J Biol Chem, 1992 Nov 5, 267(31), 22108 - 14 Catalytic properties of the cloned amylase from Bacillus licheniformis; Kim IC et al.; A gene encoding a new amylolytic enzyme of Bacillus licheniformis (BLMA) has been cloned, and we characterized the enzyme expressed in Escherichia coli . The genomic DNA of B . licheniformis was double-digested with EcoRI and BamHI and ligated the pBR322 . The transformed E . coli was selected by its amylolytic activity, which carries the recombinant plasmid pIJ322 containing a 3.5-kilobase fragment of B . licheniformis DNA . The purified enzyme encoded by pIJ322 was capable of hydrolyzing pullulan and cyclodextrin as well as starch . It was active over a pH range of 6-8 and its optimum temperature was 50 degrees C . The molecular weight of the enzyme was 64,000, and the isoelectric point was 5.4 . It degraded soluble starch by cleaving maltose units preferentially but did not attack alpha-1,6-linkage . The enzyme also hydrolyzed pullulan to panose units exclusively . In the presence of glucose, however, it transferred the panosyl moiety to glucose with the formation of alpha-1,6-linkage . The specificity of transferring activity is evident from the result of the maltosyl-transferring reaction which produces isopanose from maltotriose and glucose . The molecular structure of the enzyme deduced from the nucleotide sequence of the clone maintains limited similarity in the conserved regions to the other amylolytic enzymes. Microbiologia, 1992 Nov, 8(2), 115 - 8 {Detection of Bacillus larvae in mixed populations of bacterial spores from larval remains}; Alippi AM; An accurate laboratory technique for the detection of Bacillus larvae from larval remains of Apis mellifera with mixed bacterial spore populations was developed . The incorporation of nalidixic acid to the culture medium (3 micrograms/ml) was a satisfactory procedure for the separation of Bacillus larvae strains from Bacillus alvei motile colonies. Mol Biol (Mosk), 1992 Nov-Dec, 26(6), 1338 - 49 {Extracellular alkaline ribonuclease of Bacillus thuringiensis var . subtoxicus}; Dement'ev AA et al.; Intraspecific selection of Bacillus thuringiensis strains producing extracellular alkaline ribonucleases was carried out . Subtoxicus subspecies with increased expression of the enzyme was detected . A method was developed to isolate preparative amounts of homogeneous extracellular RNase of B . thuringiensis var . subtoxicus . The physico-chemical and catalytic properties of the enzyme was studied and compared with extracellular RNases of others Bacillus species . The conclusion about the structural and evolutional conservation of Bacillus extracellular RNases was drawn. Immunobiology, 1992 Nov, 186(3-4), 230 - 40 Induction of an increased number of dendritic cells in the peritoneal cavity of rats by intraperitoneal administration of Bacillus Calmette-Guérin; van Vugt E et al.; Recently we described the presence of a small number of DC among the peritoneal cells of steady state rats . These DC had the same morphological characteristics and a similar antigen-presenting capacity as DC isolated from the spleen . This study shows that in the peritoneal cavity, which is a non-lymphoid microenvironment, the number of DC increases after i.p . administration of BCG . Next to this relatively small influx of DC, the approximately three-fold increase of the total number of cells is predominantly caused by an enormous influx of neutrophilic granulocytes, and to a lesser extent by an influx of macrophages . The phenotype and the antigen-presenting capacity of peritoneal DC has not changed, while the number of Ia-positive M phi has increased . Nevertheless, due to a suppressive effect of the peritoneal M phi, the total peritoneal cell suspension is no longer capable of presenting antigen. Appl Environ Microbiol, 1992 Nov, 58(11), 3779 - 83 Alteration of specific activity and stability of thermostable neutral protease by site-directed mutagenesis; Kubo M et al.; On the basis of three-dimensional information, many amino acid substitutions were introduced in the thermostable neutral protease (NprM) of Bacillus stearothermophilus MK232 by site-directed mutagenesis . When Glu at position 143 (Glu-143), which is one of the proposed active sites, was substituted for by Gln and Asp, the proteolytic activity disappeared . F114A (Phe-114 to Ala), Y110W (Tyr-110 to Trp), and Y211W (Tyr-211 to Trp) mutant enzymes had higher activity (1.3- to 1.6-fold) than the wild-type enzyme . When an autolysis site, Tyr-93, was replaced by Gly and Ser, the remaining activities of those mutant enzymes were higher than that of the wild-type enzyme. Clin Infect Dis, 1992 Nov, 15 Suppl 1, S268 - 73 Evaluation of new anti-infective drugs for the treatment of gastritis and peptic ulcer disease associated with infection by Helicobacter pylori . Infectious Diseases Society of America and the Food and Drug Administration; Murray DM et al.; Helicobacter pylori is a gram-negative, microaerophilic, spiral bacillus . Infection by this organism is currently believed to be the major cause of type B gastritis . Inflammation and infection may persist for years in the absence of therapeutic intervention . There is currently no approved antimicrobial therapy for gastritis . Clinical investigations have shown that combination regimens including bismuth salts and antimicrobial drugs result in the relief of symptoms, the resolution of histologic evidence of gastritis, the eradication of H . pylori, high rates of ulcer healing, and lower rates of ulcer relapse than have been found with other therapies (antacids and H2 antagonists) . A randomized, double-blind, placebo-controlled study design is recommended for evaluation of new therapies . Study participants should have their progress monitored by endoscopy performed at enrollment, at completion of therapy, and 3 months thereafter . Assessment of microbiological outcome is paramount for final evaluation of the patient. J Parenter Sci Technol, 1992 Nov-Dec, 46(6), 215 - 25 Vaporized hydrogen peroxide sterilization of freeze dryers; Johnson JW et al.; The feasibility of using vapor hydrogen peroxide (VHP) as an alternative to steam sterilization has been examined using a pilot plant freeze dryer equipped with a prototype vapor generator . Specific objectives of the study discussed in this presentation were to: 1 . Identify critical process variables affecting the lethality of VHP to Bacillus stearothermophilus spores, particularly within dead legs in the system . 2 . Measure the efficacy of system degassing after sterilization . 3 . Determine the effect of repeated sterilization cycles on the integrity of elastomeric components of the freeze dryer . Penetration of adequate concentrations of hydrogen peroxide vapor into small diameter piping, such as tubing connected to pressure gauges, is the most challenging aspect of VHP sterilization of freeze dryers . Prior to equipment modifications, spore strips placed within such dead legs remained positive irrespective of the number of gas/degas pulses and system pressure . Equipment modifications necessary to effect complete kill of biological indicators placed in system dead legs is discussed . Results of this study support the conclusion that vaporized hydrogen peroxide shows promise as an alternative sterilization method for freeze dryers. J Infect, 1992 Nov, 25(3), 291 - 7 Bacillus cereus meningitis in two neurosurgical patients: an investigation into the source of the organism; Barrie D et al.; Two patients developed Bacillus cereus meningitis following neurosurgery . During the subsequent investigation into the source of the organism, linen was discovered to be heavily contaminated with B . cereus . No other prolific source of the organism was found . It seems probable that lint from contaminated fabric was the vehicle of transmission of the organism during extended surgery . Linen should be considered as a possible source of B . cereus infection. Mol Reprod Dev, 1992 Nov, 33(3), 235 - 42 Genome exclusion and gametic DAPI-DNA content in the hybridogenetic Bacillus rossius-grandii benazzii complex (Insecta Phasmatodea); Tinti F et al.; Among Sicilian stick insects, two hybridogenetic complexes have been discovered: Bacillus rossius-grandii benazzii and B . rossius-grandii grandii, which also produce androgenetic offspring . The egg maturation of the former is analyzed here through DAPI fluorometry, which, besides the assessment of the meiotic stages, also allows their DNA measurements and the analysis of sperm-head evolution into male pronuclei in these polyspermic eggs . Hybridogenetic eggs undergo an extrasynthesis of chromosomes, because two groups of n autobivalents (4C each) are segregated at metaphase 1st; the two groups must correspond to the pure parental species haplosets . Then the grandii chromosomes degenerate (1st polar body), while the rossius chromosomes divide further to produce two groups of n autodiads (2C each); one of them degenerates (2nd polar body), and the other is ready to perform syngamy (female pronucleus) . Meanwhile, several B . grandii sperm evolve into male pronuclei by doubling their DNA (from 1C to 2C content) and assuming an interphase nucleus appearance . If regular mixis occurs, the F1 hybrid constitution is restored but, if it fails, a fusion between two sperms may occur, originating fully paternal descendants (natural androgenesis) . The genome exclusion mechanism of stick-insect hybridogens appears to be more primitive than those observed in the already known hybridogenetic complexes of Poeciliopsis and Rana esculenta . Unfertilized eggs of hybridogens are capable of self-activation, but the cytology of the related clonally reproducing B . whitei indicates that its parthenogenetic mechanism stems from the hybridization event (hybrid theory) rather than from tychoparthenogenetic potentialities (spontaneous theory). Ultrastruct Pathol, 1992 Nov-Dec, 16(6), 629 - 40 Immunohistochemical and electron microscopic profiles of cutaneous Kaposi's sarcoma and bacillary angiomatosis; Kostianovsky M et al.; Thirty cases of cutaneous Kaposi's sarcoma (KS) were evaluated and compared with eight cases of acquired immunodeficiency syndrome (AIDS)-related bacillary angiomatosis (BA) . The morphologic features of both lesions were studied by light and electron microscopy and by immunohistochemistry with monoclonal endothelial antibodies against CD34, BNH9, and factor VIII-related antigen as well as the lectins Ulex europaeus 1 and Psophocarpus tetragonolobus . Macrophage/monocyte markers used were alpha 1-antitrypsin, lysosome, Kp1 (CD68), and polyclonal factor XIIIa . Electron microscopic studies demonstrated that most of the spindle cells in KS showed a paucity of cell organelles and an absence of Weibel-Palade bodies (WPB), whereas the cells in BA showed activated endothelial cells with WPB . By immunohistochemistry the spindle cells in KS were consistently positive for CD34 only, whereas proliferating cells in BA expressed all endothelial markers used . Numerous cells expressing macrophage/monocyte markers were present surrounding both KS and BA, and a small number of similar cells were entrapped within both lesions . The results demonstrated a restricted immunohistochemical profile for endothelial cell markers in spindle cells of KS (CD34+) distinct from that of endothelial cells in BA . These findings suggest that the spindle cells in KS are poorly differentiated endothelial cells or that they belong to an endothelial cell subset with partial expression of endothelial phenotype. Clin Infect Dis, 1992 Nov, 15(5), 855 - 7 Significant infections due to Bacillus species following abrasions associated with motor vehicle-related trauma; Wong MT et al.; Non-anthracis Bacillus species are ubiquitous gram-positive spore-forming organisms that were once believed to be nonpathogenic but are now recognized as causing a variety of infections . We report a new aspect of trauma associated with bacillus infection: clinically significant infection by Bacillus species in patients who are involved in motor vehicle accidents and sustain injury related to road contact . Cases were evaluated retrospectively from May 1990 through December 1991 . Four patients who had documented infections with Bacillus species and who were involved in motor vehicle accidents associated with road trauma were identified during this period . The antibiotic susceptibility profile of the Bacillus species consistently demonstrated resistance to beta-lactam antibiotics . This series of cases illustrates an additional aspect of disease associated with bacteria of the Bacillus species that should be considered for patients who have sustained injuries from motor vehicle accidents associated with road trauma. Biochem J, 1992 Nov 1, 287 ( Pt 3), 971 - 7 Purification and properties of DNA polymerase from Bacillus caldotenax; Burrows JA et al.; A thermostable DNA polymerase was prepared from Bacillus caldotenax by using a four-step chromatography procedure . The protein exists as a monomer of M(r) 94,000, has a pI of 4.9 and has no associated 3'-5' or 5'-3'-exonuclease activities or endonuclease activity . The temperature optimum of the enzyme was about 70 degrees C and the pH for maximum activity was about 7.5 . The enzyme has an absolute requirement for a bivalent cation, and maximum activity was obtained at the unusually high concentration of 70 mM-MgCl2 . Mg2+ could be replaced by MnCl2 or CoCl2, with decreased activity, at the lower optimal concentrations of 1 mM and 2.5 mM respectively . Enzyme activity was inhibited in the presence of 2',3'-dideoxy-TTP, arabinosyl-CTP and aphidicolin . Enzyme activity was stimulated with KCl concentrations of about 100 mM, and concentrations of univalent salts above about 150 mM inhibited activity . The enzyme could use activated calf thymus DNA, poly(dA).p(dT)10 or primed single-stranded phage M13 DNA as a template and maximum activity was obtained with poly(dA).p(dT)10 . The