . israelensis IPS-82 . Implication of Cry2A protein in toxic activity is hypothesized. J Food Prot, 2000 May, 63(5), 620 - 4 Antimicrobial activity of carvacrol toward Bacillus cereus on rice; Ultee A et al.; The antimicrobial activity of carvacrol, a compound present in the essential oil fraction of oreganum and thyme, toward the foodborne pathogen Bacillus cereus on rice was studied . Carvacrol showed a dose-related inhibition of growth of the pathogen . Concentrations of 0.15 mg/g and higher inhibited the growth and the extent of inhibition depended on the initial inoculum size . To decrease the input of carvacrol on the taste and flavor of the product, a combined treatment with the structure analog cymene was tested . Due to the smell and taste of carvacrol at high concentrations, carvacrol was combined with cymene, a natural antimicrobial compound with a similar structure . A synergistic effect was observed when 0.30 mg/g carvacrol was combined with 0.27 mg/g cymene . Finally it was demonstrated that a common taste enhancer like soya sauce also increased the antimicrobial action of carvacrol toward B . cereus . The antimicrobial activity of carvacrol with cymene or soya sauce was influenced by the addition of NaCl. Biochim Biophys Acta, 2000 May 23, 1478(2), 221 - 31 Aggregation, dissociation and unfolding of glucose dehydrogenase during urea denaturation; Mendoza-Hernandez G et al.; The effect of urea on glucose dehydrogenase from Bacillus megaterium has been studied by following changes in enzymatic activity, conformation and state of aggregation . It was found that the denaturation process involves several transitions . At very low urea concentrations (below 0.5 M), where the enzyme is fully active and tetrameric, there is a conformational change as monitored by an increase in intensity of the tryptophan fluorescence and a maximum exposure of organized hydrophobic surfaces as reported by the fluorescence of 4,4'-dianilino-1,1'-binaphthyl-5.5'-disulfonic acid . At slightly higher urea concentrations (0.75-2 M), a major conformational transition occurs, as monitored by circular dichroism and fluorescence measurements, in which the enzyme activity is completely lost and is concomitant with the formation of interacting intermediates that lead to a highly aggregated state . Increasing urea concentrations cause a complete dissociation to lead first a partially and eventually the complete unfolded monomer . These phenomena are fully reversible by dilution of denaturant . It is concluded that after urea denaturation, the folding/assembly pathway of glucose dehydrogenase occurs with the formation of intermediate species in which transient higher aggregates appear to be involved. J Cell Physiol, 2000 Jul, 184(1), 27 - 36 Effects of pertussis toxin on extracellular signal-regulated kinase activation in hepatocytes by hormones and receptor-independent agents: evidence suggesting a stimulatory role of G(i) proteins at a level distal to receptor coupling; Melien O et al.; It was previously found that pertussis toxin (PTX) pretreatment inhibits the activation of extracellular signal-regulated kinases ERK1 (p44(mapk)) and ERK2 (p42(mapk)) in hepatocytes in response to either agonists that bind to heptahelical receptors or epidermal growth factor (EGF), suggesting a role of G(i) proteins in stimulatory mechanisms for ERK1/2 . The present work shows that ERK1/2 is activated in a PTX-sensitive way not only by vasopressin, angiotensin II, prostaglandin (PG) F(2alpha), alpha(1)-adrenergic stimulation, and EGF but also by agents whose actions bypass receptors and stimulate protein kinase C (PKC) and/or elevate intracellular Ca(2+), such as 12-O-tetradecanoyl phorbol-13-acetate (TPA), exogenous phosphatidylcholine-specific phospholipase C (PC-PLC, from Bacillus cereus), thapsigargin, and the Ca(2+) ionophore A23187 . Under the same conditions, PTX did not affect agonist stimulation of phosphoinositide-specific phospholipase C (PI-PLC) (IP(3) generation), and did not reduce the activation by these agents of phospholipase D (PLD) . The results suggest that in hepatocytes a PTX-sensitive mechanism, presumably involving G(i) proteins, exerts a stimulatory effect on ERK at a level distal to receptor coupling, acting either as an integral part of the signaling pathway(s) or by a permissive, synergistic regulation . Clin Infect Dis, 1999 Apr, 28(4), 785 - 90 Clinical reactogenicity of intradermal bacille Calmette-Guérin vaccination; Hoft DF et al.; Clinical, microbiological, and immunologic responses were evaluated in volunteers vaccinated intradermally with bacille Calmette-Guerin (BCG) . Most volunteers (98%) developed ulcerative lesions that drained for a mean +/- SE of 4.3 +/- 0.29 weeks . Mycobacterial DNA was detected by a polymerase chain reaction-based amplification technique in biopsy specimens from BCG ulcers 2 weeks after vaccination and in blood specimens 3 days after vaccination . Mycobacteria were cultured from ulcer drainage 2 months after vaccination, demonstrating a prolonged potential risk of contact spread of the vaccine strain . The duration of ulcer drainage was inversely correlated with prevaccination lymphoproliferative (r = -0.515; P < .002) and interferon gamma (r = -0.841; P < .002) responses specific to mycobacteria and directly correlated with postvaccination increases in lymphoproliferative (r = 0.498; P < .002) and interferon gamma (r = 0.688; P < .02) responses specific to mycobacteria . These results demonstrate the clinical reactogenicity of BCG and the potential risk of contact spread of the vaccine strain and suggest that clinical reactogenicity is a trade-off for the induction of protective mycobacterial immunity. J Econ Entomol, 2000 Apr, 93(2), 484 - 93 Effect of plant age, larval age, and fertilizer treatment on resistance of a cry1Ab-transformed aromatic rice to lepidopterous stem borers and foliage feeders; Alinia F et al.; The resistance of vegetative, booting, and flowering stage plants of a variety of an aromatic rice, Oryza sativa L., transformed with a Bacillus thuringiensis Berliner cry1Ab gene under control of the maize phosphoenolpyruvate carboxylase (PEPC) promoter was evaluated against four lepidopterous rice pests--the stem borers Chilo suppressalis (Walker) (Lepidoptera: Crambidae) and Scirpophaga incertulas (Walker) (Lepidoptera: Pyralidae), and the foliage feeders Cnaphalocrocis medinalis Guenee (Lepidoptera: Pyralidae) and Naranga aenescens Moore (Lepidoptera: Noctuidae) . Plants of the cry1Ab-transformed line (no . 