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| 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 analytical techniques selected for their potential to elucidate the structure at the intact level . Desorption chemical ionization-mass spectrometry allowed the molecular mass of B-3 to be determined as 1652 Da for the major homolog establishing the molecular formula as C103H192O14 . Thus, the B-3 polar phenolic glycolipid contained two deoxyhexoses, one molecule of phenolphthiocerol esterified by two molecules of mycocerosic acid . Using two-dimensional 1H NMR (correlated chemical shift and nuclear Overhauser effect spectroscopy) at the intact level the B-3 oligosaccharide structure was determined as an alpha-L-Rhap-(1----3)-2-O-Me-alpha-L-Rhap . This is the first report of a diglycosylated phenolic glycolipid in a nonpathogenic mycobacteria . The disaccharide unit, the antigenic determinant, appears to be characteristic of M . bovis BCG . This polar glycolipid B-3 and the apolar ones, B-1 and B-2, were reactive in enzyme-linked immunosorbent assay against serum from rabbit hyperimmunized with M . bovis BCG. Biochemistry, 1989 May 2, 28(9), 3843 - 50 Kinetic characterization of the recombinant ribonuclease from Bacillus amyloliquefaciens (barnase) and investigation of key residues in catalysis by site-directed mutagenesis; Mossakowska DE et al.; Barnase, the ribonuclease from Bacillus amyloliquefaciens, has been cloned and expressed in Escherichia coli {Hartley, R . W . (1988) J . Mol . Biol . 202, 913-915}, thus enabling the overproduction and site-directed mutagenesis of one of the smallest enzymes (Mr equals 12,382) . As barnase is also composed of just a single polypeptide chain with no disulfide bridges and has a reversible folding transition, it affords a fine system for studying protein folding and design . We show here that the recombinant enzyme has properties identical with those of the authentic enzyme, characterize the basic kinetics and specificity of the enzyme, and, using site-directed mutagenesis, identify key residues involved in catalysis to provide evidence that supports the classic ribonuclease mechanism . The wild-type enzyme catalyzes the hydrolysis of dinucleotides of structure GpN . There is a prime requirement for G and a preference for A greater than G greater than C greater than U for N . The pH-activity curve for the transesterification step of dinucleotides is bell shaped with an optimum for kcat/KM and kcat at about pH 5 . The enzyme is far more active toward long RNA molecules, and the pH optimum for kcat is at 8.5 . The activity of barnase toward dinucleotide substrates is about 0.5% of that of the highly homologous T1 nuclease at pH 5.9, but barnase is twice as active as T1 toward RNA at pH 8.5 . There must be important subsite interactions that contribute to catalysis in barnase in addition to those immediately on either side of the scissile bond.(ABSTRACT TRUNCATED AT 250 WORDS) Mikrobiol Zh, 1989 May-Jun, 51(3), 63 - 5 {L-forms of aerobic spore-forming bacteria isolated from urologic patients}; Rudenko AV et al.; New data are presented on the ability of different aerobic spore-forming bacteria isolated from the organism of urological patients to produce L-forms of these microorganisms in the presence of penicillin and ampicillin . Bacillus cereus is shown to be the most resistant to these antibiotics. Mikrobiol Zh, 1989 May-Jun, 51(3), 3 - 6 {Optimization of the nutrient medium composition for directed biosynthesis of Bacillus mesentericus lectins}; Simonenko IA; The medium has been optimized in order to increase the lectin-producing activity of Bacillus mesentericus . When a producer grew on the optimized medium, a 4-8-fold increase of the hemagglutination test titres and a 5.6-fold increase (as compared with the initial medium) of the specific lectin activity of the culture liquid was attained. Ann Rheum Dis, 1989 May, 48(5), 432 - 4 Arthritis associated with adjuvant mycobacterial treatment for carcinoma of the bladder; Hughes RA et al.; A patient who developed an inflammatory polyarthritis following intravesical administration of bacillus Calmette-Guerin (BCG) used in the treatment of bladder cancer is described . An inflammatory synovitis comprising predominantly T lymphocytes was demonstrated on synovial biopsy . The synovitis resolved spontaneously within 14 days in this 'human model' of adjuvant arthritis. Proc Natl Acad Sci U S A, 1989 May, 86(10), 3589 - 93 Use of Bacillus brevis for efficient synthesis and secretion of human epidermal growth factor; Yamagata H et al.; Using previously isolated Bacillus brevis strains that secrete large amounts of proteins but little protease into the medium, we have developed a host-vector system for very efficient synthesis and secretion of heterologous proteins . The multiple promoters and the signal-peptide-coding region of the MWP gene, a structural gene for one of the major cell wall proteins of B . brevis strain 47, were used to construct expression-secretion vectors . With this system, a synthetic gene for human epidermal growth factor (hEGF) was expressed efficiently and a large amount (0.24 g per liter of culture) of mature hEGF was secreted into the medium . hEGF purified from the culture supernatant had the same NH2-terminal amino acid sequence, COOH-terminal amino acid, and amino acid composition as natural hEGF, and it was fully active in biological assays . These results, in combination with previous results, showed that mammalian proteins can be produced in active form 10-100 times more efficiently in B . brevis than has been reported in other systems. Indian J Med Res, 1989 May, 89, 150 - 7 Use of eluates of filter paper blood spots in ELISA for the serodiagnosis of leprosy; Dhandayuthapani S et al.; Blood samples were collected from 59 leprosy patients and 35 normal healthy subjects by veni-puncture