enzyme was inactive on unprimed single-stranded DNA, double-stranded DNA and polyribonucleotide template/primer . The apparent Km values for individual dNTPs, determined with the other dNTPs at saturating concentrations, were 5.7 microM (dCTP), 6.3 microM (dATP, dGTP) and 6.4 microM (dTTP) . The Km value for the overall incorporation of each dNTP from an equimolar mixture of all four dNTPs was 24.7 microM . The kcat . value was about 1.05 s-1 . The kcat./Km value was 0.16-0.18 M-1.s-1 for individual dNTPs and 0.04 for the incorporation of an equimolar mixture of all four dNTPs . Some of the properties of the enzyme show it may be classified as an alpha-Type DNA polymerase. Biochem J, 1992 Nov 1, 287 ( Pt 3), 685 - 90 Investigation of the first step of biotin biosynthesis in Bacillus sphaericus . Purification and characterization of the pimeloyl-CoA synthase, and uptake of pimelate; Ploux O et al.; The pimeloyl-CoA synthase from Bacillus sphaericus has been purified to homogeneity from an overproducing strain of Escherichia coli . The purification yielded milligram quantities of the synthase with a specific activity of 1 unit/mg of protein . Analysis of the products showed that this enzyme catalysed the transformation of pimelate into pimeloyl-CoA with concomitant hydrolysis of ATP to AMP . Using a continuous spectrophotometric assay, we have examined the catalytic properties of the pure enzyme . The pH profile under Vmax . conditions showed a maximum around 8.5 . Apparent Km values for pimelate, CoASH, ATP.Mg2- and Mg2+ were respectively 145 microM, 33 microM, 170 microM and 2.3 mM . The enzyme was inhibited by Mg2+ above 10 mM . This acid-CoA ligase exhibited a very sharp substrate specificity, e.g . neither GTP nor pimelate analogues (di- or mono-carboxylic acids) were processed . The bivalent metal ion requirement was also investigated: Mn2+ (73%) and Co2+ (32%) but not Ca2+ could replace Mg2+ . The enzyme was inhibited by metal chelators such as 1,10-phenanthroline and EDTA . The synthase was a homodimer with a 28,000-M(r) subunit . N-Terminal sequencing definitely proved that this enzyme was encoded by the bioW gene . A careful study of pimelate uptake by B . sphaericus, E . coli and Pseudomonas dentrificans showed that this metabolite crossed the membrane of these microorganisms by passive diffusion, ruling out the involvement of the bioX gene product as pimelate carrier. Am Rev Respir Dis, 1992 Nov, 146(5 Pt 1), 1330 - 3 Miliary Mycobacterium bovis induced by intravesical bacille Calmette-Guérin immunotherapy; McParland C et al.; Intravesical instillation of bacille Calmette-Guerin (BCG), an attenuated strain of Mycobacterium bovis, is the treatment of choice for many patients with bladder cancer . In a small percentage, this therapy is associated with systemic side effects including pneumonitis . It is uncertain whether these systemic manifestations are due to dissemination of infection or due to hypersensitivity, an etiologic distinction that has important therapeutic implications . We report the first case in which miliary M . bovis was proven to be the responsible mechanism, by culture of M . bovis biovar BCG from a transbronchial lung biopsy and complete resolution on anti-tuberculous chemotherapy. J Urol, 1992 Nov, 148(5), 1583 - 6 Expression of adhesion molecules by bladder cancer cells: modulation by interferon-gamma and tumour necrosis factor-alpha; Jackson AM et al.; The constitutive expression by eight human bladder cancer cell lines of the cell adhesion molecules intercellular adhesion molecule-1 and intercellular adhesion molecule-2 was studied using monoclonal antibody probes in conjunction with flow-cytometry . Tumour lines of low grade (G1) did not constitutively express intercellular adhesion molecule-1, rather they were found to express intercellular adhesion molecule-2 . The G2 cells expressed no intercellular adhesion molecule-2, however, a low percentage did express intercellular adhesion molecule-1 . High grade cells (G3) only expressed intercellular adhesion molecule-1 on their cell surface but at higher levels than the G2 cell line . Exposure of the bladder cancer cell lines to interferon-gamma induced and augmented the expression of intercellular adhesion molecule-1 by all except one of the cell lines (UMUC3) . Intercellular adhesion molecule-2 expression remained unaltered . The modulation of intercellular adhesion molecule-1 expression was dependent on the concentration of interferon-gamma and the duration of stimulus . De novo intercellular adhesion molecule-1 expression, induced by interferon-gamma, was rapid (< 4 hours) with only a short period of stimulation being required (< 10 seconds) . The rapid increase in expression of intercellular adhesion molecule-1 required de novo protein synthesis and was not the result of release of intercellular adhesion molecule-1 from an intracellular pool . Interferon-gamma and tumour necrosis factor-alpha were found to act synergistically in the induction and augmentation of intercellular adhesion molecule-1 expression . Optimal induction occurred with 10 Uml-1 of both molecules . These results suggest a correlation between constitutive adhesion molecule expression and the histopathological grade of the tumour . The implications of these findings for Bacillus Calmette Guerin and interferon-gamma immunotherapy of bladder cancer is discussed. J Urol, 1992 Nov, 148(5), 1534 - 5 Epididymo-orchitis developing as a late manifestation of intravesical bacillus Calmette-Guerin therapy and masquerading as a primary testicular malignancy: a report of 2 cases; Truelson T et al.; In 2 cases epididymo-orchitis, indistinguishable from a testicular tumor, developed as a late (15 and 34 months, respectively) complication following use of Tice strain bacillus Calmette-Guerin for treatment of superficial bladder carcinoma . In each instance the lesion was asymptomatic and ultrasonography demonstrated a complex scrotal mass . Inguinal orchiectomy was performed for diagnosis and therapy . The importance of obtaining a mycobacterial culture for further therapy planning is stressed. J Lab Clin Med, 1992 Nov, 120(5), 740 - 5 Respiratory epithelial carbohydrate levels of rats with gram-negative bacillary colonization; Mason CM et al.; One mechanism by which severe illness or stress might facilitate adherence and colonization of GNB to respiratory epithelium is by altering epithelial cell surface carbohydrates . To investigate this possibility we used radiolabeled lectins to quantitate carbohydrate levels on intact buccal and tracheal epithelium . A rat model of GNB colonization, in which renal infarction was performed to produce colonization, was used . Buccal and tracheal epithelial surface carbohydrate levels from normal rats and rats 48 hours after renal infarction were compared . Buccal and tracheal epithelium from the renal infarction animals had decreased amounts of sialic acid and fucose, and decreased levels of these sugars occurred at the same time that heavy oropharyngeal GNB colonization developed . Tracheas obtained from the infarcted animals bound three times more Type 1 piliated GNB than normal tracheas . Sialic acid and fucose levels are decreased early after stress, and we speculate that altered epithelial carbohydrates may predispose to GNB colonization by exposing binding sites for GNB. J Histochem Cytochem, 1992 Nov, 40(11), 1779 - 88 Nature and distribution of mineral-binding, keratan sulfate-containing glycoconjugates in rat and rabbit bone; Maeno M et al.; The presence of keratan sulfate (KS) and KS proteoglycans in bone has been demonstrated in birds and rabbits but comparison with other animal species has not been investigated . The nature and distribution of mineral-binding, KS-containing glycoconjugates in rat and rabbit bone were investigated with a monoclonal antibody (MAb 5D4) specific for KS . Mineral-binding proteins were extracted from the mineralized bone with 0.4 M EDTA without guanidine-HCl (E-extract) . On Western blot analysis of SDS-polyacrylamide gel electrophoresis, rat E-extract gave a weak 5D4-reactive band, M(r) 66,000-68,000, whereas rabbit E-extract produced two major reactive populations of small and large molecular size; one population consisted of two closely spaced bands at M(r) 61,000-63,000 and 66,000-68,000, and the other population consisted of one band at approximately M(r) 200,000 . The identity of KS chains was further established by the sensitivity of these bands to keratanase II (Bacillus sp . Ks 36) and endo-beta-galactosidase . Immunocytochemistry with MAb 5D4 showed that, in rat bone, staining associated with the mineral phase was limited to the walls of osteocytic lacunae and bone canaliculi, whereas the remainder of the mineralized matrix lacked staining . In contrast, in rabbit bone the staining was distributed over the entire portion of the mineralized matrix with focal accumulation of staining in the wall of the lacunocanalicular system . These results indicate that rat bone contains a mineral-binding, KS-containing glycoconjugate with preferential localization in the wall of the lacunocanalicular system, whereas rabbit bone contains at least two or possibly three types of KS-containing glycoconjugates distributed over the entire portion of the mineralized matrix. J Bacteriol, 1992 Nov, 174(22), 7478 - 81 Analysis of mutations in cyclodextrin glucanotransferase from Bacillus stearothermophilus which affect cyclization characteristics and thermostability; Fujiwara S et al.; Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) produces cyclodextrin from starch . The CGTase molecule is composed of four globular domains, A, B, C, and D . In order to gain better understanding of the amylolytic and cyclization mechanisms of CGTase, mutant CGTases were constructed from a CGTase gene (cgt1) of Bacillus stearothermophilus NO2 . Cgt1-F191Y (Phe at position 191 was replaced by Tyr), Cgt1-F191Y-F255Y, Cgt1-W254V-F255I, Cgt1-W254V, and Cgt1-F255I were constructed for the analysis of the NH2-terminal region . It was revealed that amino acids surrounding a spiral amylose are important for cyclization characteristics and that hydrophobic amino acids just after the Glu catalytic site play an important role in the hydrolysis characteristics of the enzyme . Mutant CGTases Cgt1-T591F and Cgt1-W629F were also constructed to study the role of a second substrate-binding site in domain D, and it was suggested that substrate binding at both domains A and D stabilized the enzyme and optimized cyclodextrin production. Infect Immun, 1992 Nov, 60(11), 4452 - 9 Mycobacterium leprae produces extracellular homologs of the antigen 85 complex; Pessolani MC et al.; The antigen 85 complex is a set of at least three closely related secreted proteins (85A, 85B, and 85C) of 30 to 32 kDa produced by Mycobacterium tuberculosis and other mycobacteria . Their prominence in Mycobacterium leprae, the one obligate intracellular pathogen of the genus, had been assumed on the basis of immunological evidence and proof of the existence of the gene encoding the 85B protein of the complex . We have now observed the production of this family of proteins by M . leprae through analysis of various fractions by Western blotting (immunoblotting) with monospecific rabbit antisera raised against the individual Mycobacterium bovis BCG 85A, 85B, and 85C proteins . A predominant cross-reactive band with an apparent molecular mass of 30 kDa was detected in extracts of nondisrupted whole M . leprae and in soluble fractions prepared from the tissues of M . leprae-infected armadillos . Further studies of the subcellular distribution of this protein within the bacterium confirmed that it is secreted by the organism, an observation that explains past difficulties in detecting the antigen 85 complex in M . leprae . Confirmation that the M . leprae product is a member of the antigen 85 complex was obtained by comparison of peptide fingerprints with those from the BCG product . The pattern of reactivity of the M . leprae antigen 85 complex with anti-M . bovis BCG 85B serum, as well as two-dimensional electrophoresis, established that the 85B component was the predominant member of the complex in M . leprae . The fibronectin-binding capacity of the M . leprae and BCG 85 complexes was reinvestigated by new approaches and is questioned . Nevertheless, the results obtained with the native proteins reinforce previous reports, derived primarily from the use of homologous proteins, that the antigen 85 complex is one of the dominant protein immunogens of the leprosy bacillus. Appl Microbiol Biotechnol, 1992 Nov, 38(2), 243 - 7 Cloning and expression of a thermostable exo-alpha-1,4-glucosidase gene from Bacillus stearothermophilus ATCC12016 in Escherichia coli; Takii Y et al.; The gene coding for a thermostable exo-alpha-1,4-glucosidase (alpha-glucoside glucohydrolase: EC 3.2.1.20) of Bacillus stearothermophilus ATCC 12016 was cloned within a 2.8-kb AvaI fragment of DNA using the plasmid pUC19 as a vector and Escherichia coli JM109 as a host . E . coli with the hybrid plasmid accumulated exo-alpha-1,4-glucosidase mainly in the cytoplasm . The level of enzyme production was about sevenfold higher than that observed for B . stearothermophilus . The cloned enzyme coincided absolutely with the B . stearothermophilus enzyme in its relative molecular mass (62,000), isoelectric point (5.0), amino-terminal sequence of 15 residues (Met-Lys-Lys-Thr-Trp-Trp-Lys-Glu-Gly-Val-Ala-Tyr-Gln-Ile-Tyr-), the temperature dependency of its activity and stability, and its antigenic determinants. Appl Microbiol Biotechnol, 1992 Nov, 38(2), 147 - 51 Introduction of sulphhydryl groups into the crystalline bacterial cell surface layer protein from Bacillus stearothermophilus PV72 and its application as an immobilization matrix; Sara M et al.; The crystalline cell surface layer (S-layer) from Bacillus stearothermophilus PV72 was used as a matrix for reversible immobilization of beta-D-galactosidase via disulphide bonds . In order to obtain an immobilization matrix stable towards acid, alkali and reducing agents such as dithiothreitol (DTT), the S-layer subunits were first cross-linked with glutaraldehyde . This was done in a way whereby 75% of the free amino groups remained unmodified, and then could be completely converted into sulphhydryl groups upon reaction with the monofunctional imidoester iminothiolane . After activation of the sulphhydryl groups with 2,2'-dipyridyldisulphide, 550 micrograms beta-D-galactosidase could be immobilized per milligram of S-layer protein, which corresponds to one beta-D-galactosidase molecule {relative molecular mass (M(r)), 116,000} per two S-layer subunits (M(r), 130,000) . At least 90% of the sulphhydryl groups from the S-layer protein could be regenerated for further activation by cleaving the disulphide bonds with DTT . In comparative studies beta-D-galactosidase was linked to carbodiimide-activated carboxyl groups of the S-layer protein. Biosci Biotechnol Biochem, 1992 Nov, 56(11), 1792 - 6 Purification and some properties of a Haim-sensitive alpha-amylase from newly isolated Bacillus sp . No . 