827) were more resistant to young larvae of S . incertulas, C . suppressalis, and C . medinalis than control plants at the vegetative stage but not at the flowering stage . Survival of 10-d-old stem borer larvae did not differ on cry1Ab plants and control plants at either the vegetative or flowering stage, but the development of 10-d-old C . suppressalis larvae was retarded on the vegetative stage cry1Ab plants . Immunological analysis also showed an apparent decline in Cry1Ab titer in leaf blades and leaf sheaths at the reproductive stage . In experiments comparing three fertilizer treatments (NPK, PK, and none), there was a significant interaction between fertilizer treatment and variety on larval survival only in whole-plant assays at booting stage with C . suppressalis . On cry1Ab plants, larval survival did not differ significantly among the three fertilizer levels, whereas on control plants survival was highest with the NPK treatment . cry1Ab plants tested at the sixth and seventh generations after transformation were more resistant than control plants to N . aenescens and C . suppressalis, respectively, suggesting that gene silencing will not occur in line 827 . The results of the experiments are discussed in terms of resistance management for B . thuringiensis toxins in rice. J Econ Entomol, 2000 Apr, 93(2), 459 - 63 Susceptibility of Spanish populations of the corn borers Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) to a Bacillus thuringiensis endotoxin; Gonzalez-Nunez M et al.; Baseline susceptibility to the Cry1Ab delta-endotoxin from Bacillus thuringiensis (Berliner) was determined for four populations of Sesamia nonagrioides (Lefebvre) and two populations of Ostrinia nubilalis (Hubner) from Spain . This study shows that S . nonagrioides is at least as susceptible as O . nubilalis to B . thuringiensis Cry1Ab protein . We found small differences in susceptibility among the Spanish populations of S . nonagrioides that can be attributed to natural variation, because there are no records of B . thuringiensis products being used on corn crops in Spain . There were no differences in susceptibility to Cry1Ab toxin between the two populations of O . nubilalis. J Econ Entomol, 2000 Apr, 93(2), 219 - 25 Assessment of microencapsulated formulations for improved residual activity of Bacillus thuringiensis; Tamez-Guerra P et al.; Bacillus thuringiensis Berliner is a highly efficacious bioinsecticide used to control lepidopteran pests in the field . Unfortunately, it has limited residual activity on plants because sunlight inactivates spores and crystals and they can be washed off by rain . To minimize loss of activity, formulations must contain UV protectants, stickers, or both . We tested approximately 80 formulations and determined optimal combinations of ingredients and spray drying conditions for improving B . thuringiensis residual activity after simulated rain and simulated sunlight . B . thuringiensis stability, after simulated sunlight (xenon light/8 h) and rain (5 cm/50 min), was improved using formulations based on lignin, corn flours, or both, with up to 20% of the active ingredient, when compared with technical powder or Dipel 2x in laboratory assays . Two formulations, made with corn flours or lignin + pregelatinized corn flour (PCF), killed 51.6 and 75.3% of Ostrinia nubilalis (Hubner) neonates after rain, respectively, versus 27% for technical powder . When the insecticidal activity was tested after simulated sunlight, corn flour-based formulations killed 78.5% of test larvae, and the lignin + PCF formulation killed 70.4%, in contrast to technical powder which caused an average of 29% mortality . Formulations made with Dipel 2x rather than technical powder, caused 62.5% mortality (corn flour-based formulations), and 72.3% mortality (lignin + PCF), versus 53.4% for Dipel 2x after rain . When tested after simulated sunlight, formulations killed 95% of the larvae (average of both formulations) versus 82% for Dipel 2x . In a field test, formulations were applied to cabbage and insecticidal activity was determined against Trichoplusia ni (Hubner) neonates exposed to treated leaves . Insecticidal activity of the corn flour-based formulations was comparable to Dipel 2x for 4 d after treatment, but was significantly better than Dipel 2x 7 d after application . A lignin and PCF-based formulation showed significantly higher residual activity than Dipel 2x, 4 and 7 d after application. J Infect Dis, 2000 May, 181(5), 1850 - 4 Epub 2000 May 15. Detection of active tuberculosis infection by T cell responses to early-secreted antigenic target 6-kDa protein and culture filtrate protein 10; Arend SM et al.; The purified protein derivative (PPD) skin test has no predictive value for tuberculosis (TB) in Mycobacterium bovis bacillus Calmette-Guerin (BCG)-vaccinated individuals because of cross-reactive responses to nonspecific constituents of PPD . T cell responses to early-secreted antigenic target 6-kDa protein (ESAT-6) and the newly identified culture filtrate protein 10 (CFP-10), 2 proteins specifically expressed by M . tuberculosis (MTB) but not by BCG strains, were evaluated . Most TB patients responded to ESAT-6 (92%) or CFP-10 (89%) . A minority of BCG-vaccinated individuals responded to both ESAT-6 and CFP-10, their history being consistent with latent infection with MTB in the presence of protective immunity . No responses were found in PPD-negative controls . The sensitivity and specificity of the assay were 84% and 100%, respectively, at a cutoff of 300 pg of interferon-gamma/mL . These data indicate that ESAT-6 and CFP-10 are promising antigens for highly specific immunodiagnosis of TB, even in BCG-vaccinated individuals. J Infect Dis, 2000 May, 181(5), 1846 - 9 Epub 2000 May 09. CD4+ and CD8+ T cells mediate adoptive immunity to aerosol infection of Mycobacterium bovis bacillus Calmette-Gu¿erin; Feng CG et al.; An adoptive-transfer model using recombinase activation gene-deficient (RAG-1-/-) mice was developed to evaluate CD4+ and CD8+ T cell responses to infection with Mycobacterium bovis bacillus Calmette-Guerin (BCG) . After receiving immune, unfractionated T cells or T cell subsets isolated by fluorescence-activated cell sorter, the RAG-1-/- mice were exposed to aerosol BCG, and the bacteria load in the infected organs was examined 4 weeks later . Adoptive immunity was expressed more effectively in the spleens than in the lungs . Although CD4+ or unfractionated T cells protected both lungs and spleens, CD8+ T cells conferred significant protection only in the spleens and not in the lungs . The results confirm that in addition to CD4+, CD8+ T cells also play a role in the prevention of bacterial dissemination . This transfer model may be useful for dissecting T cell responses to mycobacterial infection. Biochemistry, 2000 May 2, 39(17), 4971 - 81 Lysine-73 is involved in the acylation and deacylation of beta-lactamase; Lietz EJ et al.; Lysine 73 is a conserved active-site residue in the class A beta-lactamases, as well as other members of the serine penicillin-sensitive enzyme family; its role in catalysis remains controversial and uncertain . Mutation of Lys73 to alanine in the beta-lactamase from Bacillus licheniformis resulted in a substantial reduction in both turnover rate (k(cat)) and catalytic efficiency (k(cat)/K(m)), and a very significant shift