Mikrobiol Zh, 1989 Sep-Oct, 51(5), 26 - 8 {The effect of mineral salts on the growth and development of Bacillus thuringiensis strain H14 266/2-1}; Osadchaia AI et al.; Potassium phosphates in the concentration of 1.0 g/l are the most favourable for the growth and development of the culture of Bacillus thuringiensis H14 266/2-1 . The data obtained may be used when optimizing the medium for the submerged cultivation of a bactoculicide producer. Tubercle, 1989 Sep, 70(3), 179 - 86 Tuberculosis in British Columbia among immigrants from five Asian countries, 1982-85; Wang JS et al.; The influence of immigration from six selected Asian countries--Japan, Korea, Philippines, India, China and Hong Kong--on the incidence of tuberculosis in British Columbia has been examined . During the period 1982-1985 the average annual incidence of bacillary tuberculosis in these immigrants was more than six times as great as the overall British Columbia rate and contributed a quarter of the cases of active bacillary tuberculosis in this province whereas the immigrants from these countries represented only 3.7% of the total population . The clinical patterns of active tuberculosis by birthplace were analysed . A high proportion of cases of lymphadenitis was seen among all immigrants from Asia, particularly those from the Philippines . Both primary and secondary drug resistance was substantially higher than in Canadian-born patients . The frequency of primary drug resistance was higher among patients aged less than 40 years than those aged 40 or more. Scanning Microsc, 1989 Sep, 3(3), 843 - 53; discussion 853-4 Bacillus-shaped deposits composed of hexahedrally based crystals in human dental calculus; Kodaka T et al.; In human supra- and subgingival calculus, bacillus-shaped deposits showing various rocky-pile forms composed of hexahedrally based crystals were observed by scanning electron microscopy . The crystal size measured approximately 0.1-1.5 microns . The electron probe microanalysis always detected calcium, phosphorous, and magnesium . Their molar ratios resembled those of magnesium-containing whitlockite and moreover the crystals also gave the electron diffraction pattern of whitlockite . The bacillus-shaped deposits happened to coexist with the intracellular calcifying microorganisms, furthermore, oral microorganisms partially replaced by the hexahedrally based crystals were found . The crystal deposits were never seen in the surface layers of calculus exposed to the oral cavity, but occurred in the innermost layers and intra-spaces of supragingival and ledge-type subgingival calculus and in the outer layers of deep subgingival calculus. Mol Biol (Mosk), 1989 Sep-Oct, 23(5), 1455 - 68 {Localization of energy domains in Bacillus intermedius 7P ribonuclease}; Grishina IB et al.; Two independently melting regions (energetic domains) were localized in Bacillus intermedius 7P ribonuclease by methods of circular dichroism and high resolution X-ray analysis: the lov-temperature melting domain, containing C-terminal region of the molecule with five strands in antiparallel beta-structure and the high-temperature melting alpha-helical domain in the N-terminal region . The contact between these domains is stabilized mainly by ionic interaction Asp-22 - Lys+-48 . At pH 2.4 and 30.5 0 C, when the low-temperature domain melts, half of the beta-structure content in binase is destroyed though the alpha-helical structure content is conserved . It has been shown that in pH interval 2.4-4.8 at 15 0 C no changes in secondary structure and local surrounding of aromatic amino acid residues could be identified . Thus, the changes in ionic interactions in the binase molecule due to protonation of Asp side chain groups does not effect the secondary or tertiary structure, though it changes the energetical state of the binase molecule, revealing a change of number and size of energetic domains. Biokhimiia, 1989 Sep, 54(9), 1457 - 66 {Detection of a sodium pump in the terminal segment of the bacterial respiratory chain}; Verkhovskaia ML et al.; An alkalo- and halotolerant aerobic microorganism has been isolated which, according to microbiological data and the ribosomal 5S-RNA sequence, is a Bacillus similar, but not identical, to B . licheniformis and B . subtilis . The microorganism termed as Bacillus FTU proved to be resistant to the protonophorous uncoupler CCCP . The fast growth of Bacillus FTU in the presence of CCCP was shown to require high Na+ concentrations in the medium . A procedure has been developed to exhaust endogenous respiratory substrates in Bacillus FTU cells so that fast oxygen consumption by the cells was observed only upon addition of an exogenous respiratory substrate . The exhausted cells were found to oxidize ascorbate in the presence of TMPD in a cyanide-sensitive fashion . Ascorbate oxidation was coupled to the uphill Na+ extrusion stimulated by CCCP and a penetrating weak base, diethylamine (DEA), as well as by valinomycin with or without DEA . The operation of the Bacillus FTU terminal oxidase resulted in the generation of delta psi which, in a Na+ medium, was slightly decreased by CCCP and strongly by CCCP + DEA . In a K+ medium CCCP discharged delta psi even without DEA . Ascorbate oxidation was competent in ATP synthesis which was resistant to CCCP in the Na+ medium and sensitive to CCCP in the K+ medium . CCCP + DEA were inhibitory in both media . The data obtained indicate that there is a Na+-motive terminal oxidase in Bacillus FTU . It is suggested that delta microNa formed by the oxidase can be utilized by an Na+-driven ATP-synthase. Mikrobiol Zh, 1989 Sep-Oct, 51(5), 20 - 5 {Intensification of the growth and development of Bacillus thuringiensis H14 266/2-1 by optimizing the nutrient medium}; Osadchaia AI et al.; The mathematical method of experimental design was used to develop a new enzymic medium for cultivation of Bacillus thuringiensis H14 266/2-1, a bactoculicide producer . The optimized medium based on corn flour enzyme lysate as a carbon source and fodder yeast enzyme lysate as a source of nitrogen amine made it possible to increase twice the titre biomass yield for 24 h cultivation as compared to the initial medium . The above medium does not yield to the initial production medium in the insecticide activity. J Bacteriol, 1989 Sep, 171(9), 5141 - 7 Evidence for two different types of insecticidal P2 toxins with dual specificity in Bacillus thuringiensis subspecies; Nicholls CN et al.; Analysis of polypeptides in the crystalline delta-endotoxins from different Bacillus thuringiensis strains revealed two antigenically similar forms of the P2 protein which differed in molecular mass, peptide profile, and amino acid sequence . Purified preparations of the two forms displayed the characteristic dual toxicity of the P2 protein towards members of the orders Lepidoptera and Diptera in vivo but differed markedly in potency for the insects tested . Both species of the P2 protoxin, solubilized and activated by sequential proteolysis with insect gut extract and alpha-chymotrypsin, retained activity in vivo and in vitro, despite the removal of 144 residues from the N terminus . For the low-molecular-mass form, the dual insecticidal activity was reproducible in the in vitro assays. Appl Environ Microbiol, 1989 Sep, 55(9), 2428 - 30 Biosynthesis of 130-kilodalton mosquito larvicide in the cyanobacterium Agmenellum quadruplicatum PR-6; Angsuthanasombat C et al.; The 130-kilodalton mosquito larvicidal gene, cloned from Bacillus thuringiensis var . israelensis, was introduced into the cyanobacterium Agmenellum quadruplicatum PR-6 by plasmid transformation . Transformed cells synthesized 130-kilodalton delta-endotoxin protein and showed mosquito larvicidal activity . Results demonstrate a potential use of a cyanobacterium for biological control of mosquitoes. Mol Gen Mikrobiol Virusol, 1989 Sep, (9), 7 - 11 {Cloning the alpha-amylase gene of Bacillus amyloliquefaciens in cyanobacteria cells}; Elanskaia IV et al.; The recombinant plasmids of pIAH4amy series were constructed containing the alpha-amylase gene of Bacillus amyloliquefaciens A50 with its own promoter and leading sequence within an integrative vector plasmid pIAH4 (CmR) for cyanobacterium Anacystis nidulans R2 . At Anacystis nidulans transformation the hybrid plasmids integrate into cyanobacterium chromosome with high efficiency and all CmR transformants produce alpha-amylase . Expression of bacillar alpha-amylase gene in cyanobacterium cells is independent of the cloned gene orientation in the vector plasmid . Secretion of alpha-amylase into the cyanobacterial periplasm has been demonstrated. J Appl Bacteriol, 1989 Sep, 67(3), 275 - 82 A comparative study of enzyme variation in Bacillus cereus and Bacillus thuringiensis; Zahner V et al.; Thirty-two strains of Bacillus spp . were examined in a multilocus enzyme study by agarose gel electrophoresis . The organisms were Bacillus thuringiensis (21 strains, B . cereus (8), including two of var . mycoides, and B . megaterium (3) . Strains having similar enzyme variants were grouped into zymovars . A total of 10 of 11 enzyme loci studied were polymorphic and 27 zymovars were distinguished among the 32 strains . The results were subjected to numerical analysis, phenetic affinities and genetic distances between the strains were calculated . The numerical analysis was unable to differentiate between B . thuringiensis and B . cereus . Our results indicated that based on this multilocus enzyme study these zymovars should be considered as belonging to the same species . A mycoides variant of B . cereus was the most distinctive strain studied and clearly belonged to a separate species, B . mycoides . The technique also allowed for identification of contamination and mislabelling of strains. Nippon Seikeigeka Gakkai Zasshi, 1989 Sep, 63(9), 1117 - 23 {Effect of activated macrophages on cultured fibroblasts of the spinal posterior longitudinal ligament of the rabbit}; Miyata R; The present study was performed to elucidate the ossification of the posterior longitudinal ligament (OPLL) of the cervical spine . The relationship between OPLL and active oxygen was investigated using cultured fibroblasts derived from the spinal posterior longitudinal ligament of rabbit . Alveolar macrophages obtained from the same rabbit inoculated with Bacillus Calmette-Guerin (BCG) was added in the tissue culture system . The macrophages display a greatly enhanced power to generate O2- when stimulated by polystyrene latex spherules, 10-fold higher than treated with NaF . The cultured fibroblasts were directly connected with activated alveolar macrophages, then morphological changes and outgrowths were studied by phase contrast microscope . A significant effect was observed after in vitro exposure to 6 x 10(6) activated macrophages; the cells became greatly differentiated within 30 min, then proliferated and grew into multilayer after 6 days of culture . It is difficult to prove that the activated macrophages are a primary cause of OPLL, but the evidence presented here suggests that the active oxygen and other products derived from macrophages may give osteogenic potentiality to the cells in the spinal posterior longitudinal ligament. J Invertebr Pathol, 1989 Sep, 54(2), 200 - 7 Studies on the cellular defense reactions of the Madeira cockroach, Leucophaea maderae: nodule formation in response to injected bacteria; Rahmet-Alla M et al.; Nodules were formed in the Madeira cockroach, Leucophaea maderae, in response to injections of low doses (3 x 10(4) bacteria/insect) of three strains of Bacillus cereus and Escherichia coli K12 D31 . The most pathogenic strain of bacteria used, B . cereus B1, produced the greatest cellular response, while the least pathogenic, E . coli K12 D31, injected at the same dose, caused little nodule formation . Similarly, nodules were generally found to be larger following injection of pathogenic bacteria such as B . cereus B1 than to the weak pathogen, E . coli K12 D31 . There was, however, no difference in the extent of nodule formation with the four bacterial strains/species if they were heat killed prior to injection . Histologically, the nodules formed in response to all bacterial species employed were similar, with a central necrotic core enclosing cell debris and occasional bacteria, and an outer, thin sheath of plasmatocyte-like hemocytes . Possible reasons for the enhanced cellular reactivity observed in L . maderae to pathogenic bacteria are discussed. Trop Med Parasitol, 1989 Sep, 40(3), 251 - 7 Leprosy and tuberculosis vaccine design; Kaufmann SH; Tuberculosis and leprosy are bacillary infectious diseases which cause severe global health problems with approximately 50 to 60 million people suffering from tuberculosis and 10 to 15 million from leprosy . In the developing countries the currently available vaccine, Bacille Calmette-Guerin (BCG) was found to be less effective than originally thought . This disappointment, as well as recent achievements in biotechnology, has led several researchers to embark on novel avenues towards a rational vaccine design . This strategy stems from the idea that protective antigens exist which can be identified by immunological methods, expressed as recombinant gene products, and administered in a way that induces a protective T cell response. Prikl Biokhim Mikrobiol, 1989 Sep-Oct, 25(5), 658 - 63 {Various characteristics of lipid metabolism of Bacillus brevis var . G.-B.}; Zarubina AP et al.; Some characteristic features of the lipid metabolism of Bacillus brevis var . G.