195; Kawaguchi T et al.; Newly isolated Bacillus sp . No . 195 produced an extracellular alpha-amylase sensitive to Haim which was found to inhibit specifically animal alpha-amylases . The enzyme was purified easily by two steps of starch adsorption and gel filtration using Sephacryl S-200 . The purified enzyme, which showed a single band on native-PAGE or SDS-PAGE, had a molecular weight of 60,000 as judged on SDS-PAGE . The optimum pH value for activity and the isoelectric point were around 7.0 and 4.5, respectively . The sensitivity of the amylase to Haim was similar to that of animal amylase rather than bacterial amylase . It was suggested that a Haim-amylase complex might be formed at the molar ratio of 1:1 . The amino acid sequence F-S-W similar to the triplet F-E-W highly conserved among alpha-amylases sensitive to proteinaceous inhibitors, such as Hoe 467-A or Haim, was found in the amino-terminal part of the No . 195 amylase. Biull Eksp Biol Med, 1992 Nov, 114(11), 510 - 2 {The vitamin D endocrine system and bone tissue mineral metabolism in rats with adjuvant arthritis: the effect of 1,25-dihydroxyvitamin D3}; Sergeev IN et al.; Adjuvant arthritis was induced in male rats by injecting bacillus Calmette-Guerin in mineral oil in a hindpaw . A decrease in bone density, calcium and phosphorus content due to polyarthritis was found in the tibia of the noninjected hind leg . Arthritic rats demonstrated serum 1,25-dihydroxyvitamin D deficiency along with constant level of 25-hydroxyvitamin D . The disease caused a significant expression of 1,25-dihydroxyvitamin D3 receptors in lymphocytes . Arthritic rats were treated with 1,25-dihydroxyvitamin D3 (0.15 mg/kg/day orally) for 35 days . The treatment prevented the development of osteoporosis and a decrease of 1,25-dihydroxyvitamin D levels as well as reduced the expression of 1,25-dihydroxyvitamin D receptors in lymphocytes. Nippon Rai Gakkai Zasshi, 1992 Nov, 61(3), 165 - 74 Demonstration of PGL-I & LAM-B antigens in paraffin sections of leprosy skin lesions; Wang T et al.; An investigation on the demonstration of PGL-I and LAM-B antigens in thirty-four paraffin embedded skin biopsies taken from leprosy patients who covered the whole spectrum of the disease and in four control specimens was carried out . Neither the PGL-I antigen nor the LAM-B antigen was demonstrated in the normal skin specimens that were used as negative control; and only the LAM-B antigen appeared in the tuberculosis specimens in which the PGL-I antigen was negative . The PGL-I antigen was demonstrated on thirty-three leprosy samples except one TT sample and the LAM-B antigen, on all samples by immunochemical staining technique . The antigens were identified as intracytoplasmic bacillary staining, in solitary, granular as well as debris patterns; and as soluble antigenic staining, in vacuolar or amorphous pattern . In LL and BL cases, the antigens were detected predominantly from macrophages and peripheral nerves in all five staining patterns; in BB cases, from macrophages mostly in the granular as well as debris patterns, from the nerves in the vacuolar pattern; while in TT and the majority of BT cases, they were mainly from nerve remnants inside the granuloma in the vacuolar or amorphous staining pattern . In addition, it is interesting to note that the immunochemical staining was able to differentiate the foamy change from the hydropic degeneration . We also found that the antigens distributed in arrector pili muscles and the walls of muscular vessels were obviously related to the unmyelinated nerve fibers innervating the smooth muscle cells.(ABSTRACT TRUNCATED AT 250 WORDS) Eur J Clin Microbiol Infect Dis, 1992 Nov, 11(11), 1058 - 63 Clinical importance of Bilophila wadsworthia; Finegold S et al.; Bilophila wadsworthia is an anaerobic, gram-negative, asaccharolytic, urease-positive, bile-resistant, catalase-positive bacillus, originally recovered from infections in patients with gangrenous and perforated appendicitis . Additional isolations from clinical specimens, including pleural fluid, joint fluid, blood and pus from a scrotal abscess, mandibular osteomyelitis and axillary hidradenitis suppurativa are described here . Bilophila is found as normal flora in feces and, occasionally, in saliva and in the vagina . Isolates from humans are usually beta-lactamase positive and therefore resistant to certain beta-lactam antibiotics . Two percent of strains are also resistant to clindamycin. Trans R Soc Trop Med Hyg, 1992 Nov-Dec, 86(6), 686 - 92 Does antibody to mycobacterial antigens, including lipoarabinomannan, limit dissemination in childhood tuberculosis? Costello AM, Kumar A, Narayan V, Akbar MS, Ahmed S, Abou-Zeid C, Rook GA, Stanford J, Moreno C. Serum immunoglobulin (Ig) G responses to a variety of mycobacterial antigens were measured in children from the UK, in children with tuberculosis from Hyderabad, India and Dhaka, Bangladesh, classified according to whether the disease was disseminated or localized, and in non-tuberculous controls . Anti-lipoarabinomannan (LAM) IgG responses in UK children showed a marked trough between 6 months and 3 years coincident with the reported peak incidence of disseminated tuberculosis . Geometric mean IgG responses to sonicates of slow-growing mycobacteria (rich in LAM) in 36 children with disseminated tuberculosis were markedly lower than in 99 children with localized tuberculous lesions (for Mycobacterium scrofulaceum P < 0.01, for M . tuberculosis P < 0.01, and for M . vaccae P < 0.01) . Responses to purified LAM were also lower in the disseminated tuberculosis group (P < 0.05) but there was no difference between the groups in their response to mycobacterial 65 kDa protein . Multiple regression analysis showed that the reduced response to sonicated mycobacterial antigens and to LAM in children with disseminated disease was independent of age, nutritional status, skin test reactivity, duration of previous symptoms, and city of origin . There was no evidence for sequestration of antibody to immune complexes . These findings are compatible with the hypothesis that children with low levels of antibody to sonicated mycobacterial antigen and to LAM, or those who cannot mount an antibody response, are predisposed to dissemination . A role for antibody in preventing disseminated forms of tuberculosis in childhood has implications for the development of improved vaccines and for the optimum timing of vaccination with bacille Calmette-Guerin. Plant Mol Biol, 1992 Nov, 20(3), 539 - 48 Expression of a chimeric CaMV 35S Bacillus thuringiensis insecticidal protein gene in transgenic tobacco; Carozzi NB et al.; Insecticidal transgenic tobacco plants containing a truncated Bacillus thuringiensis cryIA(b) crystal protein (ICP) gene expressed from the CaMV 35S promoter were analyzed for ICP gene expression under field and greenhouse conditions over the course of a growing season . We present new information on temporal and tissue-specific expression of a CaMV 35S/cryIA(b) gene . Levels of cryIA(b) protein and mRNA were compared in both homozygous and hemizygous lines throughout plant development . Levels of ICP mRNA and protein increased during plant development with a pronounced rise in expression at the time of flowering . Homozygous ICP lines produced higher levels of ICP than the corresponding hemizygous lines . ELISA analysis of different tissues in the tobacco plant showed ICP gene expression in most tissues with a predominance of ICP in older tissue . All transgenic ICP tobacco lines which were studied in the field and greenhouse contained 400 ng to 1 microgram ICP per gram fresh weight in leaves from the mid-section of the plant at flowering . The amounts of ICP produced by field lines were directly comparable to levels observed in greenhouse-grown plants. Appl Microbiol Biotechnol, 1992 Nov, 38(2), 173 - 8 Isolation of a gramicidin S hyperproducing strain of Bacillus brevis by use of a fluorescence activated cell sorting system; Azuma T et al.; A gramicidin S (GS) hyperproducing mutant of Bacillus brevis was isolated by using a protein-staining fluorescence dye (fluorescein isothiocyanate, FITC), and a fluorescence-activated cell sorting system (FACS) . By flow cytometry (FCM) analysis after staining with FITC, higher producing cells of the wild-type had higher fluorescence signals than cells with low productivity or cells from a GS non-producing mutant . Staining with FITC did not affect the viability of cells under the conditions chosen for FCM analysis . This enabled us to recover viable cells after sorting . After wild-type cells were mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine, mutants with higher fluorescence than the parental strain were obtained by cell sorting . Among them, strain 18 was chosen as a GS hyperproducer; it produced 590 micrograms GS/ml compared to 350 micrograms/ml by the wild-type strain . This method has the advantage of being able to screen large numbers of cells in a short time . Furthermore, use of the fluorescence dye technique will expand the use of FACS to the improvement of other cultures that produce metabolites that do not have a specific fluorescence or strong enough fluorescence for normal cell sorting. J Mol Biol, 1992 Oct 20, 227(4), 1263 - 4 Purification, crystallization and preliminary X-ray analysis of the 3-phosphoglycerate kinase from Bacillus stearothermophilus; Davies GJ et al.; As part of a programme investigating the molecular basis of thermal stability in proteins we have isolated and characterized the thermally stable 3-phosphoglycerate kinase (PGK) from Bacillus stearothermophilus NCA 1503 . The B . stearothermophilus PGK has been crystallized in a form suitable for X-ray diffraction analysis . Crystals which diffract to greater than 1.8 A resolution have been grown in the presence of the nucleotide substrate, MgATP, using polyethylene glycol (PEG 600) as a precipitant . The best crystals have been obtained using "seeding" techniques and are monoclinic, space group P2(1), with cell dimensions a = 40.5 A, b = 74.0 A, c = 68.5 A and beta = 99.8 degrees. Biochim Biophys Acta, 1992 Oct 19, 1111(1), 27 - 34 Vasopressin stimulation of vanadate-sensitive Na+ transport by liver plasma membrane vesicles . Evidence for regulation via phospholipase C and protein kinase C activities; Piec G et al.; The rate of vanadate-sensitive 22Na+ uptake by isolated liver membrane vesicles, reflecting transport by Na+/K(+)-ATPase, was measured to study the role played by phospholipase C and protein kinase C in the regulation of this process by vasopressin . Na+ uptake was enhanced 2-3-fold by 100 nM {Arg8}vasopressin and the hormone effect was mimicked by 0.1 microM inositol 1,4,5-trisphosphate as well as by 1.0 microM myo-inositol . The stimulation by vasopressin was potentiated by phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis (5-10 mU/ml) . No effect of the bacterial enzyme was observed in the absence of the hormone . Phorbol myristate acetate (0.5-1 microM) suppressed the stimulation by vasopressin but had no effect in the absence of the hormone . High concentrations of bacterial phosphatidylinositol-specific phospholipase C (50-100 mU/ml) also antagonized the hormone stimulation . Staurosporine (50-100 nM) prevented the antagonistic effect of bacterial phospholipase C (50 mU/ml) and EGTA (1 mM) partially protected the hormonal stimulation in the presence of phorbol myristate acetate . Our results suggest that the stimulatory effect of vasopressin on Na+ transport is mediated by phospholipase C and products derived from the inositol moiety of membrane phospholipids . Membrane-associated protein kinase C appears to be at least partially responsible for the desensitization to stimulation by vasopressin. Presse Med, 1992 Oct 17, 21(34), 1625 - 30 {Bacillary angiomatosis}; Robert C et al.; Bacillary angiomatosis (BA) is a recently described infection usually found in patients with human immunodeficiency virus disease . BA is caused by a Gram-negative coccobacillus . This organism is primarily responsible for skin lesions of the pseudo-botryomycoma type or inflammatory nodules, but it also produces fever, degradation of the general condition and visceral lesions involving the lymph nodes, the liver, the spleen and the bones . Histology shows vascular proliferation with turgid endothelial cells and mostly neutrophilic inflammatory infiltrates . BA is susceptible to many antibiotics . The authors describe the history of the disease and its clinical and histological features, discuss its differential diagnosis and principally deal with the relationship between BA and cat-scratch disease and between BA and verruca peruana . They also present the molecular biology technique which enables a genotypic diagnosis of the disease to be made, replacing a deficient