in pK(1) to higher pH in the bell-shaped pH-rate profiles (k(cat)/K(m)) for several penicillin and cephalosporin substrates . The increase in pK(1) is consistent with the removal of the positive ammonium group of the lysine from the proximity of Glu166, to which the acid limb has been ascribed . The alkaline limb of the k(cat)/K(m) vs profiles is not shifted appreciably, as might have been expected if this limb reflected the ionization of Lys73 in the wild-type enzyme . The k(cat)/K(m) at the pH optimum for the mutant was down about 200-fold for penicillins and around 10(4) for cephalosporins, compared to the wild-type, suggesting significant differences in the mechanisms for catalysis of penicillins compared to cephalosporins . Burst kinetics were observed with several substrates assayed with K73A beta-lactamase, indicating an underlying branched-pathway kinetic scheme, and rate-limiting deacylation . FTIR analysis was used to determine whether acylation or deacylation was rate-limiting . In general, acylation was the rate-limiting step for cephalosporin substrates, whereas deacylation was rate-limiting for penicillin substrates . The results indicate that Lys73 plays an important role in both the acylation and deacylation steps of the catalytic mechanism . The effects of this mutation (K73A) indicate that Lys73 does not function as a general base in the catalytic mechanism of beta-lactamase . The existence of bell-shaped pH-rate profiles for the K73A variant suggests that Lys73 is not directly responsible for either limb in such plots . It is likely that both Glu166 and Lys73 are important to each other in terms of maintaining the optimum electrostatic environment for fully efficient catalytic activity to occur. Acta Crystallogr D Biol Crystallogr, 2000 Jun, 56 ( Pt 6), 749 - 50 Crystallization and preliminary X-ray analysis of high-alkaline pectate lyase; Akita M et al.; Pel-15, a high-alkaline pectate lyase (pectate transeliminase; E.C . 4.2.2.2) from Bacillus sp . strain KSM-P15, has been crystallized using the hanging-drop vapour-diffusion method at 277 K . Two different crystal forms were obtained and preliminary X-ray diffraction data were collected from each crystal form at 100 K . Both forms belong to the orthorhombic space group P2(1)2(1)2(1) and contain one molecule per asymmetric unit . The unit-cell parameters of form I are a = 43.2 (2), b = 60.2 (2), c = 82.2 (2) A and those of form II are a = 42.9 (1), b = 43.4 (1), c = 105.9 (3) A . Diffraction data to a resolution of 1.5 A were collected from form II crystals using a synchrotron-radiation source. Microbes Infect, 2000 Apr, 2(4), 431 - 40 Historical perspectives on the etiology of tuberculosis; Barnes DS; Robert Koch's 1882 demonstration that the tubercle bacillus was the true cause of tuberculosis established a new understanding of causation in medicine . This scientific breakthrough set in motion an etiological revolution with vast implications for the control of infectious disease, and its ramifications are still being felt today. Int J Epidemiol, 2000 Apr, 29(2), 369 - 75 Underestimation of Mycobacterium tuberculosis infection in HIV-infected subjects using reactivity to tuberculin and anergy panel; Garcia-Garcia ML et al.; BACKGROUND: This study aimed to evaluate purified protein derivative (PPD) reactivity and its interrelationship with anergy panel and CD4+ lymphocytes in HIV-infected subjects as compared to PPD reactivity in HIV-uninfected individuals in a tuberculosis endemic and high Bacillus Calmette-Guerin (BCG) coverage environment . METHODS: Clients of four Mexico City HIV detection centres were screened for HIV-1 antibodies (ELISA or haemagglutination, Western Blot); reactivity to PPD (Mantoux PPD, 5TU RT-23), Candida (1:1000, 0.1 ml), and tetanus toxoid (10Lf, 0.1 ml); and CD4+ T cells . Active tuberculosis was excluded . Informed consent was obtained . RESULTS: From 5130 clients 1168 subjects were enrolled; of these 801 (68.6%) were HIV positive . Reactivity to PPD among HIV-positive subjects was found in 174 (22%), 261 (32.6%), and 296 (37%), at PPD cutoff levels of > or =10 mm, > or =5 mm, and > or =2 mm as compared to 224 (61%) of 367 HIV-negative individuals' reactors to PPD (> or =10 mm) (P < 0.001) . After exclusion of anergic individuals using two cutoff levels for cutaneous allergens (< or =2 mm and < or =5 mm), PPD reactivity between HIV-infected and uninfected individuals continued to be significantly different . Only HIV-infected individuals with CD4+ T cells > or =500 cells/mm3 had similar reactivity to PPD as HIV-uninfected individuals . Variables associated with PPD reactivity were CD4+ T cell counts, BCG scar, HIV infection and age . CONCLUSIONS: PPD reactivity was useful to diagnose tuberculosis infection only among HIV-infected individuals with CD4+ counts > or =500 cells/mm3 . Among individuals with lower counts, lowering cutoff levels or using anergy panel did not permit comparable reactivity as that observed among HIV-uninfected individuals. Infect Immun, 2000 Jun, 68(6), 3314 - 21 Antigenic equivalence of human T-cell responses to Mycobacterium tuberculosis-specific RD1-encoded protein antigens ESAT-6 and culture filtrate protein 10 and to mixtures of synthetic peptides; Arend SM et al.; The early secreted antigenic target 6-kDa protein (ESAT-6) and culture filtrate protein 10 (CFP-10) are promising antigens for reliable immunodiagnosis of tuberculosis . Both antigens are encoded by RD1, a genomic region present in all strains of Mycobacterium tuberculosis and M . bovis but lacking in all M . bovis bacillus Calmette-Guerin vaccine strains . Production and purification of recombinant antigens are laborious and costly, precluding rapid and large-scale testing . Aiming to develop alternative diagnostic reagents, we have investigated whether recombinant ESAT-6 (rESAT-6) and recombinant CFP-10 (rCFP-10) can be replaced with corresponding mixtures of overlapping peptides spanning the complete amino acid sequence of each antigen . Proliferation of M . tuberculosis-specific human T-cell lines in response to rESAT-6 and rCFP-10 and that in response to the corresponding peptide mixtures were almost completely correlated (r = 0.96, P < 0.0001 for ESAT-6; r = 0.98, P < 0.0001 for CFP-10) . More importantly, the same was found when gamma interferon production by peripheral blood mononuclear cells in response to these stimuli was analyzed (r = 0.89, P < 0.0001 for ESAT-6; r = 0.89, P < 0.0001 for CFP-10) . Whole protein antigens and the peptide mixtures resulted in identical sensitivity and specificity for detection of infection with M . tuberculosis . The peptides in each mixture contributing to the overall response varied between individuals with different HLA-DR types . Interestingly, responses to CFP-10 were significantly higher in the presence of HLA-DR15, which is the major subtype of DR2 . These results show that mixtures of synthetic overlapping peptides have potency