-B . natural variants and Bacillus brevis mutant 101 were studied . The authors found that upon submerged cultivation gramicidine S-producing P+-variant and B . brevis mutant 101 synthesized higher amounts of tocopherols as compared to other colonial-morphological variants . The highest tocopherol content was observed in P+-variant, whose cells contained the highest amount of total lipids as compared to other gramicidine S-producers. FEBS Lett, 1989 Aug 28, 254(1-2), 43 - 6 Addition of a methyl group changes both the catalytic velocity and thermostability of the neutral protease from Bacillus stearothermophilus; Takagi M et al.; Specific activity was compared between wild-type (WT) neutral protease from Bacillus stearothermophilus and mutant protease (M1; Gly144 replaced by Ala144) with enhanced thermostability . When casein was used as a substrate, M1 showed 1.5-times higher specific activity than that of WT . In contrast, the specific activities of M1 for soluble reduced lysozyme and insulin B chain were lower than those of WT by 17.2 and 13.2%, respectively . After digestion of the insulin A chain by these enzymes, the peptide products were purified and the N-terminal amino acid sequences were determined . WT enzyme cleaved insulin A chain at three sites, whereas no digestion was observed with M1 . Using Z-Gly-Leu-NH2 as a substrate, the kinetic parameters were determined . The Km values are nearly equal for both enzymes, whereas the kcat of M1 (240 min-1) was much smaller compared to the WT (830 min-1) . The data indicate that the mutation (addition of a methyl group) exerts an effect by changing both the catalytic velocity and thermostability. J Biol Chem, 1989 Aug 25, 264(24), 14386 - 8 Purification, crystallization, and preliminary x-ray diffraction studies of the flavoenzyme mercuric ion reductase from Bacillus sp . strain RC607; Moore MJ et al.; The flavoenzyme mercuric ion reductase from Bacillus sp . strain RC607 was purified by dye-ligand affinity chromatography . The protein was crystallized from solutions of high ionic strength, and one of the two crystal forms obtained has proven suitable for x-ray diffraction studies . Preliminary analysis showed that these crystals belong to the tetragonal space group 1422 . The unit cell dimensions are a = b = 180.7 A; c = 127.9 A . The diffraction pattern extends to better than 3 A resolution . Crystal density measurements are consistent with one enzyme dimer of 2 x 69,000 Da comprising the asymmetric unit . Trypsin treatment of the native enzyme resulted in the removal of 157 amino acids at the N terminus . After purification, the remaining fragment (amino acids 158-631), which is still fully active in vitro, could be crystallized under the same conditions as native enzyme . Twinning problems, however, did not allow complete analysis of these crystals. Eur J Biochem, 1989 Aug 15, 183(3), 671 - 8 The Na+-motive terminal oxidase activity in an alkalo- and halo-tolerant Bacillus; Semeykina AL et al.; An alkalo- and halo-tolerant aerobic microorganism has been isolated which, according to microbiological analysis data and the ribosomal 5S RNA sequence, is a Bacillus similar, but not identical, to B . licheniformis and B . subtilis . The microorganism, called Bacillus FTU, proved to be resistant to the protonophorous uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) . The fast growth of Bacillus FTU in the presence of CCCP was shown to require a high Na+ concentration in the medium . A procedure was developed to exhaust endogenous respiratory substrates in Bacillus FTU cells so that fast oxygen consumption by the cells was observed only when an exogenous respiratory substrate was added . The exhausted cells were found to oxidize ascorbate in the presence of N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) in a cyanide-sensitive fashion . The ascorbate oxidation was coupled to the uphill Na+ extrusion which was stimulated by CCCP and a penetrating weak base, diethylamine, as well as by valinomycin with or without diethylamine . Operation of the Bacillus FTU terminal oxidase resulted in the generation of a delta psi which, in the Na+ medium, was slightly decreased by CCCP and strongly decreased by CCCP + diethylamine . In the K+ medium, CCCP discharged delta psi even without diethylamine . Ascorbate oxidation was competent in ATP synthesis which was resistant to CCCP in the Na+ medium and sensitive to CCCP in the K+ medium as if Na+- and H+-coupled oxidative phosphorylations were operative in the Na+ and K+ media, respectively . Inside-out subcellular vesicles of Bacillus FTU were found to be competent in the Na+ uptake supported by oxidation of ascorbate + TMPD or diaminodurene . CCCP or valinomycin + K+ increased the Na+ uptake very strongly . The process was completely inhibited by cyanide or monensin, the former, but not the latter, being inhibitory for respiration . The data obtained indicate that in Bacillus FTU there is not only H+-motive but also Na+-motive terminal oxidase activity. Gene, 1989 Aug 15, 80(2), 363 - 8 A new isoschizomer, BnaI, of the BamHI restriction endonuclease; Kim EL et al.; By assaying the yield of phage SPO1 we have identified a new restriction-modification activity in the Bacillus natto B3364 strain . A class II restriction endonuclease, BnaI, isolated from the crude extract of B3364 cells was shown to be a true isoschizomer of the BamHI endonuclease . The Mr, stability and optimal conditions required for DNA digestion were determined for BnaI . Although both enzymes show the same specificity, BnaI and BamHI differ from each other in all the properties specified above. Biochim Biophys Acta, 1989 Aug 14, 1008(3), 293 - 300 Interactions of Escherichia coli SO-187 tRNA(IVal) with Bacillus stearothermophilus valine-tRNA synthetase studied by 13C-NMR; Schweizer MP et al.; Uracil isotopically labelled with 13C at C4 and C5 has been incorporated into nucleic acids of the Escherichia coli uracil auxotroph, SO-187 . {4,5-13C}uracil-labeled tRNA(IVal) was isolated and purified . 13C longitudinal relaxation times measured at 67.8 MHz demonstrated that the C5 dipole caused a 20-50% increase in the C4 relaxation . Interactions of this tRNA with valine-tRNA synthetase (VTS) purified from Bacillus stearothermophilus were established by 13C-NMR . Specific spectral changes were seen at 4-thiouridine, ribothymidine and pseudouridine of the 'bend' in the three-dimensional structure, and particularly at the uridine-5-oxyacetic acid in the wobble position of the anticodon . Thus, the protein seems to be in contact along the entire tRNA molecule, including the anticodon loop. Biochemistry, 1989 Aug 8, 28(16), 6605 - 10 On the effects of replacing the carboxylate-binding arginine-171 by hydrophobic tyrosine or tryptophan residues in the L-lactate dehydrogenase from Bacillus stearothermophilus; Luyten MA et al.; For L-lactate dehydrogenases (LDH's), the interaction of the guanidinium group of their Arg 171 residue with the carboxylate group of an alpha-keto acid is of primary importance in orienting the substrate productively at the active site . LDH's such as that of Bacillus stearothermophilus (BSLDH) are of practical importance for the preparation of chiral 2-hydroxy acids used as synthons in asymmetric synthesis but would even be more valuable in this regard if their specificities were broader . With a view to tailoring the specificity of BSLDH toward carbonyl substrates that lack an alpha-carboxyl group such as ketones, site-directed mutagenesis has been applied to replace Arg 171 by the approximately isosteric, but hydrophobic, amino acids Tyr and Trp . The mutant enzymes exhibit remarkably good catalytic activities toward representative alpha-keto acids RCOCOOH, where R = Me, Et, n-Pr, n-Bu, and CH2OH, although for the mutant enzymes the kcat/KM's are lower by approximately 10(3)-10(4)-fold than those for native BSLDH . Surprisingly, the 171----Tyr/Trp enzymes are significantly more active than 171----Lys (Hart et al., 1987a), for which an interaction of a positively charged side chain with substrate COO- is retained . Preparative-scale 171----Trp catalyzed reduction of pyruvate gave optically pure L-lactate, showing that L stereospecificity of such LDH enzymes was unaffected by the loss of Arg 171 . The retention of L stereospecificity is attributed to secondary polar or hydrogen-bonding associations of Arg 109 and Thr246, respectively, with the substrate COO-function that are of sufficient magnitude to maintain "normal" substrate orientation.(ABSTRACT TRUNCATED AT 250 WORDS) Hosp Pharm, 1989 Sep, 24(9), 701 - 4 The extended-spectrum penicillins: microbiologic, utilization, and cost review in a community hospital; Stein GE et al.; The extended-spectrum penicillins have a similar spectrum of activity and clinical efficacy . The newer acylaminopenicillins (azlocillin, mezlocillin, piperacillin) offer several potential therapeutic advantages over the carboxypenicillins (carbenicillin, ticarcillin), but they are more expensive per gram . Literature reviews of these penicillins suggest that individual hospital susceptibility patterns and cost should guide selection of the most appropriate antibiotic for formulary addition . Consequently, the authors performed a microbiologic, utilization, and cost review of these antibiotics in a community hospital . From this hospital and literature review, the authors were able to make the following conclusions: (1) the newer acylaminopenicillins exhibited similar activity against common clinical pathogens and were microbiologically more active than the carboxypenicillins; (2) none of these antibiotics used alone appears adequate for empiric treatment of serious systemic gram-negative bacillary infections; (3) the extended-spectrum penicillins were prescribed in a similar fashion and usually in conjunction with an aminoglycoside; and (4) significant cost savings can be realized when less expensive drugs are used . The authors found only minor differences in microbiologic activity and clinical use among the newer extended-spectrum penicillins . Therefore, they recommend the least expensive drug for routine use in the hospital. Mol Gen Mikrobiol Virusol, 1989 Aug, (8), 27 - 34 {Structure and biological features of the temperate phage of Bacillus thuringiensis var . galleriae 69/9, sensitive to chloroform}; Koretskaia NG et al.; A new temperate bacteriophage designated Px1 has been isolated from the culture of Bacillus thuringiensis var . galleriae 69/6 producing enthobacterin . The bacteriophage belongs to morphological group B1 in accordance with the classification by D . Reanney and H . Ackerman . The bacteriophage head has an isometric multifaceted form with 40 nm diameter . The length of its noncontractile transversely lined tail is 130 nm . High sensitivity to chloroform is peculiar of the phage . The lytical specter of the phage Px1 has been studied . The phage is shown to be capable of efficient transduction of plasmids between the bacteria of Bacillus cereus group. Fundam Appl Toxicol, 1989 Aug, 13(2), 310 - 22 Characterization of the mammalian toxicity of the crystal polypeptides of Bacillus thuringiensis subsp . israelensis; Mayes ME et al.; Solubilized crystal polypeptide preparations of Bacillus thuringiensis subsp . israelensis (BTI) were fractionated by immunoaffinity chromatography using a bound monoclonal antibody formed against the 28K crystal polypeptide . The 28K polypeptide was confirmed to be hemolytic and to possess low mosquitocidal activity against Aedes aegypti larvae . By comparison, the 28K polypeptide was more potent than the solubilized BTI crystals in male Swiss Webster mice, as the LD50 values were (p less than 0.05) 0.77 and 2.33 mg protein/kg body wt, respectively . Acute administration of the 28K polypeptide (mg/kg, ip) produced severe hypothermia and bradycardia in the mouse . No evidence for cooperativity between the 28K and other crystal polypeptides was observed . Preliminary histological examination of the mouse hearts exposed to the 28K polypeptide did not reveal any specific lesion, suggesting that the deficient cardiac performance might be a secondary physiological response . Gross pathological examination of mice as well as Sprague-Dawley rats acutely treated with equivalent doses of solubilized BTI crystal preparations revealed focal to segmental reddened and edematous areas within the small intestine . Histopathology indicated that the major lesion was in the jejunum . Contrary to expectations from in vitro hemolysis assays, cytolysis of mouse red and white blood cells was not detectable after in vivo exposure to the BTI solubilized proteins . The present results indicate that the 28K polypeptide is the mammalian toxic component of BTI crystals. Mol Gen Genet, 1989 Aug, 218(2), 355 - 7 Cloning and characterization of a gene cluster from Bacillus stearothermophilus comprising infC, rpmI and rplT; Pon CL et al.; Using two synthetic deoxyribonucleotide probes encoding segments of the primary structure of initiation factor IF3 from Bacillus stearothermophilus, we identified and cloned a segment of DNA which carries the infC gene . As in Escherichia coli, the infC gene begins with the unusual initiation triplet AUU, and is followed by the structural genes for ribosomal proteins L35 and L20 (rpmI and rplT, respectively). Urology, 1989 Aug, 34(2), 93 - 5 Squamous cell carcinoma of bladder after successful intravesical therapy with Bacillus Calmette-Guérin; Brenner DW et al.; A case of invasive squamous cell carcinoma of the bladder following intravesical immunotherapy with Bacillus Calmette-Guerin in a patient with pre-existing squamous dysplasia of the bladder is presented. Am J Clin Pathol, 1989 Aug, 92(2), 236 - 40 Cat-scratch disease in a patient with AIDS; Pilon VA et al.; A case of cat-scratch disease (CSD) in a patient with acquired immunodeficiency syndrome (AIDS) is reported . The lymph node pathologic characteristics were altered from those usually seen with CSD, showing clusters