phenotype. Cancer Res, 1992 Oct 15, 52(20), 5663 - 7 Immunization of colorectal cancer patients with modified ovine submaxillary gland mucin and adjuvants induces IgM and IgG antibodies to sialylated Tn; O'Boyle KP et al.; Tn and sialylated Tn (sTn) are blood group-related epitopes expressed on mucins of colon carcinoma and other epithelial tumors and are, therefore, potential targets for immunological control . We have immunized 20 colorectal cancer patients at high risk for recurrence with a vaccine consisting of partially desialylated ovine submaxillary gland mucin (modified OSM) which contains both Tn and sTn determinants . Six patients were treated with modified OSM alone (group 1), eight patients were treated with modified OSM and the immunological adjuvant DETOX (group 2), and six patients were treated with modified OSM and Bacillus Calmette-Guerin (group 3) . Pre- and postvaccination sera were tested by enzyme-linked immunosorbent assay and dot blot immune stains for antibodies reactive with modified OSM . Antibody titers increased in 4 of 8 patients immunized with modified OSM and DETOX, in 5 of 6 patients immunized with modified OSM and B . Calmette-Guerin, and in 0 of 6 patients receiving modified OSM without adjuvant . The specificity of induced IgM and IgG antibodies was confirmed by demonstrating reactivity with OSM, bovine submaxillary mucin, and synthetic glycoconjugates sTn-human serum albumin (HSA) and Tn-HSA in enzyme-linked immunosorbent assay and immune stains . Median IgM pre-postvaccination reciprocal titers were 20/80 for Tn-HSA and 10/320 for sTn-HSA . Low level IgG antibody titers against sTn-HSA were detected after vaccination in 7 patients . Toxicity was limited to inflammatory skin reactions at the site of vaccination resulting from the adjuvants . No inflammatory infiltrates were seen in the skin when the modified OSM vaccine was administered in the absence of an immunological adjuvant . These results demonstrate that sTn and Tn can be recognized by the human immune system and that vaccines containing these structures can be administered safely with immunological adjuvants . Attempts to augment the immunogenicity of these carbohydrate antigens by covalent attachment to immunogenic carrier proteins and the use of more potent immunological adjuvants are now being pursued. J Biol Chem, 1992 Oct 5, 267(28), 19919 - 23 Mammalian cells that express Bacillus cereus phosphatidylinositol-specific phospholipase C have increased levels of inositol cyclic 1:2-phosphate, inositol 1-phosphate, and inositol 2-phosphate; Ross TS et al.; Phosphatidylinositol-specific phospholipase C (PtdIns-PLC) of Bacillus cereus catalyzes the conversion of PtdIns to inositol cyclic 1:2-phosphate and diacylglycerol . NIH 3T3, Swiss mouse 3T3, CV-1, and Cos-7 cells were transfected with a cDNA encoding this enzyme, and the metabolic and cellular consequences were investigated . Overexpression of PtdIns-PLC enzyme activity was associated with elevated levels of inositol cyclic 1:2-phosphate (2.5-70-fold), inositol 1-phosphate (2-20-fold), and inositol 2-phosphate (3-20-fold) . The increases correlated with the levels of enzyme expression obtained in each cell type . The turnover of phosphatidylinositol (PtdIns) was also increased in transfected CV-1 cells by 13-fold 20 h after transfection . The levels of PtdIns, phosphatidic acid, diacylglycerol, or other inositol phosphates were not detectably altered . Expression of bacterial PtdIns-PLC decreased rapidly after 20 h implying that either the increased PtdIns turnover or the accumulation of inositol phosphates was detrimental to cells and that by some adaptive mechanism enzyme expression was suppressed. J Infect Dis, 1992 Oct, 166(4), 874 - 84 The 38-kDa protein of Mycobacterium tuberculosis: a review; Harboe M et al.; This review illustrates that the 38-kDa protein is one of the most important antigens of Mycobacterium tuberculosis . It is actively secreted but partly attached to the surface of the mycobacterial cell by a lipid tail that may also be responsible for binding of carbohydrate to the protein . It is a major constituent of M . tuberculosis culture fluid after growth on the synthetic Sauton medium and occurs in bacille Calmette-Guerin in far lower concentrations . The protein induces B and T cell responses with high specificity for infection with M . tuberculosis and is a prime candidate for development of new diagnostic reagents for tuberculosis. Plant Mol Biol, 1992 Oct, 20(1), 81 - 93 Arabidopsis thaliana small subunit leader and transit peptide enhance the expression of Bacillus thuringiensis proteins in transgenic plants; Wong EY et al.; The expression of the modified gene for a truncated form of the cryIA(c) gene, encoding the insecticidal portion of the lepidopteran-active CryIA(c) protein from Bacillus thuringiensis var . kurstaki (B.t.k.) HD73, under control of the Arabidopsis thaliana ribulose-1,5-bisphosphate carboxylase (Rubisco) small subunit ats1A promoter with and without its associated transit peptide was analyzed in transgenic tobacco plants . Examination of leaf tissue revealed that the ats1A promoter with its transit peptide sequence fused to the truncated CryIA(c) protein provided a 10-fold to 20-fold increase in cryIA(c) mRNA and protein levels compared to gene constructs in which the cauliflower mosaic virus 35S promoter with a duplication of the enhancer region (CaMV-En35S) was used to express the same cryIA(c) gene . Transient expression assays in tobacco protoplasts and the whole plant results support the conclusion that the transit peptide plus untranslated sequences upstream of that region are both required for the increase in expression of the CryIA(c) protein . Furthermore, the CaMV-En35S promoter can be used with the Arabidopsis ats1A untranslated leader and transit peptide to increase expression of this protein . While subcellular fractionation revealed that the truncated CryIA(c) protein fused to the ats1A transit peptide is located in the chloroplast, the increase in gene expression is independent of targeting of the CryIA(c) protein to the chloroplast . The results reported here provide new insight into the role of 5' untranslated leader sequences and translational fusions to increase heterologous gene expression, and they demonstrate the utility of this approach in the development of insect-resistant crops. J Biochem (Tokyo), 1992 Oct, 112(4), 488 - 91 Mass production of sphingomyelinase of Bacillus cereus by a protein-hyperproducing strain, Bacillus brevis 47, and its purification; Tamura H et al.; Sphingomyelinase (sphingomyelin cholinephosphohydrolase) {EC 3.1.4.12} of Bacillus cereus was overproduced in a protein-hyperproducing strain, B . brevis 47, by cloning the gene into an expression vector pNU211, which has been developed to express a foreign gene utilizing a promoter and a signal sequence of an outer cell wall protein gene . From 1 liter of culture, about 10 mg of protein was purified to near-homogeneity by two steps of column chromatography; this is almost 500 times higher production compared to the conventional preparation from the original strain, B . cereus IAM 1208 . The N-terminal amino acid sequence of the secreted enzyme was identical to that of the authentic enzyme, indicating that the signal sequence for secretion of B . cereus was processed properly in B . brevis 47. Int J Food Microbiol, 1992 Oct, 17(2), 85 - 99 Media for the detection and enumeration of Bacillus cereus in foods: a review; van Netten P et al.; Bacillus cereus is an established cause of food poisoning in addition to being a troublesome and persistent contaminant, responsible for a variety of spoilage defects in processed foods and dairy products . A range of diagnostic and selective media has been developed to facilitate the detection and enumeration of B . cereus in routine surveillance situations and food poisoning investigations . These media are reviewed with respect to the selective and diagnostic systems they employ, their ability to recover and differentiate the target organism, and their advantages and limitations in particular applications. Wei Sheng Wu Xue Bao, 1992 Oct, 32(5), 314 - 9 {Molecular cloning and identification of 130kd mosquitocidal protein gene of Bacillus thuringiensis var . israelensis (Bti)}; Hua X et al.; The location of 130kd mosquitocidal protein gene of Bti 4Q5 strain on its 75Md plasmid was confirmed by southern hybridization using a 18-base oligonucleotide probe . The crystal protein containing the component of 130kd toxic protein was purified . The crystal protein exhibiting the mosquitocidal activity against larvae of Aedes aegypti was shown by bioassay . The purified 75Md plasmid DNA of Bti 4Q5 strain was completely digested with HindIII restriction enzyme, ligated with the vector pUC18 and transformed into the recipient cells of E . coli TG1 . From Apr transformants, four clones with HindIII restriction fragment inserts highly homologous to the 18-base oligonucleotide probe were obtained by in situ hybridization and southern hybridization . The 5.2kb HindIII restriction fragment insert was obtained in clone pFH2 and clone pFH4, and 2.3kb HindIII restriction fragment insert in clone pFH1 and pFH3 . For pFH2 and pFH4, the 5.2kb fragment was inserted in pUC18 in opposite orientation . It contained 130kd mosquitocidal protein gene (type I) identified by restriction enzyme map analysis . The 2.3kb HindIII fragment insert in other two clones (pFH1 and pFH3) harbored a part of the type II mosquitocidal protein gene which can be used as a probe for cloning of the type II mosquitocidal protein gene. Protein Eng, 1992 Oct, 5(7), 693 - 701 Site-saturation mutagenesis and three-dimensional modelling of ROB-1 define a substrate binding role of Ser130 in class A beta-lactamases; Juteau JM et al.; Site-saturation mutagenesis was performed on the class A ROB-1 beta-lactamase at conserved Ser130, which is centrally located in the antibiotic binding site where it can participate in both protein-protein and protein-substrate hydrogen bonding . Mutation Thr130 gave a beta-lactamase hydrolysing penicillins and cephalosporins but which showed a 3-fold lower affinity (Km) for ampicillin and cephalexin, and a 30-fold lower hydrolytic (Vmax) activity for ampicillin . In contrast, the hydrolytic activity for cephalexin was similar to the wild-type for the Thr130 mutation . Mutation Gly130 gave a beta-lactamase hydrolysing only penicillins with an affinity and hydrolysis activity for these compounds approximately 15-fold lower than the wild-type, but no detectable activity against cephalosporins . Mutation Ala130 produced an enzyme capable of hydrolysing penicillins only at a low rate . Modelling the ROB-1 active site was done from the refined 2 A X-ray structure of the homologous Bacillus licheniformis beta-lactamase . Ampicillin and cephalexin were docked into the active site and were energy minimized with the CVFF empirical force field . Dockings were stable only when Ser70 was made anionic and Glu166 was made neutral . Interaction energies and distances were calculated for fully hydrated pre-acylation complexes with the Ser, Thr, Gly and Ala130 enzymes . The catalytic data from all mutations and the computed interactions from modelling confirmed that the Ser130 has a structural as well as a functional role in binding and hydrolysis of penicillins . This highly conserved residue also plays a substrate specificity role by hydrogen binding the carboxylic acid group of cephalosporins more tightly than penicillins. Protein Eng, 1992 Oct, 5(7), 611 - 5 The structural consequences of exchanging tryptophan and tyrosine residues in B . stearothermophilus lactate dehydrogenase; Roper DI et al.; A mutant Bacillus stearothermophilus lactate dehydrogenase has been prepared in which all three tryptophan residues in the wild-type enzyme have been replaced by tyrosines . In addition, a tyrosine residue has been mutated to a tryptophan, which acts as a fluorescence probe to monitor protein folding . The mutant enzyme crystallizes in the same crystal form as the wild-type . The crystal structure of the mutant has been determined at 2.8 A resolution . Solution studies have suggested that there is little effect upon the mutant enzyme as judged by its kinetic properties . Comparison of the crystal structures of the mutant and wild-type enzymes confirms this conclusion, and reveals that alterations in structure in the region of these mutations are of a similar magnitude to those observed throughout the structure, and are not significant when compared with the errors in atomic positions expected for a structure at this resolution. Immunol Cell Biol, 1992 Oct, 70 ( Pt 5), 295 - 300 Antibodies to 65 kDa and 70 kDa heat shock proteins in rheumatoid arthritis and systemic lupus erythematosus; Panchapakesan J et al.; To test the potential role of autoimmunity to the highly conserved heat shock proteins (HSP) in immune arthritides, the sera from 99 patients with rheumatoid arthritis (RA), 48 patients with systemic lupus erythematosus (SLE) and 65 normal controls were examined by ELISA for IgG and IgM antibodies to the 65 kDa and 70 kDa heat shock proteins from Mycobacterium bovis (Bacille Calmette-Guerin; BCG) . In RA sera there are significant numbers of individuals with increased IgM anti-65 kDa and anti-BCG reactivity as well as IgG anti-70 kDa when compared with controls . In SLE both IgM and IgG anti-BCG, together with IgM anti-65 kDa, differed significantly from controls . The results were compared with previous reports in similar groups of patients, and it is clear that no consistent pattern of reactivity emerges . While further work may be justified looking carefully at the disease duration and other subsets of both RA and SLE, it is difficult at this stage to conclude that antibodies to autologous HSP that cross-react with mycobacterial HSP play a major role in disease pathogenesis. Planta Med, 1992 Oct, 58(5), 405 - 9 Effects of