and finger prick methods to obtain serum samples and blood spots on filter paper respectively . The serum samples at 1:300 dilutions and the eluates of dried blood spots at 1:40, 1:80, 1:160 and 1:320 dilutions were applied in ELISA to measure the antibody levels (IgM) against synthetic ND-O-BSA antigen . The antibody levels were found to be high in the multibacillary leprosy patients than the pauci-bacillary patients irrespective of whether serum samples or eluates were used . The OD values obtained at 1:160 dilution of the eluates were equivalent to that of values obtained at 1:300 dilution of the serum samples . The positivities differ in different dilution of the eluates, showing the highest in the 1:40 dilution and the lowest in the 1:320 dilution. Eur J Biochem, 1989 May 1, 181(2), 443 - 51 Molecular and immunological characterization of plastid and cytosolic pyruvate kinase isozymes from castor-oil-plant endosperm and leaf; Plaxton WC; 1 . Monospecific antiserum was raised in rabbits to homogeneous cytosolic pyruvate kinase isolated from 5-day-old germinating endosperm of the castor oil plant, Ricinus communis . An earlier study demonstrated that the purified enzyme is putatively heterotetrameric, composed of two subunits which migrate as 57-kDa and 56-kDa proteins upon sodium dodecyl sulfate/polyacrylamide gel electrophoresis {Plaxton, W . C . (1988) Plant Physiol . (Bethesda) 86, 1065-1069} . Both proteins were detected on Western blots of extracts prepared under denaturing conditions from 4-8-day-old, but not 0-3-day-old, germinating-endosperm tissue . This suggests that both subunits exist in vivo, and that the large increase in pyruvate kinase activity which occurs around the fourth day of germination is due to an increase in pyruvate kinase concentration . 2 . The cytosolic and plastidic pyruvate kinase isozymes (termed PKc and PKp, respectively) from castor-oil-plant developing endosperm and expanding leaf tissue were separated by anion-exchange chromatography on Q-Sepharose . The antigenic reaction of the partially purified enzyme preparations to rabbit polyclonal antibodies raised against homogeneous germinating-castor-bean PKc was tested by immunoprecipitation and Western blotting . Although developing-endosperm and leaf PKc appeared to be antigenically very similar to germinating-endosperm PKc, they differed from the heterotetrameric germinating-endosperm enzyme by being composed of a single type of subunit with a molecular mass of about 56 kDa . No cross-reactivity of the PKc antibodies was observed with either developing-endosperm or leaf PKp, nor with rabbit muscle or Bacillus stearothermophilus pyruvate kinase . Conversely, none of the castor-oil-plant pyruvate kinase preparations showed significant cross-reactivity with antibodies raised against purified yeast or rabbit muscle pyruvate kinases . 3 . To investigate the structural relationship between the two germinating-endosperm-PKc subunits, each polypeptide was characterized by amino acid composition analysis and peptide mapping by CNBr fragmentation . The amino acid compositions and CNBr cleavage patterns of the two subunits were similar, but not identical, suggesting that these polypeptides are related, but distinct, proteins . Mild tryptic attack of native enzyme led to an approximate 6-kDa reduction in the apparent molecular mass of both subunits, further indicating sequence similarity between the two polypeptides . 4 . Native molecular masses of the various castor-oil-plant pyruvate kinases were estimated by Superose-6 gel-filtration chromatography.(ABSTRACT TRUNCATED AT 400 WORDS) J Bacteriol, 1989 May, 171(5), 2835 - 41 Function of alpha-D-glucosyl monophosphorylpolyprenol in biosynthesis of cell wall teichoic acids in Bacillus coagulans; Shimada A et al.; D-{alpha-14C}}glucosyl phosphorylpolyprenol ({ 14C}Glc-P-prenol) was formed from UDP-D-{14C}glucose in each of the membrane systems obtained from Bacillus coagulans AHU 1631 and AHU 1634 and two Bacillus megaterium strains . Membranes of these B . coagulans strains, which possess beta-D-glucosyl branches on the repeating units in their major cell wall teichoic acids, were shown to catalyze the transfer of the glucose residue from {14C}Glc-P-prenol to endogenous polymer . On the other hand, membranes of B . coagulans AHU 1366, which has no glucose substituents in the cell wall teichoic acid, exhibited neither {14C}Glc-P-prenol synthetase activity nor the activity of transferring glucose from {14C}Glc-P-prenol to endogenous acceptor . The enzyme which catalyzes the polymer glycosylation in the former two B . coagulans strains was most active at pH 5.5 and in the presence of the Mg2+ ion . The apparent Km for {14C}Glc-P-prenol was 0.6 microM . Hydrogen fluoride hydrolysis of the {14C}glucose-linked polymer product yielded a major fragment identical to D-galactosyl-alpha(1----2)(D-glucosyl-beta(1----1/3)) glycerol, the dephosphorylated repeating unit in the major cell wall teichoic acids of these B . coagulans strains . This result, together with the behavior of the radioactive polymer in chromatography on Sepharose CL-6B, DEAE-Sephacel, and Octyl-Sepharose CL-4B, led to the conclusion that {14C}Glc-P-prenol serves as an intermediate in the formation of beta-D-glucosyl branches on the polymer chains of cell wall teichoic acids in B . coagulans. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1989 May, 22(2), 97 - 104 Enzyme-linked immunosorbent assay with BCG sonicate antigen for diagnostic potential of mycobacterial infection in Taiwan; Wang CR et al.; The diagnostic potential of enzyme-linked immunosorbent assay (ELISA) with Bacillus Calmette-Guerin (BCG) sonicate antigen for detection of mycobacterial infections, including pulmonary tuberculosis and leprosy, has been evaluated in Taiwan area . One hundred blood samples were collected from 74 active and 26 inactive pulmonary tuberculosis patients of the Taiwan Provincial Tuberculosis Control Bureau . Another 50 samples were collected from 24 lepromatous, 23 tuberculoid and 3 borderline leprosy patients at the Taiwan Provincial Lo-Sheng Leprosarium . The IgG anti-BCG sonicate levels were compared among patients with tuberculosis, active or inactive, patients with leprosy, and healthy individuals . Patients with tuberculosis, both active and inactive, and those with leprosy had higher BCG sonicate antibody levels and frequencies above cutoff value than healthy subjects (for BCG-sonicate antibody, p less than 0.05 for inactive tuberculosis, p less than 0.0001 for others; for frequency above cutoff, p less than 0.001 for inactive tuberculosis, p less than 0.0001 for others) . Among the three groups of patients, significant differences were noted in anti-BCG sonicate antibody level, and there was no difference of frequency above the cutoff value (i.e . inactive tuberculosis had lower level than active tuberculosis and leprosy, p less than 0.005) . In conclusion, ELISA with BCG sonicate antigen could serve as a diagnostic aid for mycobacterial infection in Taiwan . However, it was not possible to differentiate disease activity in tuberculosis, or to discriminate infectious species of mycobacteria . Purified mycobacterial antigens should be tried in further research to improve the specificity and sensitivity in serodiagnosis with ELISA. Acta Otorrinolaringol Esp, 1989 May-Jun, 40(3), 189 - 94 {Tuberculosis in current clinical otorhinolaryngology}; Abdel-Hadi M et al.; 39 cases of ENT tuberculosis diagnosed in the last 15 years are studied . We review the annual incidence, the distribution by age and sex and the different found localization: oropharyngeal, laryngeal cervical ganglionar, salivary and otical ones . We analyse the concomitance with tuberculous pulmonary lesions and the results of diagnostic explorations: Mantoux, bacilloscopy and biopsy . We discuss the method, diagnostic difficulties, therapeutic behaviour and the consequences of misdiagnosis. Q J Med, 1989 May, 71(265), 407 - 16 Fungal peritonitis complicating peritoneal dialysis: report of 27 cases and review of treatment; Cheng IK et al.; The clinical features, treatment and outcome of 27 cases of fungal peritonitis were studied . Twenty-one cases occurred in patients receiving CAPD and six in patients on intermittent peritoneal dialysis . Twenty-five cases were due to Candida spp., one was due to Trichosporon spp . and in one, both Candida and Trichosporon and an unidentified acid-fast bacillus were isolated . Clinical features of fungal peritonitis and bacterial peritonitis were the same . A direct comparison with patients without fungal peritonitis failed to reveal an increased incidence of diabetes mellitus . However, a history of recent bacterial peritonitis and antibiotic treatment was frequently obtained . We found that the combination of oral ketoconazole and intraperitoneal miconazole is successful in treating fungal peritonitis complicating peritoneal dialysis but catheter removal and replacement is often necessary . Analysis of the relationship between clinical outcome and various treatment strategies in cases reported in the literature and in our own showed that an initial trial of antifungal drugs consisting of oral ketoconazole and i.p . 5-fluorocytosine or miconazole is warranted in most cases before contemplating catheter removal. Mol Microbiol, 1989 May, 3(5), 621 - 6 Molecular characterization and sequence of phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis; Lechner M et al.; The gene encoding monophosphatidylinositol inositol phosphohydrolase (PI-specific phospholipase C, PI-PLC) of Bacillus thuringiensis was cloned in Staphylococcus carnosus TM300 . The complete coding region comprises 987 base pairs corresponding to a precursor protein of 329 amino acids (molecular weight, 38,095) . The NH2-terminal sequence of the purified enzyme from Escherichia coli indicated that the mature PI-PLC consists of 299 amino acid residues with a molecular weight of 34,586 . Polyacrylamide gel electrophoresis revealed the same molecular weight for the purified enzyme isolated from the DNA-donor strain of B . thuringiensis and from the E . coli clone . By computer analysis, the secondary structure was predicted . The enzyme from the E . coli recombinant shows no activity on other phospholipids and sphingomyelin . The cleaving specificity of PI-PLC was examined by thin layer chromatography. Biochem J, 1989 May 1, 259(3), 913 - 6 Phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis as a probe for the distribution of phosphatidylinositol in hepatocyte membranes; Higgins JA et al.; Phosphatidylinositol-specific phospholipase C (PI-PLC) produced by Bacillus thuringiensis has been used as a probe for the distribution of phosphatidylinositol in hepatocyte membranes . Approx . 50% of this phospholipid was hydrolysed in microsomal vesicles (endoplasmic reticulum) with no significant hydrolysis of the remaining membrane phospholipids . Latency of mannose-6-phosphatase was retained during treatment indicating that the vesicles remained impermeable . Stripping of the ribosomes did not increase hydrolysis of phosphatidylinositol; however, when the vesicles were opened using dilute sodium carbonate, hydrolysis increased to greater than 90% . Hydrolysis of phosphatidylinositol of Golgi membranes was 35% and of plasma membranes was 50% . After treatment with PI-PLC, radiolabelled secretory proteins were retained in Golgi membranes and trapped lactate dehydrogenase was retained in plasma-membrane preparations indicating that the vesicles remained closed . Hydrolysis of phosphatidylinositol increased to greater than 90% when the membranes were opened by treatment with dilute sodium carbonate . These observations indicate that PI-PLC of Bacillus