of vacuolated macrophages admixed with pycnotic nuclear debris instead of the usual suppurative granulomas . Evidence for the diagnosis was provided by Warthin-Starry stain and electron microscopic demonstration of the presumed CSD bacillus . Empiric treatment with antibiotics brought about clinical improvement . This case demonstrates the altered lymph node pathologic characteristics with CSD that may occur in a patient with AIDS. Taiwan Yi Xue Hui Za Zhi, 1989 Aug, 88(8), 828 - 31 Lung abscess caused by Eikenella corrodens: report of a case; Hsu CY et al.; Eikenella corrodens is a slow-growing, facultatively anaerobic, gram-negative bacillus . It is part of the normal flora of the human upper respiratory tract . We report a 68-year-old man who developed lung abscesses caused by E . corrodens while on long-term corticosteroid therapy for polymyositis . It was isolated by percutaneous transthoracic aspiration of an abscess under real-time sonographic guidance . The organism has a very unusual antimicrobial susceptibility: sensitive to penicillin, ampicillin, cephapirin, chloramphenicol, minocycline and erythromycin but resistant to clindamycin, oxacillin and gentamicin . The patient was treated empirically with penicillin G and gentamicin . The latter was discontinued after the results of the bacterial culture and sensitivity test were available . He was discharged one month later with marked improvement . Our report clearly demonstrates that E . corrodens can be the sole pathogen of a respiratory tract infection. Drugs, 1989 Aug, 38(2), 301 - 12 Chemotherapy in the management of bladder tumours; Whitmore WF Jr et al.; For patients with superficial bladder tumours intravesical treatment or prophylaxis with thiotepa, doxorubicin, mitomycin C or Bacillus Calmette-Guerin has added a useful dimension to management, although the precise indications for each regimen remain to be better defined . For patients with metastatic bladder cancer cisplatin and methotrexate (amethopterin), and to a lesser extent vinblastine and doxorubicin, are active single agents . Combinations of cisplatin and doxorubicin (adriamycin), and cisplatin and methotrexate +/- vinblastine +/- doxorubicin appear to induce complete remission in 20 to 35% of cases and partial remission in an additional 20 to 40% of cases . In some patients, complete remission has persisted from 2 to more than 10 years . Few randomised Phase III studies have been carried out to determine the relative effectiveness of different drug combinations, but the results of Phase II trials have encouraged investigations of adjuvant and neoadjuvant programmes combining such regimens with radiation or surgery, or both, in patients with clinically localised muscle infiltrating tumours. Br J Exp Pathol, 1989 Aug, 70(4), 435 - 41 Changes in phospholipid fatty acid composition and triacylglycerol content in mouse tissues after infection with bacille Calmette-Guérin; Jackson SK et al.; Changes in the lipids of tissues from mice infected with bacille Calmette-Guerin (BCG) have been detected by gas-liquid chromatography . Infection with BCG resulted in (1) an increase in the polyunsaturated to saturated fatty acid ratio of phospholipids and (2) a decrease in the total triacylglycerol fatty acid content of spleen, liver and peritoneal macrophages . The alteration in fatty acid composition was significant in the phosphatidylethanolamine fraction of the phospholipids . The relation of these findings to an increased sensitivity to bacterial endotoxins is discussed. J Bacteriol, 1989 Aug, 171(8), 4178 - 88 Cloning and sequencing of the gene encoding a 125-kilodalton surface-layer protein from Bacillus sphaericus 2362 and of a related cryptic gene; Bowditch RD et al.; Using the vector pGEM-4-blue, a 4,251-base-pair DNA fragment containing the gene for the surface (S)-layer protein of Bacillus sphaericus 2362 was cloned into Escherichia coli . Determination of the nucleotide sequence indicated an open reading frame (ORF) coding for a protein of 1,176 amino acids with a molecular size of 125 kilodaltons (kDa) . A protein of this size which reacted with antibody to the 122-kDa S-layer protein of B . sphaericus was detected in cells of E . coli containing the recombinant plasmid . Analysis of the deduced amino acid sequence indicated a highly hydrophobic N-terminal region which had the characteristics of a leader peptide . The first amino acid of the N-terminal sequence of the 122-kDa S-layer protein followed the predicted cleavage site of the leader peptide in the 125-kDa protein . A sequence characteristic of promoters expressed during vegetative growth was found within a 177-base-pair region upstream from the ORF coding for the 125-kDa protein . This putative promoter may account for the expression of this gene during the vegetative growth of B . sphaericus and E . coli . The gene for the 125-kDa protein was followed by an inverted repeat characteristic of terminators . Downstream from this gene (11.2 kilobases) was an ORF coding for a putative 80-kDa protein having a high sequence similarity to the 125-kDa protein . Evidence was presented indicating that this gene is cryptic. J Ultrastruct Mol Struct Res, 1989 Aug, 102(2), 178 - 87 Three-dimensional structure of the surface protein layer (MW layer) of Bacillus brevis 47; Tsuboi A et al.; The three-dimensional (3D) structure of one surface protein layer from Bacillus brevis 47, the middle wall (MW) layer, has been reconstructed from tilted-view electron micrographs after correlation averaging to a resolution of 2 nm . The MW layer has p6 symmetry with a center-to-center spacing of 18.3 nm and a minimum thickness of 5.5 nm . The reconstruction reveals a distinct domain structure: the heavier domain of six monomers jointly forms a massive core centered at the sixfold symmetry axis, and lighter domains interconnect adjacent unit cells . In addition, the larger domains collectively form a pore by making contact with each other towards the inner surface, while the smaller domains establish a second connectivity towards the outer surface of the S layer . The MW layer of B . brevis resembles the S layer of Acetogenium kivui in various aspects: they have very similar lattice parameters and highly reminiscent 3D structures; the pores penetrate through the whole core and appear to determine the porosity of the S layers. J Biochem (Tokyo), 1989 Aug, 106(2), 209 - 15 Nucleotide sequence of the glutamine synthetase gene (glnA) and its upstream region from Bacillus cereus; Nakano Y et al.; We have determined the complete nucleotide sequence of a 2.4 kb chromosomal EcoT22I-NspV fragment, containing the Bacillus cereus glnA gene (structural gene of glutamine synthetase) . The deduced amino acid sequence indicates that the glutamine synthetase subunit consists of 444 amino acid residues (50,063 Da) . Comparisons are made with reported amino acid sequences of glutamine synthetases from other bacteria . Upstrem of glnA we found an open reading frame of 129 codons (ORF129) preceded by the consensus sequence for a typical promoter . Maxicell experiments showed two polypeptide bands, with molecular weights in good agreement with that of glutamine synthetase and that of ORF129, in addition to vector-coded protein . It is possible that the product of this open reading frame upstream of glnA has a regulatory role in glutamine synthetase expression. J Invest Dermatol, 1989 Aug, 93(2), 287 - 90 Dispase, a neutral protease from Bacillus polymyxa, is a powerful fibronectinase and type IV collagenase; Stenn KS et al.; Dispase, a neutral protease isolated from culture filtrates of Bacillus polymyxa, has proven to be a rapid, effective, but gentle agent for separating intact epidermis from the dermis and intact epithelial sheets in culture from the substratum . In both cases it effects separation by cleaving the basement membrane zone region while preserving the viability of the epithelial cells . Because it is not known what or where in the basement membrane zone Dispase cleaves, we set up studies to define its substrate specificity . Using purified basement membrane components and sodium dodecyl sulfate-polyacrylamide gel electrophoresis we show that Dispase cleaves fibronectin and type IV collagen, but not laminin, type V collagen, serum albumin, or transferrin . The action of Dispase on collagen appears to be selective for type IV collagen in that several stable degradation products are formed, whereas the enzyme degrades type I collagen only minimally . In newborn human skin, as seen by electron microscopy, Dispase removes the lamina densa, rich in type IV collagen, but preserves the anchoring fibrils (structures known to contain type VII collagen) and the epidermal cells . Because its action is so selective, it suggests that Dispase can serve as a powerful tool for dissecting epithelial-mesenchymal interactions. Zhonghua Jie He He Hu Xi Za Zhi, 1989 Aug, 12(4), 225 - 7, 255 {Slide culture and rapid sensitivity test for tubercle bacillus}; Zhao H; We use the slide culture and direct sensitivity test for tubercle bacillus . The sputum specimens was taken from 479 cases . The positive rates of slide culture method and thick smear were 50.5% and 47.8% respectively . The positive rates from slide culture and modified Loewenstein-Jensen medium were 48.2% and 51.9% respectively . The results of sensitivity test of both methods in 40 cases were quite similar . The slide culture technique is a simple, cheap and very rapid method for sensitivity test, it is particularly useful for treatment of pulmonary tuberculosis patients. Can J Microbiol, 1989 Aug, 35(8), 760 - 3 L-alanine and inosine enhancement of glucose triggering in Bacillus megaterium spores; Bedard J et al.; Both rate and extent of germination of Bacillus megaterium 14581 (ATCC) spores are considerably augmented when L-alanine and inosine are added to the glucose commonly used as triggering agent for this strain . This enhancement does not arise from heterogeneity in germination requirements of the dormant spore, but is rather a consequence of the combined action of glucose and either or both of the added reagents on a sizeable fraction of spores unable to germinate in glucose alone . Nearly half of the spores that eventually germinate in the mixture of germinants used are either triggered by glucose or are sensitized by it to subsequent triggering by L-alanine and inosine in the first 10 s of imbibition . For a good number of these spores, then, triggering consists of a sequence of separable events. Kekkaku, 1989 Aug, 64(8), 499 - 509 {Analysis of interferon-gamma production in killed BCG-pretreated mice after stimulation with staphylococcal enterotoxin A}; Mizukoshi N et al.; Staphylococcal enterotoxin A (SEA), a T cell mitogen, was found to induce a high level of interferon-gamma (IFN-gamma) in mice which had been immunized with killed Bacillus Calmette-Guerin (BCG) in water-in-oil-in-water (W/O/W) emulsion . The phenomenon was analysed by in vivo and in vitro experiments, and the following results were obtained . 1 . The SEA-induced IFN was inactivated by treatment with 0.2M glycine-HCl (pH 2.0) but not by heating at 56 degrees C for 30 min . nor by treatment with anti-IFN alpha/beta antibodies, and the fact suggest that the IFN belonged to the gamma type . 2 . Treatment of the BCG-sensitized mice with silica and 2-chloroadenosine (2CA), specific lethal agents for macrophages, reduced the SEA-induced IFN production . 3 . The SEA-induced IFN production occurred in mice immunized with BCG either intravenously or intraperitoneally, although they showed weak or no footpad reaction to purified protein derivatives (PPD) . In contrast, mice sensitized subcutaneously with BCG showed strong foodpad reaction to PPD but not the SEA-induced IFN production . Thus, the mere presence of BCG-sensitized T cells does not appear to result in the SEA-induced IFN production . 4 . In vitro experiments, in which SEA-induced IFN production was determined in the culture of BCG-sensitized spleen cells, showed that principal IFN-producing cells were Lyt-1+ T cells . 5 . Deprivation of macrophages from BCG-sensitized spleen cell population by passing through Sephadex G-10 column reduced the SEA-induced IFN production in the culture . Addition of 2CA to the culture medium also reduced the SEA-induced IFN production by the BCG-sensitized spleen cells . 6 . The SEA-induced IFN production in the culture of the BCG-sensitized spleen cells was suppressed in the presence of lipoxygenase inhibitor, i.e., caffeic acid or nordihydroguaiaretic acid . 