Phaeodactylum tricornutum and Dunaliella tertiolecta extracts on the central nervous system; Villar R et al.; Continuing work on the effects of aqueous extracts of the microalgae Phaeodactylum tricornutum Bohlin (Bacillariophyceae) and Dunaliella tertiolecta Butcher (Chlorophyceae) on the central nervous system, we report their effects on spontaneous motor activity, rectal temperature, exploratory behaviour, muscle relaxation, catalepsy, and conditioned avoidance responses . Both extracts showed activity as a CNS depressant and a potential muscle relaxant, the latter more marked in the case of P . tricornutum. Planta Med, 1992 Oct, 58(5), 398 - 404 Effects of Skeletonema costatum extracts on the central nervous system; Villar R et al.; We report the effects of aqueous extracts of the microalga Skeletonema costatum Greve (Cleve) (Bacillariophyceae) on spontaneous motor activity, rectal temperature, motor coordination, amphetamine-induced hypermotility, exploratory behaviour, muscle relaxation, catalepsy, conditioned avoidance responses, oxotremorin-induced cholinergic syndrome, and pentylenetetrazole-induced convulsions . The S . costatum extract at dosages of 310 and 620 mg/kg works like an antidopaminergic drug with anticholinergic properties, that does not induce catalepsy and with a notable muscle relaxing activity. Lab Anim Sci, 1992 Oct, 42(5), 449 - 53 Correlation between megaloileitis and antibodies to Bacillus piliformis in laboratory rat colonies; Hansen AK et al.; Rat colonies in which antibodies to Bacillus piliformis were detected in animals examined at the age of 8 to 15 weeks were compared with rat colonies where no such antibodies were present . The seropositive colonies had a low incidence of megaloileitis in 5-week-old rats of Sprague-Dawley stock and some few inbred strains . In seronegative colonies, no megaloileitis was detected . In rats with megaloileitis, significantly high titers to B . piliformis were noted and the agents could be identified in the ileal mucosa by immunofluorescence technique. Lab Anim Sci, 1992 Oct, 42(5), 444 - 8 Rederivation of rat colonies seropositive for Bacillus piliformis and the subsequent screening for antibodies; Hansen AK et al.; Latent infection of rats in a breeding colony with Bacillus piliformis detectable by antibodies to the agent in an immunofluorescence assay was eliminated by a combination of traditional rederivation techniques, using animal units not previously used for rat breeding, and the use of specific disinfection procedures . The success rate was apparently correlated with the use of peracetic acid instead of aldehyde products to decontaminate the animal unit. Lab Anim Sci, 1992 Oct, 42(5), 439 - 43 Subclinical infection and transmission of Tyzzer's disease in rats; Motzel SL et al.; Two isolates of Bacillus piliformis originally obtained from rats from Japan and Indiana were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting . Protein and antigen profiles revealed heterogeneity between the two isolates, demonstrating that more than one isolate of B . piliformis is capable of infecting rats . Results of parallel infection and transmission studies with the two isolates were almost identical . Orally inoculated rats remained asymptomatic; however, enzyme-linked immunosorbent assay results revealed a significant increase in serum antibodies to B . piliformis . Formalin-killed B . piliformis elicited no serum antibody response among rats inoculated orally, indicating that viable organisms, capable of replicating within the host, are needed to induce a systemic humoral response . Naive rats and weanling gerbils were housed on soiled bedding from the experimentally infected, asymptomatic, seropositive rats . Although gerbils showed no clinical signs or histopathologic evidence of Tyzzer's disease, rats housed on bedding collected 1 or 2 weeks postinoculation seroconverted and remained seropositive but asymptomatic throughout the study . These results demonstrate that subclinically infected rats are capable of transmitting B . piliformis to naive rats and suggest that the histopathologic evaluation of sentinel gerbils may not be an effective method for detecting all strains of B . piliformis. Biochem Int, 1992 Oct, 28(1), 97 - 103 Purification and properties of 2,3-dihydroxy-p-cumate-3,4-dioxygenase from Bacillus species; Ninnekar HZ; 2,3-Dihydroxy-p-cumate-3,4-dioxygenase, an enzyme involved in the catabolism of p-cymene, was purified to homogeneity from Bacillus species by affinity chromatography . Purification of the dioxygenase allowed the observation of the immediate ring cleavage product of 2,3-dihydroxy-p-cumate . The enzyme was optimally active at pH 8.2 and at 35 degrees C . The Km value for 2,3-dihydroxy-p-cumate was 32 microM . The enzyme had a broad substrate specificity for 3-substituted catechols . The activity of the enzyme was inhibited by heavy metals, sulphydryl inhibitors, iron-chelating agents, and substrate analogues . Fe2+ was suggested as a cofactor. Appl Environ Microbiol, 1992 Oct, 58(10), 3343 - 6 Evaluation of synergism among Bacillus thuringiensis toxins; Tabashnik BE; A simple test for synergism among toxins is described and applied to previously reported data on independent and joint toxicities of insecticidal proteins from Bacillus thuringiensis . The analysis shows synergism between a 27-kDa (CytA) toxin and 130- or 65-kDa (CryIV) toxins from B . thuringiensis subsp . israelensis against Aedes aegypti larvae . No positive synergism between 130- and 65-kDa toxins or among three CryIA toxins tested against seven species of Lepidoptera occurred . Comparisons with the original interpretations of these data show one case in which synergism occurred but was reported previously as absent and two cases that were not synergistic but were reported previously as suggestive of synergism . These results show that lack of an appropriate test for synergism can produce misleading conclusions . The methods described here can be used to test for synergistic effects of any poisons. Mol Gen Genet, 1992 Oct, 235(1), 147 - 52 Cloning and molecular characterization of the secY genes from Bacillus licheniformis and Staphylococcus carnosus: comparative analysis of nine members of the SecY family; Tschauder S et al.; SecY is a central component of the export machinery that mediates the translocation of secretory proteins across the plasma membrane of Escherichia coli . We have cloned and sequenced the secY genes from Bacillus licheniformis and Staphylococcus carnosus . The deduced amino acid sequences are highly homologous to those of other known SecY polypeptides, all having the potential to form 10 transmembrane segments . Comparative analysis of 9 SecY polypeptides, derived from different bacteria, revealed that 14 amino acid positions (2.7%) are identical in all SecY proteins and 89 (16.9%) show conservative changes . Clusters of conserved amino acid residues were found in 4 of the 10 transmembrane segments and 2 of the 6 cytoplasmic domains . It is suggested that the conserved regions might be involved in the translocation activity of SecY or might be required for the correct interaction of SecY with other components of the secretion apparatus. Mikrobiyol Bul, 1992 Oct, 26(4), 355 - 8 {The growth of some Bradyrhizobium strains in uremic conditions in the presence of antimicrobial substances}; Ayhan K et al.; The growth rate of Bradyrhizobium japonicum USDA 110 and its STRR and AMPR mutant strains (resistant to streptomycin and ampicillin respectively), as well as TAL 945 and TAL 946, TAL 947 (mutants of USDA 110 and USDa 138 respectively) on yeast extract mannitol agar (YEMA) containing different concentrations of Crystal Violet and Brilliant Green . According to our results only the growth of Bacillus megaterium B17 was effected from Crystal Violet while the others were not effected from both of dyes in any concentration. Nippon Hinyokika Gakkai Zasshi, 1992 Oct, 83(10), 1689 - 95 {Prophylaxis of superficial bladder tumor with the intravesical instillation of bacillus Calmette-Guerin or anti-cancer agents . A retrospective study}; Suzuki T et al.; It was retrospectively analyzed whether intravesical instillation of bacillus Calmette-Guerin (BCG) or anti-cancer agents had prophylactic effect or not after removal of superficial bladder tumors . The results over a follow-up period ranging from 6 to 40 months showed 23.8 per cent recurrence in group 1 patients treated with BCG (21 patients), 46.7 per cent recurrence in group 2 treated with anti-cancer agents (45 patients) and 52.3 per cent recurrence in the group 3 (127 patients) which were not received any intravesical drugs . Long-term results among the 3 groups calculated with Kaplan-Meier method demonstrated that the instillation of BCG or of anti-cancer agents was more useful for prophylaxis, compared with the actuarial non-recurrence rates of group 3 . The instillation of BCG showed good prophylactic effects especially in recurrent, grade 2 and pTa bladder tumors . The instillation of anti-cancer agents showed to provide prolonged protection from recurrence, but the instillation of BCG did not. Proteins, 1992 Oct, 14(2), 224 - 36 Effects of changing the interaction between subdomains on the thermostability of Bacillus neutral proteases; Eijsink VG et al.; Variants of the thermolabile neutral protease (Npr) of B . subtilis (Npr-sub) and the thermostable neutral protease of B . stearothermophilus (Npr-ste) were produced by means of site-directed mutagenesis and the effects of the mutations on thermostability were determined . Mutations were designed to alter the interaction between the middle and C-terminal subdomain of these enzymes . In all Nprs a cluster of hydrophobic contacts centered around residue 315 contributes to this interaction . In thermostable Nprs (like Npr-ste) a 10 residue beta-hairpin, covering the domain interface, makes an additional contribution . The hydrophobic residue at position 315 was replaced by smaller amino acids . In addition, the beta-hairpin was deleted from Npr-ste and inserted into Npr-sub . The changes in thermostability observed after these mutations confirmed the importance of the hydrophobic cluster and of the beta-hairpin for the structural integrity of Nprs . Combined mutants showed that the effects of individual mutations affecting the interaction between the subdomains were not additive . The effects on thermostability decreased as the strength of the subdomain interaction increased . The results show that once the subdomain interface is sufficiently stabilized, additional stabilizing mutations at the same interface do not further increase thermostability . The results are interpreted on the basis of a model for the thermal inactivation of neutral proteases, in which it is assumed that inactivation results from the occurrence of local unfolding processes that render these enzymes susceptible to autolysis. J Parasitol, 1992 Oct, 78(5), 800 - 4 Tikusnema javaense n . gen., n . sp . (Nematoda: Acuarioidea) and other nematodes from Rattus argentiventer collected in west Java, Indonesia; Hasegawa H et al.; Nematodes collected from the ricefield rat, Rattus argentiventer (Rodentia: Muridae), in Pusakanagara and Sukamandi, West Java, Indonesia, are reported . Tikusnema javaense n . gen., n . sp . (Nematoda:Acuariidae:Seuratiinae) is described from the small intestine . This new genus is distinguished readily from other genera of the subfamily Seuratiinae in having 4 strongly protruded cuticular leaves in the posterior cephalic portion and in having a pair of prominent cuticular ornamentations posterior to deirids . Besides T . javaense, Eucoleus bacillatus, Strongyloides ratti, Nippostrongylus brasiliensis, Syphacia muris, and Physaloptera sp . were detected. J Pediatr, 1992 Oct, 121(4), 574 - 8 Bacillary angiomatosis in a child undergoing chemotherapy; Myers SA et al.; Bacillary angiomatosis is an infectious disease of the skin and viscera characterized by vascular lesions, originally described in patients with human immunodeficiency virus infection . There are also case reports of bacillary angiomatosis occurring in immunocompetent patients and in noninfected patients with suppressed immune function . We report a case of bacillary angiomatosis in a child undergoing chemotherapy for acute leukemia. J Bacteriol, 1992 Oct, 174(19), 6171 - 8 Structure, function, and fate of the BlaR signal transducer involved in induction of beta-lactamase in Bacillus licheniformis; Zhu Y et al.; The membrane-spanning protein BlaR is essential for the induction of beta-lactamase in Bacillus licheniformis . Its nature and location were confirmed by the use of an antiserum specific for its carboxy-terminal penicillin sensor, its function was studied by genetic dissection, and the structure of the penicillin sensor was derived from hydrophobic cluster analysis of the amino acid sequence by using, as a reference, the class A beta-lactamases with known three-dimensional structures . During the first 2 h after the addition of the beta-lactam inducer, full-size BlaR, bound to the plasma membrane, is produced, and then beta-lactamase is produced . By 2 h after induction, BlaR is present in various (membrane-bound and cytosolic) forms, and there is a gradual decrease in beta-lactamase production . The penicillin sensors of BlaR and the class D beta-lactamases show strong similarities in primary structures . They appear to have the same basic spatial disposition of secondary structures as that of the class A beta-lactamases, except that they lack several alpha helices and, therefore, have a partially uncovered five-stranded beta sheet and a more readily accessible active site . Alterations of BlaR affecting conserved secondary structures of the penicillin sensor and specific sites of the transducer annihilate beta-lactamase inducibility. Infect Immun, 1992 Oct, 60(10), 4051 - 8 Adhesion to and invasion of cultured human cells by Bartonella bacilliformis; Hill EM et