thuringiensis is a suitable probe for the distribution of phosphatidylinositol in membranes, and that in liver membranes this phospholipid occurs on each side of the bilayer, a topography consistent with its diverse roles. Biochim Biophys Acta, 1989 May 1, 995(3), 264 - 72 Production, purification and spectral properties of metal-dependent beta-lactamases of Bacillus cereus; Myers JL et al.; New methods for the production of consistently high levels of metal-dependent beta-lactamases (beta-lactamhydrolase, EC 3.5.2.6) from strains 569/H/9 and 5/B/6 of Bacillus cereus are described which have significant advantages over those reported previously . For example, these techniques do not require a fermentor with pH-stat capabilities . We also describe rapid very-high-yield purification schemes for the metal-dependent beta-lactamases from these strains, employing high-performance ultrafiltration (HPUF) and mass ion exchange techniques . Furthermore, we have developed improved methods for the removal of the active site Zn(II) and reconstitution of the beta-lactamase enzymatic activity with Co(II), which result in higher recovery of the original activity than previously reported . In order to characterize the purified beta-lactamases II of B . cereus 569/H/9 and 5/B/6 we have examined the molecular weights, and steady state kinetic parameters of Zn(II) enzymes, and the electronic and EPR spectra of the Co(II)-reconstituted enzymes . EPR spectra of CO(II)-reconstituted beta-lactamase from B . cereus 5/B/6 have not been previously reported. Curr Genet, 1989 May, 15(5), 363 - 70 Processing of chloroplast ribosomal RNA transcripts in Euglena gracilis bacillaris; McGarvey P et al.; The ribosomal RNA operons (rrn operons) of Euglena gracilis chloroplasts contain genes for (in order) 16S rRNA, tRNA(Ile), tRNA(Ala),23S rRNA and 5S rRNA . Major sites of cleavage of the primary rrn transcript were identified by Northern blot hybridization and S1-mapping . The presumptive termini of all of the mature products have now been identified . During initial processing in the chloroplast, the primary transcript is cleaved between the two tRNAs and between the 23S and 5S rRNAs so as to separate the sequences found in the different mature rRNAs . Subsequently the tRNAs are separated from the rRNAs, further trimming provides the remaining proper ends, and the 3'-ends of the tRNAs are added. Zentralbl Hyg Umweltmed, 1989 May, 188(1-2), 166 - 78 {Comparison of the sporocidal effects of two different disinfectants i a model experiment with pathogenic and nonpathogenic clostridial spore as well as with Bacillus cereus}; Stockinger H et al.; Official methods concerning the examination of sporocidal effects of disinfectants are presently lacking in the Federal Republic of Germany . Spore resistance of selected clostridial species towards formaldehyde and peracetic acid was examined by suspension tests and germ-carrier tests in comparison to spores of Bacillus cereus . Formaldehyde in concentrations between 5% and 20% as well as peracetic acid in concentrations between .004% and .125% were tested . Spore concentration of 10(6) spores/ml in the suspension-tests and 10(4) spores/ml in the germ-carrier tests with wooden germ-carriers were used . Against formaldehyde spores of Cl . septicum had the lowest resistance in the suspension tests followed by Cl . difficile, Cl . tertium, Cl . bifermentans and Cl . perfringens . Cl . sporogenes strain NCTC 532 had the most resistant spores . Concerning peracetic acid the situation was nearly the same . The resistance of all spores of pathogenic species under investigation towards chemical disinfectants was covered by those of Cl . sporogenes NCTC 532 treated with isopropanol . Isopropanol treated spores of B . cereus offer an alternative solution for practical disinfectant testing because spore production as well as growth of clostridia entail great expense . In order to cover the resistance of spores of Cl . perfringens A, the most resistant pathogenic clostridial species tested here, increased spore concentrations of B . cereus would be necessary. Protein Eng, 1989 May, 2(7), 559 - 62 Site-directed mutagenesis of glyceraldehyde-3-phosphate dehydrogenase reveals the role of residue Ser148; Corbier C et al.; A mutant of Bacillus stearothermophilus D-glyceraldehyde-3-phosphate dehydrogenase, Ser148----Ala, was produced by oligonucleotide-directed mutagenesis . The study of the catalytic properties of this mutant has shown that this mutation significantly affects the Michaelis constant of inorganic phosphate and to a lesser extent that of 1,3-diphosphoglycerate and D-glyceraldehyde-3-phosphate . This result is consistent with model-building studies which show that, for the phosphorylation step of catalysis, inorganic phosphate must bind to the anion recognition site designated Pi with the C(3) phosphate of the acyl-enzyme intermediate in the alternative anion site Ps . Studies of the enantiomeric specificity using D- and L-glyceraldehyde as substrates show that the hydroxyl group of Ser148, combined with the presence of the C(3) phosphate of the substrate, enhances stereospecificity as well as catalysis . However, the stereospecific effect cannot be a consequence of the direct interaction of Ser148 with the C(2)-hydroxyl of the substrate . The changed Km for glyceraldehyde-3-phosphate suggests that the initial step of hemithioacetal formation may take place with its C(3) phosphate bound in the Pi site . This supports the molecular mechanism proposed by Moody (1984) . Therefore, catalysis could be enhanced through interactions of the serine hydroxyl group not only with inorganic phosphate but also with the C(3) phosphate of glyceraldehyde-3-phosphate. Am J Clin Nutr, 1989 May, 49(5), 828 - 31 Viable starter culture, beta-galactosidase activity, and lactose in duodenum after yogurt ingestion in