7 . The plastic adherent spleen cells (i.e . macrophages) from mice sensitized with BCG produced leukotriene C4 (LTC4) . The suppression of the SEA-induced IFN production of BCG-sensitized spleen cells in the presence of the lipoxygenase inhibitor was prevented by addition of synthetic LTC4 . These results suggest that LTC4 released from macrophages activated by BCG causes production of IFN-gamma by BCG-sensitized T cells responding to SEA. J Pak Med Assoc, 1989 Aug, 39(8), 199 - 201 Bacillus sphaericus as mosquito larvicide; Aziz A et al.; In Noorpur Shahan, a village in the outskirts of Islamabad, Bacillus sphaericus was tested to determine its efficacy against mosquito larvae . Since the creation of this new Islamabad district no mosquito control measure has been taken in the area and like so many other places in and around Islamabad, mosquito density is unusually high in this village . The efficacy of Bacillus sphaericus was studied upto seven weeks after its application and it gave good larval control. J Biochem (Tokyo), 1989 Aug, 106(2), 270 - 3 Role of outer coat in resistance of Bacillus megaterium spore; Nishihara T et al.; The outer coat fraction (OC-Fr) of Bacillus megaterium ATCC 12872 spore was isolated as a resistant residue after alkali extraction, sonic treatment, and pronase digestion of the spore coat preparation, and its backbone structure was determined by chemical analysis to be composed of galactosamine-6-phosphate (GalN-P) polymers with polypeptides and calcium . OC-Fr was not fully solubilized after ordinary acid hydrolysis . OC-Fr was insensitive to all hexosaminidases tested, and moreover, an isolated fragment, a pentamer of GalN-P, was also resistant to lysozyme and hexosaminidases even after N-acetylation, being sensitive to them to some extent after dephosphorylation . Molecular sieving experiments revealed that the outer coat limited the entry of compounds with a molecular weight of more than 2,000 . Exchange of the metal on the spore surface also influenced the heat resistance . Spores of OC-Fr-deficient mutants were less resistant but were still much more resistant than the vegetative cells . These results suggest that the outer coat protects the contents of the spore against chemical, physical and enzymatic treatments owing to the chemical structure itself, composed mainly of GalN-P polymers, and the molecular sieving effect. Br J Urol, 1989 Aug, 64(2), 143 - 6 Conservative treatment of diffuse carcinoma in situ of the bladder with repeated courses of intravesical therapy; Mukamel E et al.; We present a series of 13 patients with diffuse carcinoma in situ (CIS) of the bladder who failed an initial induction course of intravesical therapy with Mitomycin C, thiotepa, doxorubicin or Bacillus Calmette Guerin (BCG) . Cystectomy, although indicated, was, for various reasons, not performed after the first failure of intravesical therapy and all patients were subsequently treated topically with the same or different agents . Of the 7 patients treated with 2 induction courses, 6 showed a complete response during a follow-up period of 24 to 42 months . Although 1 patient initially responded completely, he developed invasive transitional cell carcinoma (TCC) Grade IV 30 months later . Among the 3 patients who underwent 3 induction courses, 2 had a complete response at 42 and 60 months of follow-up and 1 developed TCC Grade IV with muscle invasion 18 months later . Two of the 3 patients treated with 4 induction courses are free of disease at 48 and 57 months; the third developed low grade, low stage TCC . This experience suggests that the majority of patients with CIS who fail initial treatment usually respond to further treatment with the same or a different drug . The question as to whether a second course of intravesical therapy, subsequent to failure of the first course, should be given before cystectomy requires further investigation. FEBS Lett, 1989 Jul 17, 251(1-2), 183 - 6 Synthesis and secretion of bacterial alpha-amylase by the yeast Saccharomyces cerevisiae; Kovaleva IE et al.; Alpha-amylase from Bacillus amyloliquefaciens, synthesized in yeast Saccharomyces cerevisiae without substitution of the signal sequence, is efficiently secreted from yeast cells: 60-70% of the overall amount of the enzyme is found in the culture fluid . In contrast to many yeast secretory proteins, which accumulate in the periplasmic space and in the cell wall, intracellular alpha-amylase is localized mainly in the cytoplasm . Obviously, transfer across the cell wall is not a rate-limiting step in alpha-amylase export from the cell . The glycosylated forms of proteins are predominantly found both inside the cell and in the culture medium. FEMS Microbiol Lett, 1989 Jul 15, 51(1), 211 - 7 Transformation and expression of a cloned delta-endotoxin gene in Bacillus thuringiensis; Lereclus D et al.; A shuttle vector containing the replication region of a resident plasmid of B . thuringiensis, was used to determine the conditions allowing efficient transformation of B . thuringiensis by electroporation . Using this plasmid a delta-endotoxin gene was cloned and expressed both in Escherichia coli and B . thuringiensis . It was shown that this gene was poorly expressed in the wild type situation whereas after cloning in acrystalliferous strains of B . thuringiensis large amounts of crystal protein were obtained. Am J Ophthalmol, 1989 Jul 15, 108(1), 53 - 6 Microbial analysis of contact lens care systems contaminated with Acanthamoeba; Donzis PB et al.; We analyzed bacterial and fungal contamination within the contact lens care systems of ten patients who had Acanthamoeba detected within their care systems . Seven patients had Acanthamoeba keratitis, one had Pseudomonas keratitis, and the remaining two were asymptomatic . Gram-negative bacteria were found in all ten care systems, and Pseudomonas was found in six . Bacillus species, the only gram-positive bacteria isolated, were found in five systems . Fungi were isolated in six care systems . The use of homemade saline and the two-cup method of peroxide disinfection were associated with microbial contamination . Acanthamoeba organisms were found only in contact lens cases or solutions that also had bacterial and in many cases fungal contamination, suggesting that the presence of bacterial and fungal contamination within the contact lens care system may be an important element for the survival and growth of Acanthamoeba. J Biol Chem, 1989 Jul 15, 264(20), 11682 - 7 Mutations affecting the catalytic activity of Bacillus cereus 5/B/6 beta-lactamase II; Lim HM et al.; Random in vitro mutagenesis of a cloned Bacillus cereus 5/B/6 beta-lactamase II gene was used to select defective genes unable to confer ampicillin or cephalosporin C resistance to Escherichia coli . DNA sequencing of mutant genes identified histidine at position 28 as important to beta-lactamase II function . In addition, the isolation of six identical frameshift mutants established that the carboxyl-terminal end of beta-lactamase II is critical for enzyme function . Random mutagenesis also revealed that His88 (implicated previously as one of 4 residues acting as a zinc ligand) is crucial to enzymatic activity and that a glycine to glutamic acid substitution at position 148 produced a defective beta-lactamase . Oligonucleotide mutagenesis directed at Glu37 and Glu212 suggests that these residues are inconsequential to enzyme function but that histidine at position 28 may be involved in substrate binding or recognition. Biochem Biophys Res Commun, 1989 Jul 14, 162(1), 475 - 82 Biosynthesis of paf-acether . XIV . Paf-acether output in murine peritoneal macrophages is regulated by the level of acetylhydrolase; Palmantier R et al.; Paf-acether (paf) synthesis was previously shown to be impaired in 24 hr-adherent and Bacillus Calmette-Guerin-activated murine peritoneal macrophages as compared to resident macrophages . We report here that the induction of acetylhydrolase was responsible for the decreased paf output in 24 hr-adherent macrophages . The kinetic analysis of the enzymes derived from 2 hr-, 24 hr- and BCG-activated adherent macrophages and from plasma revealed that the Km for paf was similar whatever the source of the acetylhydrolase whereas the Vmax was five-fold increased in 24 hr-cultured macrophages . The acetylhydrolase activity was Ca2+-independent and was not inhibited by addition of alkyl-acyl (long chain)-glycero-phosphocholine suggesting that the enzyme was not a phospholipase A2. Brain Res, 1989 Jul 10, 491(2), 390 - 3 Demonstration of functional acetylcholinesterase on the soma of individual neurones of Aplysia by in vivo microspectrophotometry; Fossier P et al.; The presence of functional acetylcholinesterase is demonstrated in vivo on somatic membranes of single ganglionic neurones of Aplysia using concurrently microspectrophotometry and electrophysiology . The similarity of the effects of an irreversible blocker of acetylcholinesterase and of phospholipase C from Bacillus cereus suggests that acetylcholinesterase is anchored in the membrane via phosphatidylinositol. J Theor Biol, 1989 Jul 10, 139(1), 117 - 28 Phospholipid flip-out controls the cell cycle of Escherichia coli; Norris V; Phospholipids are the principal constituents of biological membranes . In Escherichia coli, phospholipids are involved in the metabolism of other envelope constituents such as lipoprotein, lipopolysaccharide, certain envelope proteins and peptidoglycan . They are also involved in the regulation of the cell cycle . DNAA, the key protein in the initiation of chromosome replication, is activated by acidic phospholipids only when these are in fluid bilayers, whilst interruptions of phospholipid synthesis inhibit both the initiation of chromosome replication and cell division . The transmembrane movement or flip-flop of phospholipids from one monolayer to the other requires the passage of the polar head group through the hydrophobic core of the bilayer . Hence, in many systems, flip-flop is a slow process with half-time of days . Flip-flop accompanies the formation of non-bilayer structure . Such structures form under certain conditions of packing density and composition and have been observed both in vitro and in vivo . In bacteria, flip-flop appears to be extremely rapid, with half-times as fast as 3 min being observed . However, such rapid flip-flop may not be characteristic of all phospholipids . The asymmetrical distribution of phosphatidylethanolamine in the plasma membrane of Bacillus megaterium has been attributed to the existence of two classes of this phospholipid . In E . coli, studies of the metabolic turnover of phosphatidylserine, phosphatidylglycerol and phosphatidic acid also reveal the existence of distinct classes of these phospholipids . In this article I propose that, in E . coli, a class of phospholipids does indeed escape the rapid flip-flop mechanism; this class probably includes a subpopulation of the acidic phospholipids . Therefore during the cell cycle these phospholipids accumulate in the inner monolayer of the cytoplasmic membrane and so cause an increase in its packing density; at a critical density, phospholipids "flip out" from the inner to the outer monolayer . This flip-out occurs once per cycle and initiates cell cycle events. J Mol Biol, 1989 Jul 5, 208(1), 183 - 94 Functional mapping of an entomocidal delta-endotoxin . Single amino acid changes produced by site-directed mutagenesis influence toxicity and specificity of the protein; Haider MZ et al.; Mutagenesis has been used to investigate the toxicity and specificity of a larvicidal protein from Bacillus thuringiensis aizawai IC1 that is toxic to both lepidoptera and diptera and differs by only three residues from a monospecific lepidopteran toxin from B . thuringiensis berliner . Site-directed mutagenesis was used to investigate the contribution of these residues to the dual specificity of the aizawai protein . The results suggest that changes in the identity of residues adjacent to Arg544 and Arg567 on the C-terminal side may convert a monospecific toxin into a dual specificity toxin by altering the protease sensitivity of the arginyl peptide bond . A series of deletion mutants was constructed and their protein products analysed for toxicity in vitro and in vivo and for their ability to perturb phospholipid bilayers . The results indicate a different functional role for various protein segments in the toxin's mode of action and suggest that two separate regions close to the C terminus of the active toxin are important in conferring dual specificity on the aizawai IC1 toxin . A model suggesting a basis for the activity of monospecific and dual-specificity B . thuringiensis toxins is presented, which postulates that association of sequences at the C terminus of the active toxin with regions near the N terminus may be responsible for determining toxin specificity. J Biol Chem, 1989 Jul 5, 264(19), 10987 - 95 Coding nucleotide, 5' regulatory, and deduced amino acid sequences of P-450BM-3, a single peptide cytochrome P-450:NADPH-P-450 reductase from Bacillus megaterium; Ruettinger RT et al.; Cytochrome P-450BM-3 (P-450BM-3) from Bacillus megaterium incorporates both a P-450 and an NADPH:P-450 reductase in proteolytically separable domains of a single, 119-kDa polypeptide and functions as a fatty acid monooxygenase independently of any other protein . A 5-kilobase DNA fragment which contains the gene encoding P-450BM-3 was sequenced . A single continuous open reading frame starting at nucleotide 1541 of the 5-kilobase fragment correctly predicted the previously determined NH2-terminal protein sequences of the trypsin-generated P-450 and reductase domains and, in toto, predicted a mature polypeptide of 1,048-amino acid residues with Mr = 117,641 . The trypsin site was found at arginine residue 471 . The P-450 domain is most similar (about 25%) to the fatty acid omega-hydroxylases of P-450 family IV, while the reductase domain exhibits some 33% sequence similarity with the NADPH:P-450 reductases of mammalian liver . Both the P-450 and reductase domains of P-450BM-3 define new gene families but contain highly conserved segments which display as much as 50% sequence similarity with P-450s and reductases of eukaryotic origin . The mRNA for P-450BM-3 was found by S1 mapping to be 3,339 +/- 10 nucleotides in length . In the accompanying paper, two regions in the 1.5 kilobases 5' to the P-450BM-3 coding region have been implicated in the regulation of P-450BM-3 gene expression. J Mol Biol, 1989 Jul 5, 208(1), 83 - 98 Structure of tyrosyl-tRNA synthetase refined at 2.3 A resolution . Interaction of the enzyme with the tyrosyl adenylate intermediate; Brick P et al.; The crystal structure of tyrosyl-tRNA synthetase (EC 6.1.1.1) from Bacillus stearothermophilus has been refined to a crystallographic R-factor of 22.6% at 2.3 A resolution using a restrained least-squares procedure . In the final model the root-mean-square deviation from