equivalent to that of whole ESAT-6 and CFP-10 for sensitive and specific detection of infection with M . tuberculosis, and peptides have the advantage of faster production at lower cost. Biotechnol Appl Biochem, 2000 Jun, 31 ( Pt 3), 171 - 8 Hydrolytic action of alpha-amylase on high-amylose starch of low molecular mass; Rendleman JA Jr; High-amylose starches of low average degree of polymerization (d-p 61-71), formed as fine granules by interaction of Bacillus macerans cyclodextrin glucanotransferase with alpha-cyclodextrin (CD) at 2-70 degrees C, are highly insoluble in water and not gelatinizable under normal cooking conditions (100 degrees C).Samples of CD-derived starches, both cooked and uncooked, were subjected to hydrolysis in vitro by human salivary alpha-amylase at 37 degrees C under conditions chosen to resemble those in the human intestinal lumen . Released low-molecular-mass saccharides were determined quantitatively by HPLC and the results compared with those from similar studies with natural starches . Among uncooked starches, CD-derived starch showed very low reactivity towards alpha-amylase, along with potato starch and a high-amylose hybrid corn starch(64% amylose) . Cooking greatly enhanced reactivity of natural starches, but only moderately increased reactivity of CD-derived starches . Susceptibility to hydrolysis of cooked starches increased in the following general order: CD-derived starch (approximately 100% amylose)<100%corn amylose (isolated by the butan-1-ol method)<hybrid high-amylose corn starch (64-66% amylose)<waxy maize starch (99-100%amylopectin) approximately ordinary corn starch ( approximately 25% amylose)<potato starch (approximately 25% amylose). Cancer Treat Rev, 2000 Jun, 26(3), 169 - 90 Immunomodulation therapy in colorectal carcinoma; Yip D et al.; There has been much progress in the understanding of the relationship between the immune system and colorectal cancer . This has led to the use of immunomodulatory therapy in the adjuvant and palliative treatment of the condition . Although attempts at the use of non-specific immunomodulation with agents such as levamisole, cimetidine, alpha interferon and Bacillus Calmette-Guerin (BCG) have not produced significant clinical benefits when tested in randomized trials in both the adjuvant setting and for metastatic disease, promising results are being obtained with more specific therapy . Edrecolomab {corrected}, a murine monoclonal antibody targeting the 17-1A antigen on malignant colorectal cells has produced a reduction in relapse and mortality rates when used as adjuvant treatment following surgery for Dukes' C colon cancer . Active specific therapy with autologous tumour vaccine administered with BCG has produced similar benefits in Dukes' B cancer . Both 3H1 anti-idiotypic antibody against carcinoembryonic antigen and 105AD7 antibody to gp72 glycoprotein have demonstrated in-vitro and in-vivo immune activation against tumour . Non-randomized studies postulate prolongation of survival using these antibodies in advanced disease . These agents are all currently being tested in randomized studies powered to detect meaningful survival differences and clinical benefit . Immune therapy offers the potential of low toxicity therapy in colorectal cancer and may have a role as an adjunct to conventional chemotherapy . J Org Chem, 2000 Feb 11, 65(3), 878 - 82 Biocatalytic asymmetric hydroxylation of hydrocarbons with the topsoil-microorganism Bacillus megaterium; Adam W et al.; A Bacillus megaterium strain was isolated from topsoil by a selective screening procedure with allylbenzene as a xenobiotic substrate . This strain performed the hydroxylation chemoselectively (no arene oxidation and overoxidized products) and enantioselectively (up to 99% ee) in the benzylic and nonbenzylic positions of a variety of unfunctionalized arylalkanes . Salycilate and phenobarbital, which are potent inducers of cytochrome P-450 activity, changed the regioselectivity of the microbial CH insertion, without an effect on the enantioselectivity . The biotransformation conditions were optimized in regard to product yield and enantioselectivity by variation of the oxygen-gas supply and the time of the substrate addition . The different product distributions (alpha- versus beta-hydroxylated product) that are obtained on induction of cytochrome P-450 enzyme activity demonstrate the involvement of two or more hydroxylating enzymes with distinct regioselectivities in this biotransformation . An oxygen-rebound mechanism is assumed for the cytochrome P-450-type monooxygenase activity, in which steric interactions between the substrate and the enzyme determine the preferred face of the hydroxy-group transfer to the radical intermediate. FEBS Lett, 2000 May 12, 473(2), 227 - 32 Enhanced toxicity of Bacillus thuringiensis Cry3A delta-endotoxin in coleopterans by mutagenesis in a receptor binding loop; Wu SJ et al.; We used site-directed mutagenesis to modify the Bacillus thuringiensis cry3A gene in amino acid residues 350-354 . Two mutant toxins, A1 (R(345)A,Y(350)F,Y(351)F) and A2 (R(345)A,DeltaY(350), DeltaY(351)), showed significantly improved toxicity against Tenebrio molitor (yellow mealworm) . The mutant toxin A1 was also more potent against both Leptinotarsa decemlineata (Colorado potato beetle) and Chrysomela scripta (cottonwood leaf beetle), while A2 displayed enhanced toxicity only in L . decemlineata . Competitive binding assays of L . decemlineata brush border membrane vesicles (BBMV) revealed that binding affinities for the A1 and A2 mutant toxins were ca . 2.5-fold higher than for the wild-type Cry3 toxin . Similar binding assays with C . scripta BBMV revealed a ca . 5-fold lower dissociation rate for the A1 mutant as compared to that of Cry3A. J Membr Biol, 2000 May 15, 175(2), 115 - 22 Kinetic properties of the channels formed by the bacillus thuringiensis insecticidal crystal protein Cry1C in the plasma membrane of Sf9 cells; Guihard G et al.; Spectrofluorimetric measurements were conducted to quantify, in real-time, membrane permeability changes resulting from the treatment of Sf9 insect cells (Spodoptera frugiperda, Lepidoptera) with different Bacillus thuringiensis Cry insecticidal proteins . Coumarin-derived CD222 and Merocyanin-540 probes were respectively used to monitor extracellular K(+) and membrane potential variations upon Sf9 cells incubation with Cry toxins . Our results establish that Cry1C induces, after a delay, the depolarization of the cell membrane and the full depletion of intracellular K(+) . These changes were not observed upon Sf9 cells treated with Cry1A family toxins . Both the rate of the K(+) efflux and the delay before its onset were dependent on toxin concentration . Both parameters were sensitive to temperature but only the delay was affected by pH . Cry1C-induced K(+) efflux was inhibited by lanthanum ions in a dose-dependent manner . This study provides the first