lactase-deficient humans; Pochart P et al.; Ten lactose malabsorbers were intubated and given fresh or heated yogurt to which polyethylene-glycol (PEG) and spores of Bacillus stearothermophilus (SBS) had been added as internal standards . In duodenal samples taken after fresh yogurt ingestion, viable starter culture was detected for 60 min in 6 of 7 subjects and the ratio of microbial beta-galactosidase activity to SBS remained similar during this period to its value in the preingested yogurt . In the two groups ingesting fresh and heated yogurt respectively, ratios of lactose to PEG remained similar to preingested values for 90 min and duodenal pH remained less than 5.1 . In vitro, at pH 5.0, beta-galactosidase activity in yogurt dropped by 80% . These data clearly show that after fresh yogurt ingestion, viable starter culture reaches the duodenum and contains beta-galactosidase activity . However, the buffering capacity of the yogurt that protects bacteria from acidic gastric secretion also prevents microbial beta-galactosidase from hydrolyzing lactose in the duodenum. Mikrobiologiia, 1989 May-Jun, 58(3), 365 - 9 {Isolation of mRNA from protoplasts of spore-forming bacteria and study of its translation products}; Sharipova FR et al.; Total RNA was isolated from the protoplasts of Bacillus intermedius 7P which actively produced alkaline exocellular RNAse . A fraction of polyadenylated RNA was isolated from this RNA using chromatography on poly(U) Sepharose . The total RNA and poly(A) +RNA were translated in Xenopus laevis oocytes . Bacterial exocellular RNAse was identified among the products of translation by means of electrophoretic and immunological analyses . The mRNA of RNAse secreted by B . intermedius 7P was concluded to be polyadenylated. Cancer, 1989 May 1, 63(9), 1676 - 80 Chemotherapy of disseminated melanoma with bleomycin, vincristine, CCNU, and DTIC (BOLD regimen) . The Prudente Foundation Melanoma Study Group. {Septic pneumonia caused by Bacillus cereus} Kovacs AM, Jozsef I. The authors describe the case of a 3-year old girl whose septic pneumonia was caused by B . cereus and developed following serial viral and bacterial infections, which produced assumably transient immunoppressed condition . Attention is drawn to the possibility of serious diseases caused by faculative bacteria in certain cases . The increasing pathological significance of bacteria existing in the soil is pointed out and the importance of hemoculture examination is stressed. Eur J Biochem, 1989 Apr 15, 181(1), 215 - 22 4-Chloroacetylpyridine adenine dinucleotide . A highly reactive and chromophoric affinity label of glyceraldehyde-3-phosphate dehydrogenase from sturgeon; Tritsch D et al.; The analogue of NAD+, 4-chloroacetylpyridine-adenine dinucleotide (clac4PdAD+), inactivated the glyceraldehyde-3-phosphate dehydrogenase from sturgeon at a high rate . An affinity labeling was shown to occur with clac4PdAD+ . The mononucleotide 4-chloroacetylpyridine 1-beta-D-ribose 5'-phosphate (clac4PdMN+) reacted with the enzyme in a second-order reaction whose rate was much smaller than that calculated for clac4PdAD+ taken as a second-order rate reagent . The rate of the reaction of clac4PdAD+ with the enzyme was determined by stopped flow, using as a probe the long-wavelength absorption maximum (430 nm) formed concomitantly with inactivation of the enzyme . Computer-assisted graphic simulation showed that the clac4PdAD+ analogue could bind to the active site of the enzyme from Bacillus stearothermophilus in a similar manner to that of NAD+, and that the reactive carbon and the reactive thiolate of Cys-149 were within bonding distance . The absorption at 430 nm was linearly proportional to the substoichiometric concentration of clac4PdAD+/mole subunit . Thiol titration suggested the modification of one thiol residue per subunit . The modified thiol was identified by degradation as Cys-149 . In contrast to the absorption band generated during the reaction of the 3-chloroacetylpyridine-adenine dinucleotide (clac3PdAD+) with the same enzyme {Eur . J . Biochem . (1982) 127, 519-524; 129, 437-446}, enzyme inactivated with clac4PdAD+ and clac4PdMN+ exhibited an absorption maximum at long wavelength which was still present after denaturation . The chromophore is proposed to be the enol form of the alpha-thioether ketone produced by alkylation of the thiolate of Cys-149 by the chloroacetyl group. J Biol Chem, 1989 Apr 15, 264(11), 6381 - 5 Stability-increasing mutants of glucose dehydrogenase from Bacillus megaterium IWG3; Makino Y et al.; A glucose dehydrogenase gene was isolated from Bacillus megaterium IWG3, and its nucleotide sequence was identified . The amino acid sequence of the enzyme deduced from the nucleotide sequence is very similar to the protein sequence of the enzyme from B . megaterium M1286 reported by Jany et al . (Jany, K.-D., Ulmer, W., Froschle, M., and Pfleiderer, G . (1984) FEBS Lett . 165, 6-10) . The isolated gene was mutagenized with hydrazine, formic acid, or sodium nitrite, and 12 clones (H35, H39, F18, F20, F191, F192, N1, N13, N14, N28, N71, and N72) containing mutant genes for thermostable glucose dehydrogenase were obtained . The nucleotide sequences of the 12 genes show that they include 8 kinds of mutants having the following amino acid substitutions: H35 and H39, Glu-96 to Gly; F18 and F191, Glu-96 to Ala; F20, Gln-252 to Leu; F192, Gln-252 to Leu and Ala-258 to Gly; N1, Glu-96 to Lys and Val-183 to Ile; N13 and N14, Glu-96 to Lys, Val-112 to Ala, Glu-133 to Lys, and Tyr-217 to His; N28, Glu-96 to Lys, Asp-108 to Asn, Pro-194 to Gln, and Glu-210 to Lys; and N71 and N72, Tyr-253 to Cys . These mutant enzymes have higher stability at 60 degrees C than the wild-type enzyme . The results of this study indicate that the tetrameric structure of glucose dehydrogenase is stabilized by several kinds of mutation, and