al.; Bartonella bacilliformis was tested for its ability to adhere to and invade tissue culture cell monolayers . The parasite was able to efficiently bind and penetrate human dermal fibroblasts, human laryngeal epithelium, and human umbilical vein endothelial cells . Exposure of the organism to immune serum prepared against a crude Bartonella extract containing cell wall and membranous material resulted in decreased ability of the parasite to invade host cells . There was also an overall reduction in the invasiveness of bartonellae and total host cell association when human laryngeal epithelial cells and human umbilical vein endothelial cells were preexposed to cytochalasin D, indicating an active involvement of host cells in the uptake of bartonellae . Transmission electron microscopy revealed the presence of bartonellae inside and outside intracellular vacuoles . These data suggest that a surface-associated factor is involved in the invasion process and that internalization of the parasite by host cells involves a microfilament-dependent process similar to phagocytosis. Biosci Biotechnol Biochem, 1992 Oct, 56(10), 1552 - 6 Enzymatic synthesis of 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltopentaoside, a substrate for alpha-amylase; Ishimaru K et al.; A transglycosylation reaction with 2-chloro-4-nitrophenyl beta-maltoside as an acceptor was done with 4,6-O-3-ketobutylidene maltopentaose and Bacillus macerans cyclodextrin glucanotransferase in an aqueous solution containing 50% n-propanol, and there were two main transglycosylation products . They were identified as 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltopentaoside and 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltohexaoside, and their yields were 30% and 21% respectively on the basis of the decrease of 4,6-O-3-ketobutylidene maltopentaose . For the production of 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltopentaoside at high substrates concentrations, the addition of n-propanol in this reaction not only increased the solubility of 2-chloro-4-nitrophenyl beta-maltoside sufficiently but also suppressed side reactions. Indian J Exp Biol, 1992 Oct, 30(10), 915 - 7 Sensitivity of mosquito-pathogenic bacterial strains to various antibodies; Gupta DK et al.; Four strains of Bacillus sphaericus, 1593, 2362, 9001 and 9002, B . thuringiensis H-14 and B . thuringiensis neoleonensis were tested for sensitivity against 18 antibiotics . The results revealed that all the four strains of B . sphaericus are resistant to colistin, nalidixic acid, polymyxin B and streptomycin . However, B . thuringiensis H-14 was resistant to 9 antibiotics, viz . ampicillin, cephalexin, carbenicillin, co-trimoxazole, colistin, cloxacillin, penicillin, nitrofurantoin and polymyxin B whereas B . thuringiensis neoleonensis was found to be resistant to 8 antibiotics . These results may help in isolation of potential and resistant mosquito pathogenic bacteria. Pneumoftiziologia, 1992 Oct-Dec, 41(4), 199 - 205 {The association of tuberculosis with HIV/AIDS infection in children in Romania}; Mihailescu P et al.; The great number of AIDS cases in children in Romania, together with the high annual risk of Tb infection, created the premises for the occurrence of a relatively great number of disease cases through HIV infection/AIDS + tuberculosis, particularly in the age-group "0-5 years" . Serum positive HIV children were considered as AIDS cases when tuberculosis was also associated . Twelve cases in which the infections were concomitant, transmitted through injections, constituted an exception to the point . The 12 children serum positive for HIV showed a primary musculo-cutaneous complex on their thighs, at the very place of injections . A proportion of 50% of them showed a favourable evolution under anti-Tb treatment . Most children developed primary tuberculosis aerogenically acquired, associated with AIDS . A proportion of 59.5% of them evoluted towards severe disseminated forms (milliaria, meningitis), with many deaths, and 37.8% only showed a favorable evolution under anti-tuberculosis treatment . HIV infection in children took place predominantly between 1987-1989 . Tuberculosis was associated 1-2 years later, when the switching from bacillary infection into active tuberculosis was facilitated by the progressive immunodepression which is specific for AIDS . The tuberculin test with 2 IU-PPD was positive in less advanced AIDS cases but faded in children in the final stage of the syndrome or in those with severe forms of tuberculosis . Tuberculosis finding out in children with HIV infection/AIDS is however possible; therefore, skin test reaction is compulsory in all children in this category . In children with a tuberculosis cured through specific treatment in their histories, the association of HIV infection reaching AIDS stage can lead to a Tb relapse.(ABSTRACT TRUNCATED AT 250 WORDS) Proteins . 1992 Oct;14(2):324. Purification and crystallization of insecticidal delta-endotoxin CryIIIB2 from Bacillus thuringiensis; Cody V et al.; CryIIIB2, an insecticidal protein from Bacillus thuringiensis has been crystallized from 0.6 M NaBr and HEPES buffer at pH 7.0 and X-ray diffraction data collected on a native crystal to 2.4 A . The insecticidal protein was obtained from a Bacillus thuringiensis (Bt) strain EG7231 . Crystals of the endotoxin are orthorhombic, space group C2221, with unit cell dimensions of a = 122.44, b = 131.81, and c = 105.37 A . A unit cell contains one molecule of the 67,000 Da endotoxin per asymmetric unit. Infect Agents Dis, 1992 Oct, 1(5), 245 - 53 Identification of uncultured microorganisms: expanding the spectrum of characterized microbial pathogens; Relman DA et al.; The combination of enzymatic nucleic acid amplification techniques with 16S rRNA-based molecular phylogeny has brought about a new approach to the identification of microbial pathogens that can not be cultivated in the laboratory . The applications of this experimental approach to bacillary angiomatosis and to Whipple's disease have revealed the presence of two previously uncharacterized organisms . These results suggest the existence of a far greater microbial diversity among human pathogens than has been so far appreciated with culture-dependent methods . PCR-based studies of aquatic environmental microbial communities have already reached similar conclusions . As a result, new and provocative questions are raised concerning the association of amplified 16S rRNA sequences with diseased tissue . The answers must await the results of further investigations and the expansion of sequence data bases. FEMS Microbiol Lett, 1992 Sep 15, 75(2-3), 143 - 8 Mode of replication, size and distribution of naturally occurring plasmids in Bacillus thuringiensis; Baum JA et al.; Cloned replication origin regions, derived from both small (4.9-7.5 MDa) and large (43-60 MDa) plasmids of Bacillus thuringiensis subspecies kurstaki strains HD73 and HD263 were used as hybridization probes in a Southern-blot analysis to assess both the size and horizontal distribution of native plasmid replicon groups among different subspecies of B . thuringiensis . In general, resident plasmids hybridizing to the replication origin regions derived from strains HD263 and HD73 were more commonly found in kurstaki strains than in non-kurstaki strains, suggesting a non-random distribution of plasmid incompatibility groups . Replication origin regions derived from the large HD263 plasmids (43-60 MDa) hybridized almost exclusively with large plasmids (greater than 30 MDa) of widely varying sizes . In contrast, replication origin regions derived from small plasmids hybridized exclusively with small plasmids (less than 10 MDa) showing little size variation . These results are consistent with previous observations concerning the relationship between plasmid size, mode of replication, and structural stability. J Biol Chem, 1992 Sep 25, 267(27), 19388 - 95 Molecular cloning and sequencing of the gene for mycocerosic acid synthase, a novel fatty acid elongating multifunctional enzyme, from Mycobacterium tuberculosis var . bovis Bacillus Calmette-Guerin; Mathur M et al.; Mycocerosyl lipids are found uniquely in the cell walls of pathogenic mycobacteria . Mycocerosic acid synthase (MAS) is a multifunctional protein which catalyzes elongation of n-fatty acyl-CoA with methylamalonyl-CoA as the elongating agent (Rainwater, D . L., and Kolattukudy, P . E . (1985) J . Biol . Chem . 260, 616-623) . To understand how the various domains that catalyze the reactions involved in chain elongation are organized, mas gene from Mycobacterium tuberculosis bovis BCG was cloned . A lambda gt11 library of AluI partially digested genomic DNA from the organism was screened with an oligonucleotide probe designed from the N-terminal amino acid sequence of purified MAS . Using terminal segments of inserts from positive clones as the probe, the library was rescreened and the process was repeated . Sequencing of four overlapping clones revealed a contiguous sequence of 9699 base pair(s) (bp) of mycobacterial genome containing a 6330-bp open reading frame that could code for a protein of 2100 amino acids with a molecular mass of 225,437 daltons . The authenticity of the open reading frame as that of MAS was verified by correspondence of the amino acid sequences deduced from the gene with the directly determined amino acid sequences of the N terminus and three different internal peptide fragments . By comparing the MAS amino acid sequence with the sequences in the active site regions of known fatty acid synthases and polyketide synthases the functional domains in MAS were identified . This analysis showed that the domains were organized in the following order: beta-ketoacyl synthase, acyl transferase, dehydratase-enoyl reductase, beta-ketoreductase, acyl carrier protein; no thioesterase-like domain could be found . These results establish MAS as the first case of an elongating multifunctional enzyme composed of two identical subunits that resemble the vertebrate fatty acid synthase in size, subunit structure, and linear organization of functional domains . Southern and Western blot analyses showed absence of mas gene and encoded proteins in Mycobacterium smegmatis and Escherichia coli . This result is consistent with the report that mycocerosic acid is present only in pathogenic mycobacteria. J Biol Chem, 1992 Sep 25, 267(27), 19278 - 90 Nucleotide sequence and deduced functions of a set of cotranscribed genes of Streptomyces coelicolor A3(2) including the polyketide synthase for the antibiotic actinorhodin; Fernandez-Moreno MA et al.; A 5.3-kb region of the Streptomyces coelicolor actinorhodin gene cluster, including the genes for polyketide biosynthesis, was sequenced . Six identified open reading frames (ORF1-6) were related to genetically characterized mutations of classes actI, VII, IV, and VB by complementation analysis . ORF1-6 run divergently from the adjacent actIII gene, which encodes the polyketide synthase (PKS) ketoreductase, and appear to form an operon . The deduced gene products of ORF1-3 are similar to fatty acid synthases (FAS) of different organisms and PKS genes from other polyketide producers . The predicted ORF5 gene product is similar to type II beta-lactamases of Bacillus cereus and Bacteroides fragilis . The ORF6 product does not resemble other known proteins . Combining the genetical, biochemical, and similarity data, the potential activities of the products of the six genes can be postulated as: 1) condensing enzyme/acyl transferase (ORF1 + ORF2); 2) acyl carrier protein (ORF3); 3) putative cyclase/dehydrase (ORF4); 4) dehydrase (ORF5); and 5) "dimerase" (ORF6) . The data show that the actinorhodin PKS consists of discrete monofunctional components, like that of the Escherichia coli (Type II) FAS, rather than the multifunctional polypeptides for the macrolide PKSs and vertebrate FASs (Type I). Biochim Biophys Acta, 1992 Sep 23, 1159(2), 185 - 92 Effects of pH on conformational properties related to the toxicity of Bacillus thuringiensis delta-endotoxin; Venugopal MG et al.; The delta-endotoxin of Bacillus thuringiensis subspecies kurstaki is an intracellular crystalline proteinaceous inclusion which, upon ingestion, is toxic to lepidopteran insects . Upon dissolution at pH > 9 it yields a protein subunit called protoxin . Under appropriate conditions, protoxin is hydrolyzed to a toxin molecule, which is responsible for killing the insect . It is known that this toxic activity decreases considerably above pH 10 . In this study, circular dichroism spectroscopy has been used to examine the secondary structures of the protoxin and toxin molecules at different pH values to determine if there are detectable conformational changes associated with their pH-dependent functional properties . At pH 10, where toxic activity is approximately maximal, both the protoxin and toxin molecules were found to assume a conformation that is on an average approx . 26% alpha-helix and approx . 