ideality for bond distances is 0.018 A and for angle distances is 0.044 A . Each monomer consists of three domains: an alpha/beta domain (residues 1 to 220) containing a six-stranded beta-sheet, an alpha-helical domain (248 to 318) containing five helices, and a disordered C-terminal domain (319 to 418) for which the electron density is very weak and where it has not been possible to trace the polypeptide chain . Complexes of the enzyme with the catalytic intermediate tyrosyl adenylate and the inhibitor tyrosinyl adenylate have also been refined to R-factors of 23.9% at 2.8 A resolution and 21.0% at 2.7 A resolution, respectively . Formation of the complexes results in some crystal cracking, but there is no significant difference in the conformation of the polypeptide chain of the three structures described here . The relative orientation of the alpha/beta and alpha-helical domains is similar to that previously observed for the "A" subunit of a deletion mutant lacking the C-terminal domain . Differences between these structures are confined to surface loops that are involved in crystal packing . Tyrosyl adenylate and tyrosinyl adenylate bind in similar conformations within a deep cleft in the alpha/beta domain . The tyrosine moiety is in the equivalent position to that occupied by tyrosine in crystals of the truncated mutant and makes similar strong polar interactions with the enzyme . The alpha-phosphate group interacts with the main-chain nitrogen of Asp38 . The two hydroxyl groups of the ribose form strong interactions with the protein . The 2'-hydroxyl group interacts with the carboxylate of Asp194 and the main-chain nitrogen of Gly192 while the 3'-hydroxyl interacts with a tightly bound water molecule (Wat326) . The adenine moiety appears to make no significant polar interactions with the protein . The results of site-directed mutagenesis studies are examined in the light of these refined structures. J Biol Chem, 1989 Jul 5, 264(19), 10996 - 1003 Requirement for a 1-kilobase 5'-flanking sequence for barbiturate-inducible expression of the cytochrome P-450BM-3 gene in Bacillus megaterium; Wen LP et al.; In a previous publication (Wen, L.-P., and Fulco, A . J . (1987) J . Biol . Chem . 262, 6676-6682), we described the cloning of the gene encoding cytochrome P-450BM-3, a catalytically self-sufficient fatty acid monooxygenase induced by barbiturates in Bacillus megaterium . We have now subcloned a 1.6-kilobase segment of DNA from this cloned gene that includes the barbiturate-responsive regulatory region as well as 88 bases encoding the NH2-terminal portion of cytochrome P-450BM-3 . From this, we generated two series of 5' and 3' deletion derivatives and examined their effects on gene expression . When the 1.6-kilobase fragment or the 1.3-kilobase 5'----3' deletion derivative is inserted into Escherichia coli on a vector containing a promoterless chloramphenicol acetyltransferase (CAT) gene with the sequence encoding the NH2-terminal portion of P-450BM-3 placed immediately in front of the CAT gene, CAT activity is constitutive and unaffected by pentobarbital . On the other hand, the basal level of CAT is low in B . megaterium transformed by the same construct but is strongly inducible by pentobarbital . Furthermore, the multicopy plasmid containing this regulatory region causes a dramatic decrease in both the basal and pentobarbital-induced expression of chromosomally encoded P-450BM-3 in B . megaterium . This competition effect, unlike CAT expression, is independent of the orientation of the regulatory DNA segment in the plasmid . Removal of 0.3 kilobase or more from the 3' end of the 1.6-kilobase segment of DNA or 0.6 kilobase from the 5' end abolishes the competition effect and also eliminates basal and inducible CAT expression in B . megaterium . In transformed E . coli, constitutive CAT expression is maintained when as little as 0.3 kilobase of DNA from the 3' end of the 1.6-kilobase segment is inserted in the correct orientation in front of the CAT gene . The data are consistent with the hypothesis that the synthesis of cytochrome P-450BM-3 in B . megaterium is under positive control and requires gene interaction with at least one trans-acting factor, presumably a protein, to activate transcription from the P-450BM-3 promoter . The binding of this putative protein is mediated by at least two regulatory regions (R1 and R2) that span about 1 kilobase of the 5'-flanking region of the gene. Biophys Chem, 1989 Jul, 33(3), 257 - 64 Phosphorescence properties of Trp-84 and Trp-310 in glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus; Gabellieri E et al.; The phosphorescence spectra of Trp-84 and Trp-310 in glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus in an aqueous glass show distinct 0,0 vibrational bands with peaks at 406.5 and 410.5 nm . With the aid of external heavy-atom perturbation of iodide and the thermal quenching profile, it is concluded that although both chromophores are effectively buried, only one, viz., the 406.5 nm component, is embedded in a sufficiently rigid core of the protein to phosphoresce in fluid solutions at room temperature . From inspection of the crystallographic structure is it evident that only Trp-310 embedded in the beta-sheet of the catalytic domain may satisfy the requirements of a long triplet-state lifetime and slow migration of O2 to its site . This identification confirms previous analysis of the phosphorescence properties of the enzymes from yeast, pig and rabbit muscle. J Invertebr Pathol, 1989 Jul, 54(1), 63 - 70 Effect of exposure of Pieris brassicae larvae to 2,4,5-trichlorophenoxyacetic acid on the natural antibacterial activity of serum; Jones RD et al.; Larvae of the cabbage white butterfly, Pieris brassicae, were reared on a semisynthetic diet with or without 20 ppm of the herbicide 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and using three assays the sera were subsequently tested for natural antibacterial activity against Bacillus cereus, Escherichia coli K12, and Micrococcus luteus . These assays showed that exposure of larvae to 2,4,5-T lowered the antibacterial activity of the serum against E . coli and M . luteus compared with control animals . Spectrophotometric tests for the presence of a lysozyme-like principle in the serum also revealed similar trends with a significant loss of enzyme activity in 2,4,5-T-treated insects . Overall total serum protein levels of control and 2,4,5-T-treated insects were similar, suggesting a specific effect of the herbicide on certain serum components such as lysozyme . The possible mode of action of the herbicide on production of antibacterial factors is discussed. J Invertebr Pathol, 1989 Jul, 54(1), 57 - 62 Laboratory evaluation of three mosquito pathogenic strains of Bacillus sphaericus isolated in Egypt; Gharib AH et al.; Three strains of Bacillus sphaericus H-5a5b designated Ghar . 1 & 10, Ghar . 2 & 20, and Ghar . 3 & 30 were tested for growth, virulence, and larvicidal activity against Culex pipiens in the laboratory . Incubation temperature was positively correlated (r = 0.91) to the rate of bacterial growth (strain Ghar . 2 & 20) . All three strains retained their virulence through 25 successive transfers on nutrient agar . Acetone powder preparations showed high larvicidal activity against C . pipiens, although second instar larvae were more susceptible than fourth instars to all three strains . The most active strain was Ghar . 2 & 20 with LC50 values of 0.51 mg/liter (second instar) and 1.62 mg/liter (fourth instar) after 48 hr of exposure . Mortality rates in fourth instar larvae exposed to an acetone powder form of strain Ghar . 2 & 20 were significantly greater at higher than at lower temperatures. J Invertebr Pathol, 1989 Jul, 54(1), 16 - 22 Functional relationships between free amino acids in the hemolymph of fourth instar larvae of the mosquito Aedes aegypti (Diptera, Culicidae) as a basis for toxicological studies; Bounias M et al.; Characteristic correlations reflecting particular metabolic interactions between free amino acids have been pointed out and used as a sensitive test for the detection of biochemical intoxication symptoms in fourth instar larvae of the mosquito Aedes aegypti exposed for 0-8, 12, 24, and 36 hr to various doses of Bacillus thuringiensis delta-endotoxin ranging from 0.01 to 1 mg liter-1 . In a first pool, serine was negatively correlated with glycine while cystine and alanine were positively correlated with serine and proline . A second pool was characterized by strong positive correlations between leucine, valine, isoleucine, and threonine . These two groups were linked by a negative (hyperbolic) correlation between cystine and threonine . Preliminary data then gave evidence that the slopes of the linear regressions of Gly on Ser and Ala on Ser increased and those of Ile on Thr and Val on Leu decreased with increasing doses of B . thuringiensis israelensis delta-endotoxin . Functional relationships thus exhibited a high semiological value in metabolic stress studies. Arch Biochem Biophys, 1989 Jul, 272(1), 237 - 44 Formation of tyrosine O-sulfate by mitochondria and chloroplasts of Euglena; Saidha T et al.; Mitochondria that have been purified from cells of light-grown wild-type Euglena gracilis Klebs var . bacillaris Cori or dark-grown mutant W10BSmL and incubated with 35SO4(2-) and ATP accumulate a labeled compound in the surrounding medium . This compound is also labeled when mitochondria are incubated with {14C}tyrosine and nonradioactive sulfate under the same conditions . This compound shows exact coelectrophoresis with synthetic tyrosine O-sulfate at pH 2.0, 5.8, and 8.0, and yields sulfate and tyrosine on acid hydrolysis . Treatment with aryl sulfatase from Aerobacter aerogenes yields sulfate and tyrosine but no tyrosine methyl ester; no hydrolysis of tyrosine methyl ester to tyrosine is observed under identical conditions, ruling out methyl esterase activity in the aryl sulfatase preparation . Thus the compound is identified as tyrosine O-sulfate . No tyrosine O-sulfate is found outside purified developing chloroplasts of Euglena incubated with 35SO4(2-) and ATP, but both chloroplasts and mitochondria accumulate labeled tyrosine-O-sulfate externally when incubated with adenosine 3'-phosphate 5'-phospho{35S}-sulfate (PAP35S) . Since tyrosine does not need to be added, it must be provided from endogenous sources . Labeled tyrosine O-sulfate is found in the free pools of light-grown Euglena cells grown on 35SO4(2-) or in dark-grown cells incubated with 35SO4(2-) in light, but none is found in the medium after cell growth . No labeled tyrosine O-sulfate is found in Euglena proteins (including those in extracellular mucus) after growth or incubation of cells with 35SO4(2-) or after incubation of organelles with 35SO4(2-) and ATP or PAP35S, ruling out sulfation of the tyrosine in protein or incorporation of free-pool tyrosine O-sulfate into protein . The system forming tyrosine O-sulfate is membrane-bound and may be involved in transporting tyrosine out of the organelles. Taiwan Yi Xue Hui Za Zhi, 1989 Jul, 88(7), 669 - 72 Evaluation of humoral immunity on leprosy patients in Taiwan: a preliminary report; Wang CR et al.; Twenty-four tuberculoid (T)-type and 31 lepromatous (L)-type leprosy patients from Taiwan Provincial Lo-Sheng Leprosarium were enrolled in this study . Twenty-six age- and sex- matched normal subjects were also studied as a control group . The evaluation of their general and specific humoral immunity included B-cell subpopulations, 3 major classes of immunoglobulin (G, A and M) and antibodies in the IgG class against lepromin suspension and Bacillus Calmette-Guerin (BCG) sonicate . T-type patients showed a larger B-cell percentage than L-type patients (p less than 0.01) . In general, patients with leprosy, both T and L types, had higher serum immunoglobulin levels than the control group . T-type patients showed greater antibody levels than the control group (p less than 0.05 for anti-lepromin and p less than 0.0001 for anti-BCG) . L-type patients demonstrated a higher anti-BCG IgG level than the control group (p less than 0.0001) . The level of anti-BCG IgG was more frequently above the cutoff level than that of anti-lepromin IgG in leprosy patients (p less than 0.01 for T, p less than 0.005 for L) . In conclusion, humoral immunity is not impaired in leprosy patients . Discrepancies for T- and L-type patients among B-cell subpopulation, serum immunoglobulin levels and specific antibody levels reflect different aspects of cell-mediated immunity impairment . Though leprosy patients had elevated anti-BCG IgG levels, it is impossible to differentiate L- and T-type patients; specific antigens are needed for serodiagnosis of leprosy patients in Taiwan. J Infect Dis, 1989 Jul, 160(1), 104 - 15 Immunotherapy of localized, intermediate, and diffuse forms of American cutaneous leishmaniasis; Convit J et al.; The clinical efficacy of immunotherapy for localized American cutaneous leishmaniasis with a combination of heat-killed Leishmania mexicana amazonensis promastigotes and viable BCG (bacille Calmette Guerin) has been compared with meglumine antimoniate chemotherapy and with BCG alone in a controlled clinical study in 217 patients . The results in the first two groups were comparable, with greater than 90% clinical cures with an average time of 16-18 w required for healing . The cure rate was considerably lower (42%) and more prolonged in the group receiving BCG alone . Secondary effects were observed in less than 5% of the patients receiving combined immunotherapy or BCG alone . In contrast, 49% of the patients receiving chemotherapy showed side effects . High therapeutic efficacy was also observed using combined immunotherapy in patients with