kinetic and quantitative characterization of the ion fluxes through the channels formed by a Cry toxin in the plasma membrane of a susceptible insect cell line. J Biol Chem, 2000 Aug 4, 275(31), 23602 - 7 Insertion and organization within membranes of the delta-endotoxin pore-forming domain, helix 4-loop-helix 5, and inhibition of its activity by a mutant helix 4 peptide; Gerber D et al.; The pore-forming domain of Bacillus thuringiensis Cry1Ac insecticidal protein comprises of a seven alpha-helix bundle (alpha1-alpha7) . According to the "umbrella model," alpha4 and alpha5 helices form a hairpin structure thought to be inserted into the membrane upon binding . Here, we have synthesized and characterized the hairpin domain, alpha4-loop-alpha5, its alpha4 and alpha5 helices, as well as mutant alpha4 peptides based on mutations that increased or decreased toxin toxicity . Membrane permeation studies revealed that the alpha4-loop-alpha5 hairpin is extremely active compared with the isolated helices or their mixtures, indicating the complementary role of the two helices and the need for the loop for efficient insertion into membranes . Together with spectrofluorometric studies, we provide direct evidence for the role of alpha4-loop-alpha5 as the membrane-inserted pore-forming hairpin in which alpha4 and alpha5 line the lumen of the channel and alpha5 also participates in the oligomerization of the toxin . Strikingly, the addition of the active alpha4 mutant peptide completely inhibits alpha4-loop-alpha5 pore formation, thus providing, to our knowledge, the first example that a mutated helix within a pore can function as an "immunity protein" by directly interacting with the segments that form the pore . This presents a potential means of interfering with the assembly and function of other membrane proteins as well. Rev Inst Med Trop Sao Paulo, 2000 Mar-Apr, 42(2), 111 - 3 Chromobacterium violaceum infection in Brazil . A case report; Martinez R et al.; We report the second case of infection with Chromobacterium violaceum that occurred in Brazil . A farm worker living in the State of Sao Paulo presented fever and severe abdominal pain for four days . At hospitalization the patient was in a toxemic state and had a distended and painful abdomen . Chest X-ray and abdominal ultrasound revealed bilateral pneumonia and hypoechoic areas in the liver . The patient developed failure of multiple organs and died a few hours later . Blood culture led to isolation of C . violaceum resistant to ampicillin and cephalosporins and sensitive to chloramphenicol, tetracyclin, aminoglicosydes, and ciprofloxacin . Autopsy revealed pulmonary microabscesses and multiple abscesses in the liver . The major features of this case are generally observed in infections by C . violaceum: rapid clinical course, multiple visceral abscesses, and high mortality . Because of the antimicrobial resistance profile of this Gram-negative bacillus, for appropriate empirical antibiotic therapy it is important to consider chromobacteriosis in the differential diagnosis of severe community infections in Brazil. J Bacteriol, 2000 Jun, 182(11), 3045 - 54 Expression and characterization of the chitin-binding domain of chitinase A1 from Bacillus circulans WL-12; Hashimoto M et al.; Chitinase A1 from Bacillus circulans WL-12 comprises an N-terminal catalytic domain, two fibronectin type III-like domains, and a C-terminal chitin-binding domain (ChBD) . In order to study the biochemical properties and structure of the ChBD, ChBD(ChiA1) was produced in Escherichia coli using a pET expression system and purified by chitin affinity column chromatography . Purified ChBD(ChiA1) specifically bound to various forms of insoluble chitin but not to other polysaccharides, including chitosan, cellulose, and starch . Interaction of soluble chitinous substrates with ChBD(ChiA1) was not detected by means of nuclear magnetic resonance and isothermal titration calorimetry . In addition, the presence of soluble substrates did not interfere with the binding of ChBD(ChiA1) to regenerated chitin . These observations suggest that ChBD(ChiA1) recognizes a structure which is present in insoluble or crystalline chitin but not in chito-oligosaccharides or in soluble derivatives of chitin . ChBD(ChiA1) exhibited binding activity over a wide range of pHs, and the binding activity was enhanced at pHs near its pI and by the presence of NaCl, suggesting that the binding of ChBD(ChiA1) is mediated mainly by hydrophobic interactions . Hydrolysis of beta-chitin microcrystals by intact chitinase A1 and by a deletion derivative lacking the ChBD suggested that the ChBD is not absolutely required for hydrolysis of beta-chitin microcrystals but greatly enhances the efficiency of degradation. Mikrobiologiia, 2000 Jan-Feb, 69(1), 29 - 33 {Effect of culture media components on accumulation of glutaminyl endopeptidase in culture liquid of Bacillus intermedius 3-19}; Shakirov EV et al.; The effect of nutrients and growth conditions on the accumulation of glutamyl endopeptidase in the culture liquid of Bacillus intermedius 3-19 was studied . Glucose and other readily metabolizable carbon sources were found to suppress the production of the enzyme, while inorganic phosphate and ammonium cations enhanced it . Protein substrates, such as casein, gelatin, and hemoglobin, did not affect enzyme production . Some bivalent cations (Ca2+, Mg2+, Co2+) increased the production of glutamyl endopeptidase, but others (Zn2+, Fe2+, Cu2+) acted in the opposite way . The rate of enzyme accumulation in the culture liquid increased as the growth rate of the bacterium decreased, so that the maximum enzyme activity was observed in the stationary growth phase . Based on the results of this investigation, an optimal medium for the maximum production of glutamyl endopeptidase by B . intermedius 3-19 was elaborated. J Struct Biol, 2000 May, 130(1), 1 - 9 Structure of apo-glyceraldehyde-3-phosphate dehydrogenase from Palinurus versicolor; Shen YQ et al.; d-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) shows cooperative properties for binding coenzymes . The structure of apo-GAPDH from Palinurus versicolor has been solved at 2.0 A resolution by X-ray crystallography . The final model gives a crystallographic R factor of 0.178 in the resolution range 8 to 2 A . The structural comparison with holo-GAPDH from the same species reveals a conformational change induced by coenzyme binding similar to that observed in Bacillus stearothermophilus GAPDH but to a lesser extent . The differences in magnitude during the apo-holo transition between these two enzymes were analyzed with respect to the change of the amino acid composition in the coenzyme binding pocket . In the crystalline state of apo-GAPDH, the overall structures of the subunits are similar to each other; however, significant differences in temperature factors and minor differences in domain rotation upon coenzyme binding were observed