45% beta-structure . As the pH was increased above 10, where the insecticidal activity decreases, the magnitude of the CD spectrum at 222 nm decreased for protoxin and the calculated alpha-helix contents of both protoxin and toxin molecules decreased . The net secondary structure did not change significantly at pH values below 10 . Significant conformational differences are observed between the secondary structure of the protoxin and toxin molecules at different pH values . The pH-dependent changes in secondary structure of the protoxin and toxin can be correlated with the effects of pH on the insecticidal activity of these proteins. Biochemistry, 1992 Sep 22, 31(37), 8740 - 6 Catalytic center of cyclodextrin glycosyltransferase derived from X-ray structure analysis combined with site-directed mutagenesis; Klein C et al.; An X-ray structure analysis of a crystal of mutant Asp229----Ala of cyclodextrin glycosyltransferase from Bacillus circulans (Ec 2.4.1.19) that had been shortly exposed to beta-cyclodextrin showed density corresponding to a maltose bound at the catalytic center . The crystal structure was refined to an R-factor of 18.7% at 2.5-A resolution . The catalytic center is defined by homology with the structurally known alpha-amylases and by the observation that mutants Asp229----Ala and Asp328----Ala are almost inactive . By model building, the density-defined maltose was extended to a full beta-cyclodextrin, which then indicated the general locations of seven subsites for glucosyl units . The catalytically competent residues Asp229, Glu257, and Asp328 are at the reducing end of the density-defined maltose . In the unligated wild-type structure, Glu257 and Asp328 form a 2.6-A hydrogen bond between their carboxylates in an arrangement that resembles those of the catalytically competent carboxylates in acid proteases . Presumably, the first catalytic step is an attack of the proton between Glu257 and Asp328 on the oxygen of the glycosidic bond. Biochemistry, 1992 Sep 22, 31(37), 8978 - 83 Substrate stereospecificity of phosphatidylinositol-specific phospholipase C from Bacillus cereus examined using the resolved enantiomers of synthetic myo-inositol 1-(4-nitrophenyl phosphate); Leigh AJ et al.; The substrate stereospecificity of phosphatidylinositol-specific phospholipase C from Bacillus cereus is examined using the resolved optical isomers of synthetic myo-inositol 1-(4-nitrophenyl phosphate), a chromogenic substrate for the phospholipase . The synthetic route employs mild acid-labile protecting groups and separation of the substituted myo-inositol enantiomers as the (-)-camphanyl ester diastereomers . Measurements of the initial rates of cleavage of the D and L enantiomers of the nitrophenyl substrate by phosphatidylinositol-specific phospholipase C from B . cereus show that this enzyme is essentially stereospecific for the D enantiomer . Under identical conditions, the rate of cleavage of the L isomer is less than 0.2% of that observed for the D isomer . The same is observed for the highly homologous enzyme from Bacillus thuringiensis . There is no measurable inhibition by the L enantiomer of the B . cereus enzyme acting on the D enantiomer, even when the molar ratio of L:D is 5, indicating that binding of the L enantiomer to the phospholipase is negligible . Thus, the enzyme active site is exquisitely sensitive to the stereochemistry of the myo-inositol group of the substrate. Gene, 1992 Sep 21, 119(1), 143 - 4 Sequence of the subtilisin-encoding gene from an antarctic psychrotroph Bacillus TA41; Davail S et al.; The nucleotide sequence of the subtilisin-encoding gene from the antarctic psychrotroph, Bacillus TA41, was determined . The primary structure of the subtilisin precursor corresponds to a preproenzyme of 419 amino acids . Asp144, His181 and Ser359 are the proposed catalytic residues of the protease active site. Biochem Biophys Res Commun, 1992 Sep 16, 187(2), 641 - 7 Genomic amplification and expression of delta-endotoxin fragment of Bacillus thuringiensis; Roy P; delta-Endotoxin gene of Bacillus thuringiensis HD-1 var kurstaki codes for the insecticidal crystal protein (ICP) specific for lepidopteran insects . Since the N-terminal half of the toxin is sufficient both for insect specificity and toxicity, the coding sequence of this part of the gene CryIA(b) was amplified by PCR and cloned in pUC19 . As there was no expression of immunologically detectable delta-endotoxin in this clone in E . coli, the amplified ICP gene was transferred to an expression vector pGEx2T . Restriction mapping and immunoblotting confirmed the presence and expression of the CryIA(b) gene . This insert should be suitable for expression in plant system if it is mobilized into a plant binary vector. Biochem J, 1992 Sep 15, 286 ( Pt 3), 857 - 62 Effect of side-chain amide thionation on turnover of beta-lactam substrates by beta-lactamases . Further evidence on the question of side-chain hydrogen-bonding in catalysis; Pratt RF et al.; Two side-chain-thionated beta-lactams, a penicillin and a cephalosporin, have been prepared and found to be not significantly poorer as substrates of typical serine (classes A and C) beta-lactamases than are their oxo analogues . This result is interpreted to mean that any hydrogen-bonding site on these enzymes for the beta-lactam side-chain amide carbonyl group must be flexible and is more likely to be a passive rather than active or essential feature of the active site . Previously, data from crystal structures and site-directed mutagenesis had suggested that the side chain of Asn-132 of class-A beta-lactamases, a component of the conserved SDN loop, forms a hydrogen bond with the side-chain carbonyl of the beta-lactam substrate and may provide significant transition-state stabilization during catalysis . The thionocephalosporin was also equally as good as its oxo analogue as a substrate of the class-B beta-lactamase II of Bacillus cereus and not significantly less effective as an inhibitor of the Streptomyces R61 DD-peptidase; a tight hydrogen-bond donor site for the beta-lactam side-chain amide is apparently not present in these enzymes either. FEMS Microbiol Lett, 1992 Sep 15, 75(2-3), 187 - 92 Occurrence of cold-labile NAD-specific glutamate dehydrogenase in Bacillus species; Jahns T; A nicotinamide adenine dinucleotide-specific glutamate dehydrogenase (NAD-GluDH; EC 1.4.1.3) inactivated by incubation at low temperatures was detected in several species of the genus Bacillus, including strains of B . cereus, B . laterosporus, B . lentus, B . panthotenicus, B . pasteurii, B . sphaericus, B . stearothermophilus, B . subtilis and B . thuringiensis . Incubation of cell-free extracts of these strains at 0 degrees C resulted in an 80-100% inactivation of NAD-GluDH activity within 120 min . The addition of 20% glycerol protected the enzyme from this inactivation in the cold . Strains of B . fastidiosus, B . licheniformis, B . macerans, B . megaterium and B . pumilus were found to lack NAD-GluDH activity. Appl Environ Microbiol, 1992 Sep, 58(9), 2820 - 6 Immunological demonstration of a unique 3,4-dihydroxyphenylacetate 2,3-dioxygenase in soil Arthrobacter strains; Olson PE et al.; Many bacteria biosynthesize 3,4-dihydroxyphenylacetate 2,3-dioxygenases for growth on aromatic acids, but gram-negative organisms have been most extensively studied . A gram-positive strain containing 2,3-dioxygenase activity was identified as Arthrobacter strain Mn-1 . The 2,3-dioxygenase from strain Mn-1 was purified to homogeneity by fast protein liquid chromatography with a Mono Q anion-exchange column . Rabbit polyclonal antidioxygenase antibodies were prepared . Ouchterlony double-diffusion and Western blotting (immunoblotting) protocols were used to probe the distribution of the Mn-1 dioxygenase antigen in soil bacteria . Fourteen 2,3-dioxygenase-containing Bacillus and Pseudomonas strains did not contain immunologically cross-reactive proteins . Six of eight Arthrobacter strains contained 2,3-dioxygenase activity, and all of them produced cross-reactive proteins . The data presented here suggest that a unique type of dioxygenase is geographically widespread but is taxonomically confined to Arthrobacter soil bacteria. JAMA, 1992 Sep 9, 268(10), 1280 - 6 Hospital outbreak of multidrug-resistant Mycobacterium tuberculosis infections . Factors in transmission to staff and HIV-infected patients; Beck-Sague C et al.; OBJECTIVE--To describe transmission of multidrug-resistant (MDR) Mycobacterium tuberculosis infection among patients and health care workers (HCWs) in a ward and clinic for human immunodeficiency virus (HIV)-infected patients in a hospital, four studies were conducted . METHODS--Case patients and control patients were persons who had been treated in the HIV ward or clinic, whose clinical course was consistent with tuberculosis and who had at least one positive culture for M tuberculosis between January 1, 1988, and January 31, 1990, resistant to at least isoniazid and rifampin (case patients), or whose isolates were susceptible to all drugs tested (control patients) . In the first study, case patients and control patients were compared to identify risk factors for MDR tuberculosis . In the second study, inpatient and outpatient days of MDR tuberculosis case patients were compared to determine whether acid-fast bacillus (AFB) smear-positivity or aerosolized pentamidine use was associated with higher numbers of subsequent MDR tuberculosis cases among exposed patients . In the third study, restriction fragment length polymorphism analysis was performed on available MDR and sensitive M tuberculosis isolates . In the fourth study, skin test conversion rates among HCWs in the HIV ward and clinic were compared with those of HCWs in another ward, and the strength of the associations between skin test conversions among HCWs on the HIV ward and the number of person-days that AFB smear-positive case patients and control patients were on this ward was estimated . RESULTS--Case patients were more likely than control patients to have been exposed on the HIV ward or clinic to an AFB smear-positive case patient (P less than .001) . Inpatient and outpatient days of MDR tuberculosis case patients were associated with more subsequent cases of MDR tuberculosis if exposing case patients were smear-positive or if they received aerosolized pentamidine (P less than or equal to .01) . Of 13 MDR isolates, all had one of two restriction fragment length polymorphism patterns; 10 sensitive isolates had restriction fragment length polymorphism patterns that were different from each other . The HCW skin test conversion rate was higher on the HIV ward and clinic than on the comparison ward (P less than .01) . The risk of occupational acquisition of infection increased in direct proportion to the number of person-days that AFB smear-positive case patients were on the HIV ward (r = .75; P = .005), but did not increase in proportion to the number of person-days that AFB smear-positive control patients were there (r = -.36; P = NS) . After isolation measures for AFB smear-positive tuberculosis patients were improved, MDR tuberculosis cases decreased to seven of 214 tuberculosis patients . CONCLUSIONS--Nosocomial transmission of MDR M tuberculosis infection to patients and HCWs occurred on the HIV ward and clinic . Infectiousness of MDR tuberculosis case patients was associated with AFB sputum-smear positivity . Case patients with MDR tuberculosis created a greater risk of skin test conversion for HCWs on the HIV ward than drug-susceptible control patients. Biochemistry, 1992 Sep 8, 31(35), 8307 - 14 Construction of a stable dimer of Bacillus stearothermophilus lactate dehydrogenase; Jackson RM et al.; A molecular graphics analysis of the features which prevent cytosolic malate dehydrogenase dimers from forming tetramers was evaluated by its success in predicting the synthesis of a version of the LDH framework which is a stable dimer . Surface residues responsible for malate dehydrogenases being dimers were revealed by superimposing the structures of two dimers of pig cytosolic malate dehydrogenase on one homologous tetramer of L-lactate dehydrogenase from Bacillus stearothermophilus . Four regions were identified as composing the P-axis dimer-dimer interface . Two regions of the dimer were surface loops that collided when built as a tetramer: a large loop (residues 203-207, KNOBI) and a small loop (residues 264-269, KNOBII), and these were candidates to explain the dimeric character of malate dehydrogenase . The analysis was tested by constructing a synthetic B . stearothermophilus lactate dehydrogenase (KNOBI) containing the large malate dehydrogenase loop (residues 203-207 being AYIKLQAKE, and extra four amino acids) . The new construct was thermotolerant (90 degrees C) and enzymically active with kcat and KM (pyruvate) values similar to those of the wild-type enzyme . However, whereas the allosteric activator fructose 1,6-bisphosphate decreased KM 100 times for wild type, it had no influence on KNOBI . The molecular volumes of 1-120 microM concentrations of the construct were measured by time-resolved decay of tryptophan fluorescence anisotropy and by gel filtration . Both methods showed the molecular weight of wild type increased from dimer to tetramer with Kd about 20 microM dimer . KNOBI remained a dimer under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS) J Leukoc Biol, 1992 Sep, 52(3), 303 - 6 Macrophage inflammatory proteins MIP-1 and MIP-2 are involved in T cell-mediated neutrophil recruitment; Appelberg R; Mice intraperitoneally (i.p.) infected with Mycobacterium bovis bacille Calmette-Guerin (BCG) respond to an i.p . challenge with mycobacterial antigen with an acute and extensive accumulation of neutrophils . This influx was not mimicked by the inoculation of recombinant interferon-gamma (IFN-gamma) or tumor necrosis factor alpha (TNF-alpha) . The antigen-induced recruitment of neutrophils was not affected by coinoculation of anti-IFN-gamma antibodies, was enhanced by anti-TNF-alpha antisera, and was significantly reduced by antisera against macrophage inflammatory proteins MIP-1 and MIP-2 . The latter two sera had no additive effects . We hypothesize that mycobacteria-specific T cells are triggered by antigen to secrete MIP-1 and MIP-2, which directly mediate, at least partly, the influx of neutrophils that takes place in this model. Am Rev Respir Dis, 1992 Sep, 146(3), 752 - 6 Factors associated with tuberculin reactivity among the foreign-born in Montreal; Menzies R et al.; Because tuberculosis among the foreign-born is of increasing importance in North America, it has recently been recommended that newly arriving immigrants be tuberculin tested and preventive therapy given to all those with significant reactions . The factors affecting the prevalence of tuberculin reactions were assessed in a community-based tuberculin survey among