intermediate and diffuse cutaneous leishmaniasis who were previously unresponsive to chemotherapy . Cure or clinical improvement was seen in all 11 patients with intermediate forms of the disease, and marked clinical improvement was observed in 9 of 10 patients with diffuse disease . The results on the efficacy of the combined vaccine in immunotherapy for American cutaneous leishmaniasis provide a strong rationale for studying its effectiveness in prophylactic trials. Indian J Chest Dis Allied Sci, 1989 Jul-Sep, 31(3), 233 - 6 Immunoglobulin status of geriatric pulmonary tuberculosis patients of Himachal Pradesh; Arora VK et al.; Immunoglobulins (IgG, IgA & IgM) levels were estimated in 50 bacillary cases of geriatric pulmonary tuberculosis (PTB) and 25 healthy controls of comparable age group by single radial immunodiffusion technique . All Igs were raised in PTB cases, the rise being directly proportional to the radiological extent of disease . Exudative cases had more marked rise compared to patients with productive and fibrotic lesions . Those cases of PTB having associated COPD (28 cases), showed less marked increase in Igs and with treatment, Ig levels declined, the decline was slower in cases having associated COPD. Mol Biol (Mosk), 1989 Jul-Aug, 23(4), 1051 - 6 {Determination of the substrate specificity of Bpu101 restrictase with an unusual recognition segment}; Degtiarev SKh et al.; A new enzyme Bpu10I was isolated from Bacillus pumilus . This enzyme is not an isoschizomer of any known restriction endonucleases . The search of possible recognition sequences was carried out in sequences ABCNiDEF (i = 0.6) on substrate DNA lambda CI857, T7, pBR322 . The recognition sequence and cleavage sites of restriction endonuclease Bpu10I have been determined as CCTNAGC . GGANTCG Sci China B, 1989 Jul, 32(7), 830 - 6 Molecular cloning and expression of Bacillus thuringiensis subsp . galleriae insecticidal crystal protein genes in Escherichia coli; Chen Q et al.; The location of the toxin gene of B . thuringiensis subsp . galleriae (H5ab) on the Mr-130Md plasmid is determined by molecular cloning . Double digestion fragments (BamHI and SalI) and PstI restriction fragments as well, from the 130 Md plasmid, of B . thuringiensis subsp . galleriae, are ligated with the cloning vector pAT 153 respectively and transformed into E . coli strain HB 101 . Out of 208 transformants, three colonies (FG2, FG9, FG19) give positive hybridization reaction using the HD-1 delta-endotoxin gene as a probe . They are presumed to contain the delta-endotoxin gene of B . thuringiensis subsp . galleriae . Western blot assays indicate that Mr-130 kDal and 68 kDal, crystal proteins produced by clone FG2 react with anticrystal protein antibody . The protein extracts of clone FG2 are lethal to Ostrinia furnacalis (Guenee) . This is the first report with regard to the cloning and expression of the B . thuringiensis subsp . galleriae (H5ab) delta-endotoxin gene. Anal Biochem, 1989 Jul, 180(1), 99 - 104 A spectrophotometric procedure for measuring oxoglutarate and determining aminotransferase activities using nicotinamide adenine dinucleotide phosphate-linked glutamate dehydrogenase from algae; Ahmad I et al.; A new spectrophotometric procedure is described for determining glutamate-dependent activities of aspartate aminotransferase, alanine aminotransferase, and ornithine aminotransferase with NADPH-linked glutamate dehydrogenase (GDH) from nitrate-grown Stichococcus bacillaris . The algal NADPH-GDH is highly specific for oxoglutarate and can catalyze the reduction of this keto acid in the presence of high glutamate concentrations, and thus is suitable for the measurement of oxoglutarate produced in glutamate-dependent amino-transferase reactions . The alga produces large amounts of NADPH-GDH which can be adequately purified in a few simple steps . The purified enzyme can be stored at 4 degrees C for several weeks without any detectable loss of activity . The algal NADPH-GDH can also be used for the estimation of small amounts of oxoglutarate in aqueous extracts. Mol Gen Genet, 1989 Jul, 218(1), 177 - 81 Efficient transformation of Bacillus thuringiensis and B . cereus via electroporation: transformation of acrystalliferous strains with a cloned delta-endotoxin gene; Schurter W et al.; Electroporation was used as a method to transform intact cells of Bacillus thuringiensis and B . cereus . With our optimized method a range of plasmid vectors could be transformed into strains of B . thuringiensis at frequencies of up to 10(7) transformants/micrograms DNA . This high frequency allows cloning experiments to be done directly in B . thuringiensis . A bifunctional vector capable of replicating in Escherichia coli and in Bacillus spp . was constructed . The kurhd1 protoxin gene was cloned into this shuttle vector to produce plasmid pX193, then transformed into B . thuringiensis HD1 cryB and B . cereus 569K . The cloned protoxin gene was expressed in sporulating cultures of both strain HD1 cryB (pX193) and 569K (pXI93), producing crystal protein active in biotests against larvae of Heliothis virescens . This demonstrates the usefulness of the electroporation method for the introduction of cloned toxin genes, in either their native or modified form, into a variety of host strains. Biochem J, 1989 Jul 1, 261(1), 99 - 105 Proteolytic processing of a coleopteran-specific delta-endotoxin produced by Bacillus thuringiensis var . tenebrionis; Carroll J et al.; Insecticidal protein delta-endotoxin crystals harvested from sporulated cultures of Bacillus thuringiensis var . tenebrionis contain a major polypeptide of 67 kDa and minor polypeptides of 73, 72, 55 and 46 kDa . During sporulation, only the 73 kDa polypeptide could be detected at stage I . The 67 kDa polypeptide was first detected at stage II and increased in concentration throughout the later stages of sporulation and after crystal release, with a concomitant decrease in the 73 kDa polypeptide . This change could be blocked by the addition of proteinase inhibitors . Trypsin or insect-gut-extract treatment of the delta-endotoxin crystals after solubilization resulted in a cleavage product of 55 kDa with asparagine-159 of the deduced amino acid sequence of the toxin {Hofte, Seurinck, van Houtven & Vaeck (1987) Nucleic Acids Res . 15, 71-83; Sekar, Thompson, Maroney, Bookland & Adang (1987) Proc . Natl . Acad . Sci . U.S.A . 84, 7036-7040; McPherson, Perlak, Fuchs, Marrone, Lavrik & Fischhoff (1988) Biotechnology 6, 61-66} at the N-terminus . This polypeptide was found to be as toxic in vivo as native delta-endotoxin. Mikrobiologiia, 1989 Jul-Aug, 58(4), 553 - 6 {Substrate specificity of enzymes from Bacillus mesentericus}; Nesterova NG et al.; The proteolytic enzymes of the sporogenous Bacillus mesentericus strains 64 and 8 were tested for their ability to hydrolyse different protein substrates . The enzymes were isolated using affinity chromatography on bacillichine-silochrome, and eluted with 25% isopropanol in 0.05 M Tris-HCl buffer, pH 8.0-8.4, containing 0.01 M CaCl2 . Casein, hemoglobin, elastin, albumin and synthetic peptides, Z-L-Ala-Ala-Leu-pNa and Z-L-Ala-Gly-Leu-pNa, were used as substrates . The activity of esterase was assayed in terms of indophenyl acetate cleavage . The proteinases were compared with terrilytin, a commercial preparation . The proteinase of strain 64 was active in the hydrolysis of casein, hemoglobin and elastin; its specificity was close to that of terrilytin . The proteinase of strain 8 differed from them in a higher thrombolytic and fibrinolytic activity, and had a high esterase activity. Eur J Immunol, 1989 Jul, 19(7), 1303 - 10 Differential pattern of T cell recognition of the 65-kDa mycobacterial antigen following immunization with the whole protein or peptides; Brett SJ et al.; The 65-kDa stress protein from Mycobacterium bovis (Bacillus Calmette Guerin) elicited T cell proliferation and antibody responses in seven B10 congenic mouse strains with different H-2 haplotypes . To analyze T cell determinants on this antigen, seven peptides corresponding to six predicted T cell epitopes, and one defined B cell epitope were synthesized . Mice were either immunized with the whole antigen and the specificity of the response was ascertained in respect of the six peptides, or mice were immunized with seven of the peptides and tested for proliferative responses to the whole molecule . The results showed that three peptides carried epitopes to which mice responded following injection of the whole molecule and that immunization with two additional peptides could prime for in vitro stimulation with the native antigen . The latter result indicates the feasibility of generating T cell responses to "cryptic" epitopes on proteins by immunizing with peptides . The peptide-specific T cell responses were distinctly influenced by the H-2 haplotype of mouse strains . However, two peptides were recognized by several H-2-disparate mouse strains, and one peptide could be presented by both I-A and I-E molecules . Immunization with several peptides induced a cross-reactive T cell proliferative response to the homologous GroEL protein isolated from E . coli . The amount of cross-reactivity was influenced by the extent of sequence homology between mycobacterial and E . coli proteins and the major histocompatibility complex class II molecule used to present the peptide. Biochem Biophys Res Commun, 1989 Jun 30, 161(3), 1126 - 34 Expression of an NCA cDNA in NIH/3T3 cells yields a 110K glycoprotein, which is anchored into the membrane via glycosyl-phosphatidylinositol; Kolbinger F et al.; The NCA cDNA, which represents a gene belonging to the CEA family, was inserted into an SV40 early promoter-driven expression vector and used for transfection of mouse NIH/3T3 cells . A cell line, NIH/3T3/KNCA IG7, was selected which expressed a molecule with an apparent molecular weight of 110,000 . The mode of membrane attachment of this NCA, which we already proposed to be anchored via glycosyl-phosphatidylinositol, was investigated by treatment of NIH/3T3/KNCA IG7 cells with phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis . Two independent methods, flow cytometry and immunoprecipitation of {3H}-labelled surface glycoproteins, clearly demonstrated that the NCA molecule expressed by NIH/3T3/KNCA IG7 cells is indeed anchored into the membrane via glycosyl-phosphatidylinositol . Furthermore, these results support our previous biochemical data on NCA-50, by unequivocally showing that the NCA cDNA used for transfection encodes an NCA molecule related to NCA-50 and NCA-90. Biochem Biophys Res Commun, 1989 Jun 30, 161(3), 1273 - 9 Functions of the COOH-terminal region of cyclodextrin glucanotransferase of alkalophilic Bacillus sp . #1011: relation to catalyzing activity and pH stability; Kimura K et al.; Cyclodextrin glucanotransferase (beta-CGTase) of alkalophilic Bacillus sp . #1011 degrades starch to mainly beta-cyclodextrin (beta-CD) . This enzyme is considered to contain an extra-polypeptide in its COOH-terminal region in addition to its NH2-terminal domain which exhibits the starch-degrading activity . To analyze the functions of this extra-polypeptide in the beta-CGTase, two mutated enzymes, in which DNA regions encoding 10 or 13 amino acids from the COOH-terminus were deleted, were obtained . The mutated enzymes degraded starch to glucose, maltooligosaccharides and alpha-CD, in addition to beta-CD . Furthermore, the pH stability of the mutated enzymes in the alkaline pH range (pH 9-11) was reduced. J Immunol Methods, 1989 Jun 21, 120(2), 215 - 20 Development of a fluorescent immunodot assay for Bacillus cereus enterotoxin; Jackson SG; A fluorescent immunodot assay has been developed for rapid, specific detection of B . cereus enterotoxin . None of the other Bacillus species tested showed cross-reactivity in the assay with antiserum to purified B . cereus enterotoxin . The assay can detect greater than or equal to 50 ng of purified enterotoxin . Using this assay system, enterotoxin was found to be produced by 25 of 25 foodborne disease-related isolates and 22 of 25 isolates not related to foodborne disease (isolates from routine surveillance foods) . Because of the apparent widespread ability of isolates to produce enterotoxin the assay may have potential as a rapid identification procedure for B . cereus . The substrate gel system described may have wider application in other immunoassay systems using a membrane solid phase. J Mol Biol, 1989 Jun 20, 207(4), 805 - 21 Crystal structure of unliganded phosphofructokinase from Escherichia coli; Rypniewski WR et al.; In an attempt to characterize the mechanism of co-operativity in the allosteric enzyme phosphofructokinase from Escherichia coli, crystals were grown in the absence of activating ligands . The crystal structure was determined to a resolution of 2.4 A by the method of molecular replacement, using the known structure of the liganded active state as a starting model, and has been refined to a crystallographic R-factor of 0.168 for all data . Although the crystallization solution would be expected to contain the enzyme in its inactive conformation, with a low affinity for the co-operative substrate fructose 6-phosphate, the structure in these crystals does not show the change in quaternary structure seen in the inactive form of the Bacillus stearothermophilus enzyme (previously determined at low resolution), nor does it show any substantial change in the fructose 6-phosphate site from the structure seen in the liganded form . Compared to the liganded form, there are considerable changes around the allosteric effector site, including the disordering of the last 19 residues of the chain . It seems likely that the observed conformation corresponds an