for different subunits . These structural features are discussed in relation to the environmental asymmetry of crystallographically independent subunits . Anal Biochem, 2000 Apr 10, 280(1), 159 - 65 Two continuous spectrophotometric assays for methionine aminopeptidase; Zhou Y et al.; Two spectrophotometric assays have been developed for methionine aminopeptidases (MetAPs) . The first method employs a thioester substrate which, upon enzymatic removal of the N-terminal methionine, generates a free thiol group . The released thiol is quantitated using Ellman's reagent . The MetAP reaction is conveniently monitored on a UV-VIS spectrophotometer in a continuous fashion, with the addition of an excess of Ellman's reagent into the assay reaction . Two tripeptide analogues were synthesized and found to be excellent substrates of both Escherichia coli MetAP and human MetAP2 (k(cat)/K(M) = 2.8 x 10(5) M(-1) s(-1) for the most reactive substrate) . In the second assay method, the MetAP reaction is coupled to a prolyl aminopeptidase reaction using Met-Pro-p-nitroanilide as substrate . MetAP-catalyzed cleavage of the N-terminal methionine produces prolyl-p-nitroanilide, which is rapidly hydrolyzed by the prolyl aminopeptidase from Bacillus coagulans to release a chromogenic product, p-nitroaniline . This allows the MetAP reaction to be continuously monitored at 405 nm on a UV-VIS spectrophotometer . The assays have been applied to determine the pH optima and kinetic constants for the E . coli and human MetAPs as well as to screen MetAP inhibitors . These results demonstrate that the current assays are convenient, rapid, and sensitive methods for kinetic studies of MetAPs and effective tools for screening MetAP inhibitors. Biosci Biotechnol Biochem, 2000 Mar, 64(3), 670 - 3 An automated system for genome analysis to support microbial whole-genome shotgun sequencing; Sakiyama T et al.; We developed a semi-automated genome analysis system called GAMBLER in order to support the current whole-genome sequencing project focusing on alkaliphilic Bacillus halodurans C-125 . GAMBLER was designed to reduce the human intervention required and to reduce the complications in annotating thousands of ORFs in the microbial genome . GAMBLER automates three major routines: analyzing assembly results provided by genome assembler software, assigning ORFs, and homology searching . GAMBLER is equipped with an interface for convenience of annotation . All processes and options are manipulatable through a WWW browser that enables scientists to share their genome analysis results without choosing computer platforms. Biosci Biotechnol Biochem, 2000 Mar, 64(3), 665 - 9 Efficient production of N-terminally truncated biologically active human interleukin-6 by Bacillus brevis; Shiga Y et al.; cDNAs encoding human interleukin 6 (hIL-6) and its variants lacking the N-terminal Pro and Pro-Val-Pro-Pro, respectively, were expressed in Bacillus brevis by using the signal peptide fusion approach . The presence of Pro at the N-terminus of the mature protein hindered the action of the Bacillus brevis signal peptidase . hIL-6 lacking the N-terminal Pro-Val-Pro-Pro was most efficiently secreted in a biologically active form and accumulated in the culture medium to a level of 200 mg per liter, which is the highest level reported for the bacterial secretion of hIL-6. Appl Microbiol Biotechnol, 2000 Apr, 53(4), 390 - 5 Soybean-milk-coagulating activity of Bacillus pumilus derives from a serine proteinase; Aoyama M et al.; A proteolytic enzyme from Bacillus pumilus strain TYO-67, which was able to coagulate the protein in soybean milk, was characterized enzymologically . The optimum pH and temperature for its activities were 9.0 and 50 degrees C, respectively . The enzyme was strongly believed to be a serine proteinase because it was completely inhibited by the addition of diisopropyl fluorophosphate or phenylmethanesulfonyl fluoride . Hammerstein milk casein, cytochrome c and soybean protein were good substrates for the enzyme . Seven cleavages were detected using the oxidized insulin B-chain as peptide substrate for the proteolytic specificity test of the serine proteinase from B . pumilus . The bonds most susceptible to the action of the serine proteinase from B . pumilus were Leu-15-Tyr-16 . The mode of action on soybean milk protein by the enzyme from B . pumilus was also investigated . The acidic subunit in glycinin and the alpha-, alpha- and beta-subunits in beta-conglycinin were degraded during the enzyme reaction . However, the basic subunit in glycinin could not be degraded by the enzyme . The formation of coagula in soybean milk caused by the serine proteinase from B . pumilus was mainly due to the hydrophobic interaction. Med Dosw Mikrobiol, 1999, 51(3-4), 347 - 55 {Bacterial infections of skin and soft tissues in filariasis}; Swoboda-Kopec E et al.; Adenolymphangitis is a common occurrence in filarial lymphedema . Damage to the lymphatics and lymph nodes by F . bancrofti is followed by obliteration of lymph vessels and lymph stasis . Obstruction of lymphatics prevents the bacteria penetrating skin to be evacuated with lymph stream to regional lymph nodes . Colonization of dermis, subcutis and lymphatics evokes clinical symptoms of adenolymphangitis . The question arises which strains of bacteria are responsible for the acute and chronic types of adenolymphangitis . The most probable strains responsible for this condition belong to the cocci and probably the bacillus strains. Am J Surg, 2000 Feb, 179(2A Suppl), 45S - 50S Role of aztreonam in the treatment of nosocomial pneumonia in the critically ill surgical patient; Boucher BA; In 1995 the American Thoracic Society issued an official consensus statement on the treatment of hospital-acquired pneumonia (HAP) . Classes of antimicrobials included in the list of antimicrobials deemed to be suitable for the empiric treatment of severe HAP were the aminoglycosides, quinolones, antipseudomonal penicillins, carbapenems, and beta-lactam/beta-lactamase inhibitor combinations . Aztreonam, a monobactam, was also listed and is unique among these agents based on its spectrum of activity being limited to the gram-negative bacillary bacteria combined with an excellent safety profile . This review focuses on the role of aztreonam in the treatment of nosocomial pneumonia in the critically ill patient.A review of the literature was performed using PubMed and secondary literature sources as to the clinical efficacy of aztreonam in the treatment of lower respiratory tract infections as well as its pharmacokinetic and safety profiles . An analysis of aztreonam's potential pharmacoeconomic advantages compared with other agents was also performed.Numerous studies have documented that aztreonam has effectiveness that is equal or superior to that of other suitable antibiotics in the treatment of nosocomial pneumonia . Its excellent safety profile makes it a particularly attractive