foreign-born schoolchildren and young adults . Of 1,198 foreign-born who were tuberculin tested, 32.4% had significant tuberculin reactions . False-positive tuberculin reactions due to sensitivity to purified protein derivative (PPD)-B (for Mycobacterium avium) were uncommon and those due to BCG vaccination of importance only among immigrants from countries with low tuberculosis rates . Tuberculin reactions of 10+ mm were associated with tuberculosis rates in the country of origin (p less than or equal to 0.001), age when immigrated (p less than or equal to 0.001), bacillus Calmette-Guerin (BCG) vaccination (p less than or equal to 0.01), and residence in poorer neighborhoods in Montreal (p less than or equal to 0.001), but not with number of years resident in Canada . The booster phenomenon, seen in 16% of those undergoing two-step testing, was most strongly associated with prior BCG vaccination (p less than or equal to 0.001) and also with tuberculosis rates in the country of origin (p less than or equal to 0.08), age of immigration (p less than or equal to 0.01), and number of years resident in Canada (p less than or equal to 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) Chest, 1992 Sep, 102(3), 972 - 4 Polypoid endobronchial lesions . A manifestation of bacillary angiomatosis; Slater LN et al.; Polypoid endobronchial lesions occurred in a patient with acquired immunodeficiency syndrome (AIDS) with recent fever, skin lesions, lymphadenopathy, lung infiltrates, and pleural effusions . His condition improved with antimicrobials and vincristine . After therapy ceased, skin lesions recurred and gastroesophageal mucosal lesions developed . Bacillary angiomatosis was identified during retrospective analysis of skin and endobronchial biopsy specimens. Clin Exp Immunol, 1992 Sep, 89(3), 402 - 6 The pattern of mycobacterial antigen recognition in sera from Mantoux-negative individuals is essentially unaffected by bacille Calmette-Guérin (BCG) vaccination in either south India or London; Das S et al.; Paired sera were obtained before and 8 weeks after routine BCG vaccination from 20 PPD-S Mantoux-negative individuals who were living adjacent to the Chingleput BCG vaccine trial area in Tamil Nadu and from seven Mantoux-negative school-children in London, UK . Most subjects became Mantoux-positive after vaccination . In ELISA tests against soluble extracts of BCG or Mycobacterium tuberculosis H37Rv or against PPD-S, pre-vaccination antibody titres of South Indian subjects were about twice those of British subjects but there was no increase in titre of antibodies after vaccination of either population . Western blotting showed that even before vaccination, and even in British subjects, antibodies were present that recognized numerous antigenic components in extracts of BCG and M . tuberculosis . There was no consistent difference between band patterns with South Indian and British subjects and any effect of vaccination on the patterns was minimal. J Urol, 1992 Sep, 148(3), 900 - 5 Bacillus Calmette-Guerin abrogates in vitro invasion and motility of human bladder tumor cells via fibronectin interaction; Garden RJ et al.; Intravesical bacillus Calmette-Guerin (BCG) has been shown to be an effective treatment for superficial transitional cell carcinoma of the bladder (TCC) . The mechanisms by which BCG limits tumor cell activity have thus far been unclear . We investigated the interaction between BCG and invasive human TCC cell line EJ in an in vitro invasion assay . We observed that BCG inhibited the invasion of EJ cells through an artificial basement membrane . In terms of the steps involved in tumor cell invasion, i.e . attachment, proteolysis, and motility, BCG was found to limit tumor cell motility . Attachment and proliferation of tumor cells were not affected by BCG . The effects of BCG on tumor cell migration were mediated by fibronectin (FN), a basement membrane glycoprotein component . Abrogation of BCG-FN-tumor cell interactions with anti-FN antibodies eliminated the ability of BCG to block tumor cell invasion . Fibronectin appears to link BCG and tumor cells via independent FN binding receptors to separate domains of the FN molecule . The molecular mechanism by which BCG may limit tumor cell motility may be its ability to protect against the formation of specific FN sequences as a result of protease cathepsin B digestion . A 31 kD and 27 kD FN band were absent from purified or tumor cell associated cathepsin B digestion when incubated in the presence of BCG, but present in the absence of BCG . Furthermore when purified from SDS polyacrylamide gel electrophoresis, the fragments were shown to have motility stimulating activity for the invasive EJ cells . These findings suggest that BCG functions as a potent inhibitor of tumor cell invasion . We conclude that BCG-fibronectin-tumor cell interactions may have a direct influence on the invasive mechanisms, such as motility, of tumor cells. J Urol, 1992 Sep, 148(3), 797 - 801 Management of stage T1 superficial bladder cancer with intravesical bacillus Calmette-Guerin therapy; Cookson MS et al.; We reviewed our results with 86 patients who had a pretreatment history of a stage T1 tumor . All patients were treated with transurethral resection of all visible tumor followed by intravesical bacillus Calmette-Guerin (BCG) and many patients received additional maintenance therapy . Local recurrences were treated with repeat transurethral resection followed by additional BCG . Median followup was 59 months, with a range of 9 to 149 months . Overall, 78 of 86 patients (91%) were free of tumor recurrence with BCG therapy . This result includes 69% of the patients who responded to the initial transurethral resection and intravesical BCG, and 22% who ceased having tumors after additional treatments for local recurrences . Only 7% of the patients had progression to stage T2 tumors after BCG therapy . Grade of the stage T1 tumor, concurrent carcinoma in situ and tumor multiplicity before BCG did not predict tumor recurrence or progression . Of patients with recurrences after BCG therapy, those with stage T1 tumors had a higher rate of progression compared to those with stage Ta tumors but the difference was not statistically significant (p = 0.086) . These data clearly support the efficacy of transurethral resection plus intravesical BCG immunotherapy in the treatment of stage T1 tumors as well as in the prevention of disease progression. Biochemistry, 1992 Sep 1, 31(34), 7787 - 95 Evidence for temperature-dependent conformational changes in the L-lactate dehydrogenase from Bacillus stearothermophilus; Kotik M et al.; L-Lactate dehydrogenase from Bacillus stearothermophilus (BSLDH) has been shown to change its conformation in a temperature-dependent manner in the temperature range between 25 and 70 degrees C . To provide a more detailed understanding of this reversible structural reorganization of the tetrameric form of BSLDH, we have determined in the presence of 5 mM fructose, 1,6-bisphosphate (FBP) the effect of temperature on far-UV and near-UV circular dichroism (CD), Nile red-binding to the enzyme surface, NADH binding, fluorescence polarization of fluorescamine-labeled protein, and hydrogen-deuterium exchange . In addition, we have analyzed the temperature dependence of the dimer-tetramer equilibrium of this protein by steady-state enzyme kinetics in the absence of FBP . The results obtained from these measurements at various temperatures can be summarized as follows . No changes in the secondary-structure distribution are detectable from far-UV CD measurements . On the other hand, near-UV CD data reveal that changes in the arrangements of aromatic side chains do occur . With increasing temperature, the asymmetry of the environment around aromatic residues decreases with a small change at 45 degrees C and a more pronounced change at 65 degrees C . Nile red-binding data suggest that the BSLDH surface hydrophobicity changes with temperature . It appears that decreasing the surface hydrophobicity may be a strategy to increase the protein stability of the active enzyme . We have noted significant alterations in the thermodynamic binding parameters of NADH above 45 degrees C, indicating a conformational change in the active site at 45 degrees C . The hydrodynamic volume of BSLDH is also temperature dependent.(ABSTRACT TRUNCATED AT 250 WORDS) Bioorg Khim, 1992 Sep, 18(9), 1186 - 9 {Site-specific endonuclease BstM6I from Bacillus stearothermophilus M6}; Shapovalova NI et al.; A site-specific endonuclease activity was found in extract of Bacillus stearothermophilus M6 isolated from molasses . The functionally pure enzyme designated as BstM6I was obtained by consecutive chromatographies on blue sepharose, hydroxyapatite, and heparin-sepharose . The endonuclease recognizes the nucleotide sequence CC decreases WGG in double-stranded DNA and cleaves it as indicated by the arrow to give one-nucleotide 5'-protruding ends . Consequently, the site-specific endonuclease BstM6I is an isoshizomer of BstNI. Trop Med Parasitol, 1992 Sep, 43(3), 191 - 4 Detection of Bartonella bacilliformis in cultures, blood, and formalin preserved skin biopsies by use of the polymerase chain reaction; Maass M et al.; A polymerase chain reaction (PCR) is described for the detection of Bartonella bacilliformis, the etiologic agent of bartonellosis, which cannot be identified biochemically . Amplification of a genomic 231 bp Bartonella DNA sequence permitted specific identification of 12 Bartonella isolates from Peruvian bartonellosis patients as well as detection of Bartonella DNA in blood samples and formaldehyde preserved skin biopsies . Specificity of amplification products was confirmed by restriction fragment analysis . No positive results were obtained with Brucella abortus, phylogenetically closely related to B . bacilliformis, and several other bacterial, fungal, and protozoal species . PCR appears as a promising technique for specific identification of B . bacilliformis in cultures and in clinical materials with further applications in taxonomic studies and in the investigation of Bartonella-like isolates obtained outside South America. Chem Pharm Bull (Tokyo), 1992 Sep, 40(9), 2478 - 82 Changes in enzymatic and membrane-adsorbing activities of sphingomyelinase from Bacillus cereus by modification with a polyethylene glycol derivative; Matsuyama H et al.; Sphingomyelinase (SMPLC) from Bacillus cereus was modified with a polyethylene glycol (PEG) derivative, methoxypolyethylene glycol-succinimidyl succinate (ss-PEG) . The molecular weight of the ss-PEG-modified SMPLC was calculated to be approx . 150 kDa by gel-filtration whereas that of the native enzyme, was 25 kDa . By this modification, the enzyme increased its thermostability and retained its hydrolytic activity toward 2-(N-hexadecanoylamino)-4-nitrophenylphosphocholine (HNP) and sphingomyelin (SM) in the mixed micelles with the surfactants such as Triton X-100 and sodium deoxycholate (SDC) . However, the activity toward liposomal SM was significantly decreased, and all the enzyme activities toward bovine erythrocytes, including membraneous SM-hydrolyzing and hemolytic activities as well as the enzyme adsorption onto the erythrocyte membranes, were completely lost. Br J Urol, 1992 Sep, 70(3), 271 - 5 Outcome in carcinoma in situ of bladder treated with intravesical bacille Calmette-Guérin; Harland SJ et al.; Fifty-three patients with carcinoma in situ of the bladder were treated with Evans strain BCG given intravesically . Complete remission was achieved after either one or two 6-weekly courses in 53% of patients . The median duration of remission was 32 months . Treatment-related bladder symptoms were minor during the first course, more severe during the second . There was no relation between severity of symptoms and likelihood of response . With a median follow-up of 32 months, disease progression has occurred in 10% of complete responders, whereas failure to respond on either cystoscopic, histological or cytological grounds was associated with a 48% progression rate . Although intravesical BCG produces impressive responses in carcinoma in situ of the bladder, managed conservatively the condition remains a dangerous one. Hinyokika Kiyo, 1992 Sep, 38(9), 1063 - 5 {Multiorgan failure following intravesical bacillus Calmette-Guerin administration: a case report}; Baba Y et al.; We describe a case of multiorgan failure after intravesical bacillus Carmette-Guern (BCG) immunotherapy for bladder cancer . A 58-year-old man with superficial transitional cell carcinoma of the bladder was initially treated by transurethral resection and intravenous chemotherapy, and then administered 11 sessions of BCG intravesically . He was administered BCG intravesically after cystoscopic examination . The next day he complained of nausea and malaise . He became hypotensive . The symptom progressed with multiorgan failure, disseminated intravascular coagulation and respiratory failure . The patient gradually improved with administration of antibiotics and corticosteroid, and hemodialysis, without antituberculous antibiotics . Intravesical instillation of BCG should not be carried out immediately after cystoscopic examination. J Clin Microbiol, 1992 Sep, 30(9), 2502 - 3 Bilophila wadsworthia bacteremia in two patients with hepatic abscesses; Kasten MJ et al.; Bilophila wadsworthia, a recently described anaerobic gram-negative bacillus, was isolated from blood cultures of two patients with liver abscesses . These isolates exhibited the expected phenotypic characteristics, and identification was confirmed by cell wall fatty acid analysis by gas-liquid chromatography . This is the first documentation of bacteremia due to B . wadsworthia. Biosci Biotechnol Biochem, 1992 Sep, 56(9), 1429 - 33 Effects of Bacillus thuringiensis var . israelensis 20-kDa protein on production of the Bti 130-kDa crystal protein in Escherichia coli; Yoshisue H et al.; A 20-kDa protein of