at least one of the following amino acid substitutions stabilizes the enzyme: Glu-96 to Gly, Glu-96 to Ala, Gln-252 to Leu, and Tyr-253 to Cys. J Urol, 1989 Apr, 141(4), 853 - 6 Intravesical bacillus Calmette-Guerin therapy for in situ transitional cell carcinoma involving the prostatic urethra; Bretton PR et al.; A total of 23 patients presenting with multifocal superficial bladder cancer and concomitant in situ transitional cell carcinoma of the prostatic urethra (mucosal in 19 and ductal in 4) underwent transurethral resection and intravesical bacillus Calmette-Guerin therapy . Median followup was 51.6 months (range 6 to 105 months) . Of the 23 patients 13 (48 per cent) had a complete response with a median followup of 43.7 months without recurrence . Progression of some type (local, muscle invasion or metastasis) occurred in 10 patients (44 per cent); none occurred in the prostatic urethra . Median interval free of progression was 55.7 months; 7 of 10 patients required cystectomy for progression or refractory disease in the bladder (prostate negative for transitional cell carcinoma) . A trial of complete transurethral resection plus intravesical bacillus Calmette-Guerin is a viable alternative to immediate radical cystectomy for patients with mucosal and/or ductal involvement of the prostatic urethra with in situ transitional cell carcinoma. J Parasitol, 1989 Apr, 75(2), 191 - 4 Factors influencing lethality of Bacillus thuringiensis kurstaki toxin for eggs and larvae of Trichostrongylus colubriformis (Nematoda); Meadows J et al.; A toxin from Bacillus thuringiensis kurstaki was lethal to eggs and first- and second-stage larvae of the ruminant nematode Trichostrongylus colubriformis . Sheathed and exsheathed third-stage larvae were also killed by the toxin . However, susceptibility of the ova to the toxin decreased after several hours of development . Heating at 65 C for 1 hr or freezing at 0 C for 3 mo did not affect stability of the toxin . Ovicidal activity of the toxin was not altered by treatment with 13 microbial or mammalian enzymes, but toxicity was reduced by the antibiotics streptomycin or penicillin G and the enzyme inhibitor L-1-tosylamide 2-phenylethylchloromethyl ketone . Cuprous, ferrous, and zinc chlorides also inhibited the ovicidal activity of the toxin . Increased osmolarity of the assay media or solubilization of the toxin from pH 3 to 11 had no effect on toxicity for eggs . The membrane agents sodium vanadate and 4,4'-diisothiocyano-2,2' disulfonic acid stilbene increased (9-fold) and decreased (333-fold) toxicity, respectively . N-acetylneuraminic acid was the only tested sugar that reduced the toxicity of B . t . kurstaki. Mol Gen Mikrobiol Virusol, 1989 Apr, (4), 36 - 9 {Site-specific restriction endonuclease BtcI from Bacillus thuringiensis var . canadensis}; Kuzin AI et al.; Efficiency of bacteriophage Tp4 plating on Bacillus thuringiensis var . canadensis H5 (Can) is decreased 10(7)-fold as compared with the efficiency of plating on Bacillus thuringiensis var . galleriae H5 (Gal) . Bacteriophage Tp4 having propagated for one cycle in Can cells might be further grown in this strain without restriction . The sitespecific restriction endonuclease BtcI isolated from Bacillus thuringiensis var . canadensis recognises the same nucleotide sequence GATC in DNA as recognised by restriction endonuclease Sau3A. Pediatr Infect Dis J, 1989 Apr, 8(4), 210 - 5 Delayed cutaneous hypersensitivity: epidemiologic factors affecting and usefulness in predicting diarrheal incidence in young Peruvian children; Black RE et al.; Cell-mediated immunity, as assessed by delayed cutaneous hypersensitivity, can be diminished by malnutrition and viral infections . In turn, decreased immune functioning might lead to more frequent or more severe infectious diseases . Delayed cutaneous hypersensitivity was assessed in young Peruvian children by simultaneous application of seven standardized antigens and a negative control (Multitest CMI) . Response to tuberculin was frequent and was higher in children vaccinated with Bacillus Calmette-Guerin, response to tetanus or diphtheria toxoids was also good, especially in children who had received at least two doses of diphtheria-tetanus toxoids-pertussis vaccine . Two summary assessments, the number of positive responses and the sum of indurations of all positive responses provided useful measures of delayed cutaneous hypersensitivity . Responsiveness, as assessed by these summary measures, was inversely related to the incidence of diarrhea, identified by household surveillance for the 6 months after the skin test . Undernutrition, as assessed by weight for age or length for age, was also a significant determinant of the incidence of diarrhea, but not the duration of episodes, in this group of study children . Depressed cell-mediated immunity and malnutrition may be important risk factors for diarrhea in developing country children. Infect Control Hosp Epidemiol, 1989 Apr, 10(4), 150 - 4 Proteeae groin skin carriage in ambulatory geriatric outpatients; Ehrenkranz NJ et al.; Aerobic gram-negative bacillus (AGNB) groin skin carriage was prospectively studied in ambulatory geriatric outpatients: 42 from three nursing homes and 44 from private homes . Initially, 12 (28.6%) Proteeae carriers were in the former group and 3 (6.8%) were in the latter (P = 0.01) . At one year, 6 of 7 surviving nursing home carriers remained Proteeae carriers while none from private homes remained carriers (P = 0.007) . The annual prevalence of Proteeae carriage was 14 (33.3%) in nursing homes and 4 (9.1%) in private homes (P = 0.008); of non-Proteeae AGNB carriage, the annuyal prevalence was 2 (4.8%) and 4 (11.4%), respectively . Nursing home subjects had similar initial health characteristics; however, by one year, 5 of 12 carriers in contrast to 3 of 30 noncarriers were dead of