agent compared with the aminoglycosides . Considering the potential costs of bacterial resistance from the use of broader-spectrum alternatives, a case can be made that aztreonam is a pharmacoeconomically sound choice as well. Biochemistry, 2000 May 16, 39(19), 5775 - 87 Mutational analysis of the epimerization domain in the initiation module PheATE of gramicidin S synthetase; Stachelhaus T et al.; The epimerase (E) domain of the three-domain (ATE) initiation module of Bacillus brevis gramicidin S synthetase equilibrates the Calpha configuration of the phenylalanyl moiety presented as Phe-S-4'-phosphopantetheine-modified (Ppant) acyl enzyme . Mutants at 22 residues of this E domain that are conserved across the approximately 450 residue E domains of nonribosomal peptide synthetases were constructed, and the PheATE derivatives expressed in Escherichia coli as C-terminal His tag fusions and then purified and assayed for three activities: (1) the L-Phe Calpha-{(3)H} exchange to solvent, (2) the rate of approach to D-Phe/L-Phe-S-Ppant acyl enzyme equilibrium from either L- or D-Phe, and (3) the rate of Phe-Pro dipeptidyl-S-Ppant enzyme formation with the downstream ProCAT module . We found that for wild-type PheATE epimerization is much faster than subsequent condensation, leading to a 1.9:1 ratio of D-Phe-S-Ppant/L-Phe-S-Ppant acyl enzyme . Only D-Phe is then transferred to yield D-Phe-L-Pro-S-Ppant ProCAT acyl enzyme . Among the mutants generated, three PheATE constructs, H753A, D757S, and Y976A, showed no detectable Calpha-(3)H washout, while E892A and R896A were among a larger set partially impaired . All these mutants were dramatically impaired in approach to D-Phe/L-Phe-S-Ppant equilibrium from either D- or L-Phe, while another construct, D767S, was asymmetrically impaired only for D-to-L-Phe direction . In the D-Phe-L-Pro dipeptidyl-S-Ppant condensation assay, the H753A and E892A forms of PheATE were only slightly active from L-Phe but unimpaired from D-Phe; N975A epimerizes faster than Y976A from L-Phe . When the chirality of the Phe-Pro-diketopiperazine released product was analyzed the D,L/L,L ratio from wild-type PheATE and ProCAT was 98:2 . From E892A and N975A it was comparably 95:5 and 92:8, but H753A and Y976A yielded 56% of the L,L-product, reflecting a gain of function to transfer L-Phe . The 98:2 preference of wild-type PheATE for D-Phe transfer reflects the kinetically controlled stereopreference of the condensation (C) domain of ProCAT for the D-Phe-S-Ppant donor substrate . It may be that other NRPS C domains immediately downstream of E domains will likewise be D-selective. J Biol Chem, 2000 May 12, 275(19), 14423 - 31 Bivalent sequential binding model of a Bacillus thuringiensis toxin to gypsy moth aminopeptidase N receptor; Jenkins JL et al.; Specificity for target insects of Bacillus thuringiensis insecticidal Cry toxins is largely determined by toxin affinity for insect midgut receptors . The mode of binding for one such toxin-receptor complex was investigated by extensive toxin mutagenesis, followed by real-time receptor binding analysis using an optical biosensor (BIAcore) . Wild-type Cry1Ac, a three-domain, lepidopteran-specific toxin, bound purified gypsy moth (Lymantria dispar) aminopeptidase N (APN) biphasically . Site 1 displayed fast association and dissociation kinetics, while site 2 possessed slower kinetics, yet tighter affinity . We empirically determined that two Cry1Ac surface regions are involved in in vivo toxicity and APN binding . Mutations within domain III affected binding rates to APN site 1, whereas mutations in domain II affected binding rates to APN site 2 . Furthermore, domain III contact is completely inhibited in the presence of N-acetylgalactosamine, indicating loss of domain III binding eliminates all APN binding . Based upon these observations, the following model is proposed . A cavity in lectin-like domain III initiates docking through recognition of an N-acetylgalactosamine moiety on L . dispar APN . Following primary docking, a higher affinity domain II binding mechanism occurs, which is critical for insecticidal activity. J Urol, 2000 Jun, 163(6), 1697 - 701 Recurrence and progression of stage T1, grade 3 transitional cell carcinoma of the bladder following intravesical immunotherapy with bacillus Calmette-Guerin; Brake M et al.; PURPOSE: We prospectively examined the incidence of recurrence and progression in patients with stage pT1, grade 3 carcinoma of the bladder following complete transurethral resection of the bladder tumor and adjuvant immunotherapy with bacillus Calmette-Guerin (BCG) . MATERIALS AND METHODS: Between July 1987 and March 1999, 123 patients presenting to our clinic with superficial urothelial carcinoma (stage pT1, grades 1 to 3) received adjuvant intravesical immunotherapy with BCG after histologically confirmed complete transurethral tumor resection . Disease was stage pT1, grade 3 in 44 patients (36%) . Median followup was 28 months (mean 43, range 5 to 141) . RESULTS: Of the patients 36 (82%) with bladder preservation remained tumor-free during followup after 1 or 2 cycles of BCG . Superficial tumor recurred in 5 patients (11%) and muscle invasive progression was noted in 7 (16%) . Radical cystectomy was performed in 4 cases (9%) . Of the patients 5 (11%) died of cancer . Tumor-free survival for all patients was 89% (39 of 44) . CONCLUSIONS: Adjuvant immunotherapy with BCG after complete transurethral resection of bladder tumor represents a highly effective primary treatment of stage pT1, grade 3 carcinoma of the bladder . Immediate radical cystectomy does not appear necessary. Can J Gastroenterol, 2000 Apr, 14(4), 333 - 6 Hepatitis in disseminated bacillus Calmette-Guérin infection; Gottke MU et al.; Local immunotherapy with an attenuated live strain of Mycobacterium bovis, bacillus Calmette-Guerin (BCG), is an effective and frequently used treatment for in situ transitional cell carcinoma (TCC) of the bladder . Success rates are high, and serious side effects are infrequent but can affect every organ system . A 79-year-old patient with recently diagnosed TCC who was treated with intravesical BCG for a recurrence after initial surgical treatment is reported . After unsuccessful attempts at bladder catheterization with the creation of a false passage for his third treatment, BCG was instilled via a suprapubic catheter the same day and again a week later . Two weeks after the third BCG instillation, the patient presented with profound lethargy and weakness to the point of not being able to get up out of a chair . He was febrile, anorexic, icteric and had hepatosplenomegaly . Disseminated BCG infection was suspected on the basis of history, clinical examination and a liver biopsy that showed noncaseating granulomatous hepatitis . Empirical treatment was started with antituberculous combination therapy . A short course of an oral corticosteroid was given . Clinical improvement was marked