active unliganded form, in which the absence of ligand in the effector site induces structural changes that spread through much of the subunit, but cause only minor changes in the active site . It is not clear why the crystals should contain the enzyme in a high-affinity conformation, which presumably represents only a small fraction of the molecules in the crystallizing solution . However, this structure does identify the conformational changes involved in binding of the allosteric effectors. J Immunol, 1989 Jun 15, 142(12), 4507 - 13 Linkage analysis of the Bcg gene on mouse chromosome 1 . Identification of a tightly linked marker; Schurr E et al.; We have mapped and determined the gene order of five cloned genes in the vicinity of the murine host resistance gene Bcg on mouse chromosome 1 . For this, we have used a RFLP-type analysis in panels of 43 recombinant inbred strains, 3 congenic mouse strains, and 186 segregating backcross progeny derived from inbred strains of Bcgr and Bcgs genotypes . The Bcg alleles of segregating animals were established by in vivo infection with Mycobacterium bovis (Bacillus Calmette-Guerin) strain Montreal . Genomic DNA prepared from progenitor mouse strains was isolated, digested with restriction endonucleases, and analyzed by Southern blotting to identify strain-specific RFLP for each DNA marker tested . Among a number of DNA markers tested, Len2, Fn, Vil, Alpi, and Achrg were found to co-segregate with Bcg in mouse strains congenic for this locus . Detailed segregation analysis of the five markers and Bcg showed that Vil was extremely close to Bcg with no recombinant identified, whereas Fn and Len2 were located 4.5 and 9 cM proximal of Bcg, respectively . Alpi and Achrg mapped 5 and 5.5 cM distal from Bcg, respectively . Pedigree analysis in the recombinant inbred strains and backcross animals indicated the gene order: centromere-Len2-Fn-Vil,Bcg-Alpi-Achrg . The tightly linked Vil marker can now be used as an entry point in recombinant genomic DNA libraries to clone sequences overlapping Bcg . This group of five genes flanking Bcg on mouse chromosome 1 is precisely conserved on the telomeric end of the long arm of human chromosome 2q . Our results suggest that a likely location for a putative human homologue to the murine host resistance gene Bcg is the long arm of human chromosome 2 (2q32-qter). J Clin Microbiol, 1989 Jun, 27(6), 1395 - 6 Bacteremia and infection of a hip prosthesis caused by Bacillus alvei; Reboli AC et al.; Of the 34 Bacillus species described, 10 have been reported to cause infection in humans and 6 are insect pathogens . We report a case of an infected prosthetic hip caused by Bacillus alvei, only the third documented case of human infection with this organism. Br J Urol, 1989 Jun, 63(6), 610 - 5 Intravesical Bacillus Calmette-Guérin treatment for superficial bladder tumours; Shinka T et al.; Intravesical instillations of Tokyo 172 strain BCG were given to 56 patients with superficial bladder cancer during the 24-month period after transurethral tumour resection as a prophylaxis against tumour recurrence . The recurrence rate of tumours was compared with that of historical controls . Results were estimated by the person-years method and there were statistically significant decreases in recurrent tumours following BCG therapy . Our results suggest that the intravesical Tokyo 172 strain BCG is effective and safe as a prophylaxis against the recurrence of superficial bladder tumours. Minerva Ginecol, 1989 Jun, 41(6), 287 - 90 {Non-specific vaginitis and topical treatment . Comparison of flunoxaprofen and benzydamine}; Cecchini G et al.; The efficacy and tolerability of a new NSAID-flunoxaprofen-have been evaluated in patients suffering from non specific vaginitis, by topical application for 20 days (vaginal washings with water solution of the preparation) . The activity of flunoxaprofen has been compared with that of benzidamine with regard to normalization of bacterial vaginal flora, taking into consideration the increase of Doderlein bacillus . A remarkable significant improvement of all the symptoms has been observed in the group of patients treated with flunoxaprofen with respect to that receiving benzidamine; moreover 57.9% of the subjects treated with flunoxaprofen showed a significant increase of Doderlein bacillus while in the benzidamine group the percentage reached a value of 11.8% . Flunoxaprofen may be considered a useful and active tool for the topical treatment of non specific vaginal diseases. Microbiol Rev, 1989 Jun, 53(2), 242 - 55 Insecticidal crystal proteins of Bacillus thuringiensis; Hofte H et al.; A classification for crystal protein genes of Bacillus thuringiensis is presented . Criteria used are the insecticidal spectra and the amino acid sequences of the encoded proteins . Fourteen genes are distinguished, encoding proteins active against either Lepidoptera (cryI), Lepidoptera and Diptera (cryII), Coleoptera (cryIII), or Diptera (cryIV) . One gene, cytA, encodes a general cytolytic protein and shows no structural similarities with the other genes . Toxicity studies with single purified proteins demonstrated that every described crystal protein is characterized by a highly specific, and sometimes very restricted, insect host spectrum . Comparison of the deduced amino acid sequences reveals sequence elements which are conserved for Cry proteins . The expression of crystal protein genes is affected by a number of factors . Recently, two distinct sigma subunits regulating transcription during different stages of sporulation have been identified, as well as a protein regulating the expression of a crystal protein at a posttranslational level . Studies on the biochemical mechanisms of toxicity suggest that B . thuringiensis crystal proteins induce the formation of pores in membranes of susceptible cells . In vitro binding studies with radiolabeled toxins demonstrated a strong correlation between the specificity of B . thuringiensis toxins and the interaction with specific binding sites on the insect midgut epithelium . The expression of B . thuringiensis crystal proteins in plant-associated microorganisms and in transgenic plants has been reported . These approaches are potentially powerful strategies for the protection of agriculturally important crops against insect damage. J Biol Response Mod, 1989 Jun, 8(3), 262 - 77 Hyperthermic modulation of tumor necrosis factor-dependent monocyte/macrophage tumor cytotoxicity in vitro; Klostergaard J et al.; Tumor necrosis factor (TNF) production by human peripheral blood monocytes and murine bacillus Calmette-Guerin-activated peritoneal macrophages was strongly influenced by acute hyperthermia . If hyperthermia was administered simultaneously with or preceding lipopolysaccharide triggering, production was severely ablated by 42 degrees and 43 degrees C treatments; however, if triggering preceded heating by at least 90 min, production was either unaffected or markedly enhanced . A somewhat similar pattern was reflected with chronic heating . TNF production by murine macrophages was inhibited with 39 degrees C heating and completely blocked by 40.5 degrees C treatment, if triggering coincided with the initiation of hyperthermia . However, augmentation of production occurred with either of these temperatures if triggering preceded hyperthermia by as little as 90 min . Human monocytes demonstrated greater resistance to the deleterious effects of coincident triggering and heating with respect to TNF secretion than the rodent effectors, but the response was otherwise very similar . The TNF-sensitive phenotype of the L929 cell could be augmented by chronic or acute hyperthermia, markedly so with a 43 degrees C treatment . The TNF-resistant phenotype of the EMT-6 cell could be reversed by chronic heating at 40.5 degrees C, or by acute heating at 43 degrees C, but only if the latter followed TNF treatment . These results reflect important regulatory controls of TNF production and responses in tumor cells which are susceptible to hyperthermia manipulation. Int J Lepr Other Mycobact Dis, 1989 Jun, 57(2), 451 - 7 Results of a modified WHO regimen in highly bacilliferous BL/LL patients; Katoch K et al.; A regimen consisting of 600 mg of rifampin once a month, 100 mg of clofazimine on alternate days, and 100 mg of dapsone daily was used in 56 untreated, highly bacillated borderline lepromatous/lepromatous (BL/LL) patients with an average bacterial index (BI) of 4.45 . Treatment was continued until skin-smear negativity . After 2 years of therapy, none of the patients had become smear negative and the average BI was 2.56 . There was no growth on inoculation of skin-tissue biopsies in the normal mouse foot pad after 6 months of therapy . Bacillemia was still detectable in 11/50 patients, and significant ATP levels were detected in Mycobacterium leprae from skin-tissue biopsies in 16% of the cases . After 3 years of therapy, three patients had become smear negative . The average BI was 1.30 . None of the patients had detectable bacillemia, and 5% of the cases showed detectable ATP levels in M . leprae from tissue biopsies . After 4 years of therapy, 41.7% of the patients had become smear negative . The average BI was 0.66, and no ATP was detected in any of the purified bacillary suspensions . The fall in BI was accelerated, and more patients on continued treatment became negative earlier compared to those having treatment for a limited duration, as reported by others. J Urol, 1989 Jun, 141(6), 1449 - 53 Class I and class II HLA antigen expression by transitional cell carcinoma of the bladder: correlation with T-cell infiltration and BCG treatment; Stefanini GF et al.; HLA class I and II glycoproteins from transitional cell carcinoma (TCC) and from perineoplastic and healthy vesical mucosa were characterized together with infiltrating cells by means of immunochemistry using specific monoclonal antibodies on frozen sections obtained during resection or radical cystectomy . Specimens were taken from 11 patients with TCC and five with healthy bladder mucosa . Four patients with TCC and four with healthy mucosa had been previously treated with a course of intravesical bacillus Calmette-Guerin (BCG) . Ten out of 11 TCC samples expressed class I glycoproteins with a membrane pattern (diffuse in seven, focal in three) as normal epithelial cells from either controls or perineoplastic bladder . Interestingly, eight out of 11 TCC samples expressed class II antigens on their membrane that were also present in six cases in the perineoplastic tissue while the epithelial cells from four out of five normal bladders were completely negative . The epithelial display of class II antigens in the non-neoplastic areas and in the normal bladder correlates (p less than 0.001) with the degree of cellular infiltrate while such a relationship was not found between the HLA II expression of neoplastic cells and the infiltrate . BCG treatment was associated with a higher amount of inflammatory cells, prevalently T "activated" cells (CD5+,DR+), with a CD4/CD8 ratio always greater than 1 . In the light of the role played by HLA glycoproteins in immune mechanisms, these results could help explain the positive action of BCG and the relative immunosensitivity of TCC. J Bacteriol, 1989 Jun, 171(6), 3568 - 71 Formation of crystals of the insecticidal proteins of Bacillus thuringiensis subsp . aizawai IPL7 in Escherichia coli; Oeda K et al.; Escherichia coli JM103 cells harboring expression plasmid pTB1 or pKC6 synthesized the 130- and 135-kilodalton insecticidal proteins, respectively, of Bacillus thuringiensis subsp . aizawai IPL7, and both products accumulated as cytoplasmic inclusion bodies . Amorphous inclusions which contained contaminating proteins, together with the corresponding insecticidal proteins, were formed in cultures at 37 degrees C, but bipyramidal crystals practically free of contaminants were observed at 30 degrees C . Although 9.8% of the amino acids were substituted between these two proteins, both protein crystals had the same shape as those of the parental B . thuringiensis strain, which produced both proteins. J Bacteriol, 1989 Jun, 171(6), 3060 - 7 Purification and properties of a 28-kilodalton hemolytic and mosquitocidal protein toxin of Bacillus thuringiensis subsp . darmstadiensis 73-E10-2; Drobniewski FA et al.; The mosquitocidal crystal of Bacillus thuringiensis subsp . darmstadiensis 73-E10-2 was purified, bioassayed against third-instar Aedes aegypti larvae (50% lethal concentration, 7.5 micrograms/ml), and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealing polypeptides of 125, 50, 47, and 28 kilodaltons (kDa) . When solubilized and proteolytically activated by insect gut proteases or proteinase K, the crystal was cytotoxic to insect and mammalian cells in vitro and was hemolytic . By using nondenaturing polyacrylamide gel electrophoresis, a polypeptide of 23 kDa, derived from the 28-kDa protoxin, was identified which was hemolytic and cytotoxic to Aedes albopictus, A . aegypti, and Choristoneura fumiferana CF1 insect cell lines . The 23-kDa polypeptide was purified by ion-exchange chromatography and gave 50% lethal dose values of 3.8, 3.3, and 6.9 micrograms/ml against A . albopictus, A . aegypti, and C . fumiferana CF1 cells lines, respectively . Cytotoxicity in vitro was both dose and temperature dependent, with a sigmoidal dose-response curve . The cytotoxicity of the 23-kDa toxin and the solubilized and proteolytically activated delta-endotoxin was inhibited by a range of phospholipids containing unsaturated fatty acids and by triglyceride and diglyceride dispersions . An interaction with membrane phospholipids appears important for toxicity . Polyclonal antisera prepared against the 23-kDa polypeptide did