chronic disease (P = 0.03) . These nursing homes included persons with chronic diseases that apparently facilitated Proteeae carriage . Urethral catheters, skin ulcers, and recent antibiotics were not factors. J Biochem (Tokyo), 1989 Apr, 105(4), 653 - 9 Reaction mechanism of gramicidin S synthetase 1, phenylalanine racemase, of Bacillus brevis; Kanda M et al.; We have demonstrated that gramicidin S synthetase 1 (GS 1), phenylalanine racemase {EC 5.1.1.11}, of Bacillus brevis catalyzes the exchange between a proton in the medium and alpha-hydrogen of phenylalanine in the course of the racemase reaction by using tritiated water or L-phenyl{2,3-3H}alanine . GS 1 from some gramicidin S non-producing mutants of B . brevis lacking phenylalanine racemase activity did not catalyze the tritium exchange reaction . The proton exchange between phenylalanine bound as thioester on the GS 1-phenylalanine complex and water in the medium was detected, but 5,5'-dithiobis(2-nitrobenzoic acid)-modified complex lacked both the proton exchange and phenylalanine racemase activity . It is suggested that a base group, probably a sulfhydryl group, on the enzyme functions as proton donor and acceptor during the phenylalanine racemase reaction. J Biochem (Tokyo), 1989 Apr, 105(4), 626 - 32 Tryptic digestion of NADH dehydrogenase from alkalophilic Bacillus; Xu XM et al.; The alkalophile NADH dehydrogenase (NADH: 2,6-dichlorophenolindophenol oxidoreductase) {EC 1.6.99.3} consists of two identical subunits of 65 kDa, and each subunit contains the catalytic and liposome-binding regions . On treatment with trypsin, the polypeptide exhibiting the liposome-binding property in one of the subunits was digested to form an enzymatically active hetero-dimer (40 and 65 kDa), and then the polypeptide in the other subunit was digested to form an active homo-dimer (40 and 40 kDa) . The hetero-dimer bound to liposomes, but the homo-dimer did not . Kinetic analysis showed that removal of one or two of the polypeptides in the enzyme slightly affects its kinetic parameters . For all the enzyme species, NAD inhibited competitively with respect to NADH and non-competitively with respect to 2,6-dichlorophenolindophenol . The partially determined amino acid sequence of this alkalophile enzyme suggested that (i) a long random-coiled peptide (58 amino acid residues) or a portion of the peptide is located between the polypeptides with liposome-binding and catalytic properties, (ii) the polypeptide exhibiting liposome-binding property is in the amino terminal region of the enzyme, (iii) the amino acid sequences around the subtilisin and trypsin cleavage sites of the peptide are hydrophilic and on the surface of the protein molecule and therefore are susceptible to digestion, and (iv) the FAD-binding site is located near the amino terminal region of the catalytic region. Antibiot Khimioter, 1989 Apr, 34(4), 266 - 70 {In vitro and in vivo studies of RNAse of Bacillus intermedius}; Kurinenko BM et al.; Cytotoxicity of RNAase from Bacillus intermedius was studied in vitro and in vivo . It was shown that the enzyme had slightly pronounced cytotoxicity according to the tests with inhibition of cell proliferation and biosynthesis of cell nucleic acids . The RNAase was also shown to impair the vital staining by neutral red . The efficiency of the impairment much more depended on the enzyme catalytic activity than on the proliferation and biosynthesis of nucleic acids . In vivo toxicity of RNAase from B . intermedius was 3-5 times higher than that of pancreatic RNAase . Possible mechanisms of the different toxicity of the enzymes are discussed. Indian J Lepr, 1989 Apr, 61(2), 229 - 32 Status of multi bacillary patients "lost to follow up"; Muliyil J et al.; Completeness of coverage with MDT is essential if one were to hope for a reduction in the incidence of leprosy in an endemic area . Patients who are lost to follow up are generally declared as "Permanently left" (PL) and deleted from the known case register . A special effort was made to study the true status of multibacillary patients who had been declared "PL" in the leprosy control programme area of the Christian Medical College, Vellore . 38% of the 40 patients followed up were found to be still residing in the same area . 40% of them were BI Positive . The reasons for these erroneous deletions varied . The study shows that declaration of "PL" need to be verified carefully . Further, it was found that the present system of reporting may exaggerate the actual number of patients who have been lost to follow up. FEMS Microbiol Lett, 1989 Apr, 49(2-3), 157 - 63 Delineation of the toxin coding fragments and an insect-specificity region of a dual toxicity Bacillus thuringiensis crystal protein gene; Haider MZ et al.; A series of deletion mutants have been constructed from the dual toxicity Bacillus thuringiensis aizawai IC1 (Bta IC1) crystal protein gene . The mutant toxin genes were expressed in Escherichia coli, their protein products purified and the authenticity of these mutant proteins confirmed immunologically . Analysis of the toxicity spectra of these mutants revealed that lepidopteran toxicity is located on the N-terminal region of the toxin between residues Ile30-Glu595 . 