and sustained so that the patient could be discharged home for the full six-month course of his treatment . Disseminated BCG infection with granulomatous hepatitis can be severe and life-threatening in cases where a large intravascular inoculum of BCG may have been given inadvertently. Dakar Med, 1999, 44(1), 123 - 5 {Hepatic tuberculosis of the pseudotumoral form}; Diouf ML et al.; The authors report a case of a 30 years old immunocompetent woman with liver tuberculosis with an unusual pseudotumoral presentation and secondary occurrence of abcedation with cutaneous fistulization . The diagnosis was based on the bacteriological positivity for acido-alcoolo resistant bacillus in pus of the abscess obtained by ponction guided by ultrasonographic examination . The authors emphasize in the differential diagnosis with the other causes of liver abscess (amibiasis and pyogenic microorganisms) and liver carcinoma . They also note the importance of the function guided by ultrasonographic examination permitting histologic and bacte+eriologic study in the diagnosis of this unusual presentation of liver tuberculosis . The patient was treated by antibacillary antibiotics and evacuation function . The prognostic was good with 6 months of follow-up. Dakar Med, 1998, 43(2), 170 - 3 {Characterization of entomopathogenic Bacillus samples isolated in Senegal and study of their toxicity for malaria vectors}; Aidara-Kane A et al.; A screening program developed in Senegal to isolate new strains of entomopathogenic Bacillus has led to the isolation of 194 strains of Bacillus thuringiensis and 9 strains of Bacillus sphaericus from various sites and insect samples . The characterization of these strains regarding their H serotype, their crystal composition and their toxicity against mosquitoes (Culex pipiens, Aedes aegypti and Anopheles stephensi) has led to the isolation of 27 mosquitocidal strains . As malaria is an important public health problem in Senegal, these strains were more completely characterized looking for their toxicity against the two major malaria vectors in Senegal: Anopheles gambiae and Anopheles arabiensis. Biotechnol Bioeng, 2000 Jun 5, 68(5), 557 - 62 Production of recombinant L-leucine dehydrogenase from Bacillus cereus in pilot scale using the runaway replication system E . coli{pIET98}; Ansorge MB et al.; A method for the production of recombinant L-leucine dehydrogenase from Bacillus cereus in pilot scale is described employing the temperature induced runaway replication vector pIET98 and the Escherichia coli host strain BL21 . Fed-batch cultivation using a semi-synthetic high-cell densitiy medium was adjusted in 5-L scale to yield a constant growth rate of 0,17 h(-1) and a final cell concentration of 27 g dry weight/L by exponentially increasing the nutrient supply . Runaway replication and thus, LeuDH expression was induced during the feeding phase by increasing the cultivation temperature to 41 degrees C yielding a specific enzyme activity of 110 U/mg, which corresponds to 30% of the soluble cell protein . The cultivation was terminated when the dissolved oxygen content fell below 10% saturation . The final volume activity was 600,000 U/L cultivation . No change in growth, cell density, or expression activity was observed scaling up the cultivation volume to 200 L . Thus, 120,000,000 units L-leucine dehydrogenase were obtained from one cultivation . The purification of L-leucine dehydrogenase to homogeneity was carried out by heat denaturation, liquid-liquid extraction, gel filtration, and anion-exchange chromatography to give pure enzyme in 65% yield . The integrity of the recombinant enzyme was tested measuring the molecular weight and determining the N-terminal amino acid sequence . Surg Today, 2000, 30(4), 383 - 5 Splenic vein occlusion secondary to tuberculous lymphadenitis at the splenic hilum: report of a case; Takeuchi H et al.; We report a patient with splenic vein occlusion (SVO) secondary to tuberculosis . A 17-year-old male patient with mild epigastric pain and splenomegaly was found to have gastric varices by gastroscopy, and SVO by selective angiography . At operation, the splenic vein was occluded by hard fibrous tissue at the splenic hilum, and thus a splenectomy was performed . A microscopic examination of the tissue revealed caseous necrosis surrounded by epithelioid cells and Langhans-type giant cells . Although there were no other findings suggesting intestinal tuberculosis, it seemed that tuberculous lymphadenitis of the splenic hilum most likely caused the occlusion of the splenic vein . Because specific tests for tuberculosis were negative in both immunohistochemical staining for bacille Calmette-Guerin and polymerase chain reaction of DNA for Mycobacterium tuberculosis, the time of infection was assumed to have occurred a long time before . SVO can sometimes be seen in pancreatic diseases, but this patient with tuberculosis appears to be the first such reported case in the English literature. Carbohydr Res, 2000 Apr 7, 325(2), 120 - 31 Regioselective synthesis of p-nitrophenyl glycosides of beta-D-galactopyranosyl-disaccharides by transglycosylation with beta-D-galactosidases; Zeng X et al.; The beta-D-galactosidase from porcine liver induced regiospecific transglycosylation of beta-D-galactose from beta-D-Gal-OC6H4NO2-o to OH-6 of, respectively, p-nitrophenyl glycoside acceptors of Gal, GlcNAc and GalNAc to afford beta-Gal-(1-->6)-alpha-Gal-OC6H4NO2-p, beta-Gal-(1--> 6)-beta-Gal-OC6H4NO2-p, beta-Gal-(1-->6)-alpha-GalNAc-OC6H4NO2-p, beta-Gal-(1-->6)-beta-GalNAc-OC6H4NO2-p, beta-Gal-(1-->6)-alpha-GlcNAc-OC6H4NO2-p, and beta-Gal-(1-->6)-beta-GlcNAc-OC6H4NO2-p . The enzyme showed much higher transglycosylation activity for the alpha-glycoside acceptors than the corresponding beta-glycoside acceptors . The regioselectivity of the beta-D-galactosidase from Bacillus circulans ATCC 31382 greatly depended on the nature of the acceptor . When alpha-D-GalNAc-OC6H4NO2-p and alpha-D-GlcNAc-OC6H4NO2-p were used as acceptors, the enzyme showed high potency for regioselective synthesis of beta-Gal-(1-->3)-alpha-GalNAc-OC6H4NO2-p and beta-Gal-(1-->3)-alpha-GlcNAc-OC6H4NO2-p in high respective yields of 75.9 and 79.3% based on the acceptors added . However, replacement of beta-D-Gal-OC6H4NO2-p by beta-D-GalNAc-OC6H4NO2-p did change the direction of galactosylation . The enzyme formed regioselectively beta-Gal-(1-->6)-beta-Gal-OC6H4NO2-p with (beta-Gal-1-->(6-beta-Gal-1-->)n6-beta-Gal-OC6H4NO2-p, n = 1-4) . No beta-(1-->3)-linked product was detected during the reaction . Use of the two readily available beta-D-galactosidases facilitates the preparation of (1-->3)- and (1-->6)-linked disaccharide glycosides of beta-D-Gal-GalNAc and beta-D-Gal-GlcNAc. Scand J Immunol, 2000 May, 51(5), 485 - 90 Modulation of neutrophil influx with cell adhesion molecule specific antibodies during nonspecific and immune mediated inflammatory reactions; Goncalves AS et al.