not cross-react with polypeptides in the native crystals of four other mosquitocidal strains. J Egypt Soc Parasitol, 1989 Jun, 19(1), 195 - 203 Effect of Bacillus sphaericus and Bacillus thuringiensis on acid-phosphatase activity of mosquito larvae, Culex pipiens and Aedes caspius; Hussein MA et al.; The use of Bacillus sphaericus and B . thuringiensis H-14 form one of the important group of biological control agent against mosquito larvae . Acid phosphatase enzyme plays a significant role in determining susceptibility of mosquito larvae to both bacterial species . Biochemical assay showed activation in acid phosphatase in Culex pipiens and Aedes caspius treated with B . thuringiensis . Variation in acid phosphatase activity occurred in both mosquitoes treated with B . sphaericus, while there was no change in acid phosphatase activity in A . caspius . An obvious increase in activity in C . pipiens treated with the same bacteria was observed. Mol Gen Mikrobiol Virusol, 1989 Jun, (6), 42 - 5 {New producers of site-specific endonucleases from microorganisms of the Bacillus genus}; Kramarov VM et al.; 52 strains of Bacillus generum have been tested for production of site-specific endonucleases . The sequence recognized by the enzyme was determined for 23 enzymes, the cleavage site inside the sequence was determined for 5 enzymes . All the enzymes under study were found to be isomers of the known enzymes . The selected strains are peculiar for the high level of site-specific endonucleases content and may be used as producents of the enzymes. Proc Natl Acad Sci U S A, 1989 Jun, 86(11), 4037 - 41 Location of the Bombyx mori specificity domain on a Bacillus thuringiensis delta-endotoxin protein; Ge AZ et al.; Bacillus thuringiensis produces different types of insecticidal crystal proteins (ICPs) or delta-endotoxins . In an effort to identify the insect specificity of ICP toxins, two icp genes were cloned into the Escherichia coli expression vector pKK223-3, and bioassays were performed with purified crystals . The type A protein {from an icpA1, or 4.5-kilobase (kb) gene, from B . thuringiensis var . kurstaki HD-1} was found to be 400 times more active against Bombyx mori than type C protein (from an icpC73, or 6.6-kb gene, from B . thuringiensis var . kurstaki HD-244) . The type C protein was 9 times more active against Trichoplusia ni than the type A protein, while both have similar activity against Manduca sexta . To locate the specificity domain of the type A protein for B . mori, site-directed mutagenesis was used to introduce or remove restriction enzyme sites, facilitating the exchange of regions of the two genes . The hybrid genes were overexpressed, and purified ICP was used in bioassays . The B . mori specificity domain for the ICP A toxin is located in the amino-terminal portion of the hypervariable region between amino acids 332 and 450. Tubercle, 1989 Jun, 70(2), 139 - 41 Drug resistant tuberculous meningitis in the Philippines: report of a case; Watt G et al.; A fatal case of tuberculous meningitis caused by a multiply-resistant tubercle bacillus is described, the first such case from Southeast Asia . Increased efforts to isolate Mycobacterium tuberculosis from the cerebrospinal fluid and determine the extent and pattern of drug resistance are necessary if the high mortality from this disease is to be reduced. Chin Med J (Engl), 1989 Jun, 102(6), 464 - 8 Studies on the efficacy and persistence of the microbial agent bacillus sphaericus against larvae of culex pipiens pallens; Zhen TM et al.; This paper evaluates the efficacy of the bacterial larvicide bacillus sphaericus strains BS-10 and C3-41, which are isolated in China, as well as BS2362 against larvae of Culex pipiens pallens in laboratory and under field conditions . The results indicate that C3-41 has the highest toxicity with LC50 at 0.0057 ppm and the bacterial formulations are more effective in light polluted water than in heavy polluted one, but the action of B . sphaericus could persist longer in more polluted water. Genetika, 1989 Jun, 25(6), 1013 - 20 {Transduction ability of mutants of phage CP51, virulent for bacteria of the Bacillus cereus group}; Koretskaia NG et al.; The virulent phage CP51 used usually to transfer chromosomal and plasmid markers between bacteria of the Bacillus cereus group was treated with N-methyl-N'-nitro-N-nitroguanidine . Mutants with reduced viability and ts-mutants were isolated . Some of the mutants were found to have an increased efficiency of transduction and allow to simplify the process . Transfer frequencies of the plasmid pBC16 by the phages CP51-26 and CP51-4-59 were 5 x 10(-4) per plaque-forming unit and 4-5 x 10(-3) per bacterial cell, respectively . Possibilities of further increasing the transduction efficiency of Bacillus thuringiensis genetic material using phage CP51 mutants are discussed. Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi, 1989 Jun, 5(2), 100 - 2, 157 {Determination of blood endotoxin in severely burned patients and its clinical significance}; Zhang YP; Since October 1985 to June 1987, in 12 severely burned patients with endotoxemia was observed by using a quantitative endotoxin assay of limulus amoebocyte lysate (LAL) test with a chromogenic substrate . Among the 12 patients, 4 died and 8 survived . The average age of dead group was 31.8 years (19-45 years), mean TBSA was 63% (58-80%) and mean I 18.5% . The survival group average age of survival group was 27.5 years (18-39 years), mean TBSA was 58% (18-85%) and mean I 24.4% (6-56%) . The plasma endotoxin concentrations of burn patients in dead group were 105-571 pg/ml, significantly higher than that of survival group (30-240 pg/ml) and healthy human (6.44 +/- 1.96 pg/ml) . It was found that the increase of endotoxemia was closely related to burn wound sepsis, positive of blood culture, systemic disseminated septicaemia . Systemic use of sensitive antibiotics may increase the level of blood endotoxin in severe gram-negative bacillus infection . Polymixin-B is an exception. Ann Soc Belg Med Trop, 1989 Jun, 69(2), 143 - 7 {Initial resistance to streptomycin, isoniazid, thiacetazone, rifampicin and ethambutol in bacilliferous tuberculosis in Maniema, Zaire}; De Caluwe P et al.; Initial resistance to streptomycin, isoniazid, thiacetazone, rifampicin and ethambutol was tested in 102 patients with bacilliferous tuberculosis in Kalima, a rural area in eastern Zaire . The initial resistance for at least one of these tuberculostatics was 43% . The highest resistance recorded was for streptomycin (31%) . No resistance to rifampicin or ethambutol was found . The practical interest of these findings is discussed. Lepr Rev, 1989 Jun, 60(2), 94 - 101 Testicular dysfunction in leprosy: relationships of FSH, LH and testosterone to disease classification, activity and duration; Levis WR et al.; Luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone levels were determined by radioimmunoassay (RIA) in leprosy patients and analysed for effect of disease classification, disease activity and duration of disease . LH and FSH levels were found to be significantly elevated in lepromatous patients compared to borderline-lepromatous, midborderline and borderline-tuberculoid patients . A positive correlation was seen between LH and FSH and a negative correlation was seen between testosterone and both LH and FSH . No correlation was seen between hormone levels and measures of disease activity: bacillary index and IgM to phenolic glycolipid I, a Mycobacterium leprae antigen . A significant correlation was seen between duration of disease and FSH when age was taken into account, indicating that testicular dysfunction is probably cumulative and irreversible . It is recommended that LL patients be routinely screened for hypogonadism using FSH, LH and testosterone levels. Appl Environ Microbiol, 1989 Jun, 55(6), 1649 - 52 Transformation of Bacillus cereus vegetative cells by electroporation; Belliveau BH et al.; Transformation of untreated vegetative cells of Bacillus cereus 569 with plasmid pC194 (1.8 megadaltons) by high-voltage electroporation resulted in a maximum of 2 x 10(-5) transformants per viable cell . Transformation of a 130-megadalton plasmid occurred at a comparable frequency . The method was simple, rapid, and yielded transformant colonies in 14 to 24 h . Transformation was obtained with unpurified total plasmid DNA. South Med J, 1989 Jun, 82(6), 705 - 9 Clinical significance of Bacillus species isolated from blood cultures; Weber DJ et al.; To determine the clinical significance of blood isolates of Bacillus, we reviewed all blood cultures obtained at North Carolina Memorial Hospital between 1981 and 1985 . Over the five-year study period the number of patients (incidence per 10,000 hospital admissions) from whom Bacillus was isolated increased from 4.97 in 1981 to 12.5 in 1985 . The incidence per 1,000 blood cultures also increased from 1.12 in 1981 to 2.33 in 1985 . Review of the medical records of 78 of the 95 patients (82%) with positive cultures allowed retrospective classification of five isolates (6.4%) as clinically significant, 33 isolates (42.3%) as possibly significant, and 40 isolates (51.3%) as nonsignificant . Underlying diseases in patients with clinically significant Bacillus bacteremia included burn trauma in two, leukemia in one, carcinoma in one, and gastrointestinal hemorrhage in one . All isolates judged to be clinically significant and the majority of possibly significant isolates were B cereus . We conclude that the isolation of Bacillus species from blood cultures is clinically significant in 5% to 10% of cases, that the incidence of Bacillus bacteremia is increasing, and that burn trauma should be added to the list of conditions known to predispose to clinically significant Bacillus bacteremia. J Gen Microbiol, 1989 Jun, 135 ( Pt 6), 1521 - 8 Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus; Kuriki T et al.; The gene (nplT) for a new type of pullulan-hydrolysing enzyme, neopullulanase, from Bacillus stearothermophilus TRS40 was sequenced . The DNA sequence revealed only one large open reading frame, composed of 1764 bases and 588 amino acid residues (Mr 69144) . Although the thermostable neopullulanase contained eight cysteine residues, they did not provide conformational stability by disulphide bonds . A comparison was made of the amino acid sequences of alpha-amylase, neopullulanase, isoamylase, pullulanase and cyclodextrin glucanotransferase . All the enzymes examined contained four highly conserved regions which probably constitute the active centres of the enzymes . The amino acid residues required for the specificity of neopullulanase are compared with those of alpha-amylase and other amylolytic enzymes. Biochem Biophys Res Commun, 1989 May 30, 161(1), 59 - 63 On the effect on specificity of Thr246----Gly mutation in L-lactate dehydrogenase of Bacillus sterothermophilus; Bur D et al.; The function of the amino acid Thr246 in L-lactate dehydrogenase from Bacillus stearothermophilus has been investigated by site-directed replacement with glycine . Kinetic experiments with a number of 2-oxo acids showed strongly reduced activity for the mutated enzyme . However, the mutant enzyme shows a relative preference for the large hydrophobic sidechains of alpha-keto acids and an even higher specific activity than the wild-type lactate dehydrogenase for the polar oxaloacetate substrate . Graphic analyses indicate that the loss of one hydrogen bond, or intrusion of water into the active site, might be responsible for the reduced activity . The kinetic results suggest that the binding modes of bulky hydrophobic or polar substrates compensate to some degree for the partially disrupted active site. Biochem J, 1989 May 15, 260(1), 87 - 91 Facile preparation and characterization of the toxin from Bacillus thuringiensis var . kurstaki; Bietlot H et al.; We report a simple three-step method of generating a homogeneous toxic fragment (toxin) in high yield from B . thuringiensis var . kurstaki . Purified crystals were digested with trypsin at pH 10.5, followed by (NH4)2SO4 precipitation and dialysis . For the HD73 strain the preparation is toxic to eastern-spruce-budworm (Choristoneura fuminiferana) larvae . It gives a single 66 kDa band on polyacrylamide-gel electrophoresis and a single band with an isoelectric point of 5.5 on an isoelectric-focusing gel . A single isoleucine N-terminus was detected, and the first 20 amino acids were found to be identical with those predicted from the gene nucleotide sequence . A single lysine C-terminus was detected, and the amino acid composition was in excellent agreement with tryptic cleavages at arginine-28 and lysine-623 of the protoxin . Raman spectroscopic analysis gave values of 20% alpha-helix, 35% beta-sheet and 45% unordered structure . The resistance of the toxin to most proteinases and its susceptibility to proteolysis by papain and Pronases indicates a compact multidomain structure. J Biol Chem, 1989 May 5, 264(13), 7447 - 54 New-found phenolic glycolipids in Mycobacterium bovis BCG . Presence of a diglycosylated glycolipid; Vercellone A et al.; A crude phenolic glycolipid extract from Mycobacterium bovis bacille Calmette-Guerin (BCG) was fractionated until homogeneity at the intact level into four phenolic glycolipids called B, B-1, B-2, and B-3 according to their polarity . The apolar one, which is the most abundant was assigned to the well-known mycoside B . The B-2 and B-3 phenolic glycolipids were purified by direct-phase high performance liquid chromatography using a 5 micron Spherisorb column but were only recovered in small amounts (3 mg) . A linear gradient of 0-20% methanol in chloroform was used . The B-1, B-2, and B-3 glycolipids were subjected to suitable modern anal