3' deletion of a further 37 residues from Glu595 of the lepidopteran-specific toxin abolished lepidopteran toxicity but the resulting protein consisting of residues Ile30-Gly558 was still fully toxic to dipteran larvae and cells . Another mutant crystal protein gene truncated to encode residues between Ile30-Gly563 was toxic only to diptera . These data indicate that the determinants of lepidopteran specificity in the Bta IC1 toxin are located between residues Gly558-Glu595 and that the N-terminal portion of the toxin between Ile30-Gly558 is sufficient to express dipteran toxicity. FEMS Microbiol Lett, 1989 Apr, 49(2-3), 121 - 4 A screening for the presence of four different crystal protein gene types in 25 Bacillus thuringiensis strains; Visser B; 25 different strains of Bacillus thuringiensis, belonging to 18 different serotypes were screened by Southern hybridization analysis for the presence of nucleotide sequences of four different gene types coding for crystal proteins showing different insecticidal spectra . Many but not all strains showed sequences homologous to any of the probes used . Homology with sequences of the gene type present in strain kurstaki HD1 occurred in all positive strains, whereas the other gene types were much less abundant . Sequences homologous to those of gene type VI, coding for a Spodoptera-specific crystal protein, appeared only in strains of serotypes aizawai and entomocidus. Can J Microbiol, 1989 Apr, 35(4), 444 - 9 {Biochemical diagnosis of the presence of poisoning by Bacillus thuringiensis serotype "H3a,3b" in two lepidopterans}; Valero JR et al.; A detailed biochemical analysis has shown that during larval development on artificial medium, the amounts of K+, Na+, and Ca2+ in the hemolymph of healthy Choristoneura fumiferana varied from 85 to 110 mg/100 mL, 29 to 33 mg/100 mL, and 4.8 to 7.3 mg/100 mL, respectively . Similar results were obtained with Malacosoma disstria . Intoxication by Bacillus thuringiensis "H3a,3b" (B . t.) considerably modified the amounts of the cations . Thus, after 4 h, the quantity of K+ in M . disstria increased from 99 to 229 mg/100 mL and Na+ from 26.5 to 50.3 mg/100 mL while that of Ca2+ decreased from 5.8 to 1.2 mg/100 mL . Similar results were observed with C . fumiferana, but these variations occurred after 2 to 4 days of B . t . intoxication . The variations detected during the bacillosis, with respect to the cationic composition of the insect hemolymph, are rapidly detectable, well before light microscope observation can confirm the presence of this intoxication . Aspartate aminotransferase, alanine aminotransferase, alpha-hydroxybutyrate dehydrogenase, and isocitrate dehydrogenase activity fluctuated very slightly in the hemolymph of either healthy or bacillosed larvae of the two insects under study . These results suggest that it is possible to diagnose biochemically the presence of a B . t . intoxication in lepidopteran forest pests following treatments by this biological insecticide for their control. Vaccine, 1989 Apr, 7(2), 97 - 101 Anti-fertility vaccines; Talwar GP et al.; Vaccines are under development for the control of fertility in males and females . This review discusses developments in anti-fertility vaccines at the National Institute of Immunology, New Delhi, India . A single injection procedure for the sterilization or castration of male animals depending on the site at which the injection is given, has passed through field testing and is expected to be on the market in the near future . Vaccines inducing antibodies against the human chorionic gonadotropin have gone through phase I trials with satisfactory results . A vaccine producing a consistently bioeffective antibody response against gonadotropin-releasing hormone is ready for phase I/II clinical trials in patients of carcinoma of prostate after due experimentation in animals and toxicology studies . Research to identify sperm antigens for incorporation into second generation vaccines is in progressPIP: Anti-fertility vaccine research carried out at the National Institute of Immunology in New Delhi, India is summarized . The vaccines react against follicle stimulating hormone (FSH), luteinizing hormone (LH), and gonadotropin releasing hormone (GnRH) . Intratesticular injection of Bacillus Calmette-Guerin (BCG) in moderate concentrations causes the degeneration of the blood-testes barrier, which finally leads to reversible azoospermia and it is suited for use in animals . Anti-hCG (anti-human chorionic gonadotropin) vaccines include beta-hCG linked to tetanus toxoid /TT (beta-hCG-TT) the alpha-subunit of ovine LH associated with beta-hCG and linked to TT or cholera toxin chain b (CHB/alpha-oLH, beta-hCG-TT/CHB/, the mixture of hCG and oLH, and 37 amino acid carboxy-terminal peptide (CTP) of beta-hCG linked to diphtheria toxoid (DT) . The principle of anti-hCG vaccine is to induce antibodies which can bind to hCG and render it biologically inactive . The beta-subunit of hCG has proven efficacious in controlling fertility, and similarly 36 amino acid CTP was effective in baboons . The beta-hCG- TT vaccine evoked the production of anti-hCG antibodies in 61 of 63 women tested . The mixture of beta-hCG and beta-ovine LH conjugated to TT or CHB /formulation M/ and beta-hCG annealed to alpha-ovine LH and conjugated to TT or to CHB (formulation A/ produced improved) immunogenicity . Immunization with GnRH resulted in decreased testicular size, drop of testosterone levels, and marked atrophy of the prostate . Anti-GnRH vaccine would be tried in patients of carcinoma of the prostate . LDH-C4, a sperm-specific mitochondrial antigen, produced an antibody response in baboons and reduced fertility in the females . Animal fertility control vaccines will be shortly on the market, but the use of recombinant DNA techniques should also accelerate the development of others . Pediatr Infect Dis J, 1989 Apr, 8(4), 201 - 6 Clinical presentation of Bacillus Calmette-Guérin infections in patients with immunodeficiency syndromes; Gonzalez B et al.; Nine children with immunodeficiency syndromes who developed persistent or disseminated Bacillus Calmette-Guerin (BCG) infections after BCG vaccination at birth were observed in Santiago, Chile, over a period of 10 years . This represents a risk for persistent or disseminated BCG infections of 3.4/1,000,000 vaccinated newborns . This may closely reflect the incidence of severe combined immunodeficiency syndromes, cellular immunodeficiency syndromes and chronic granulomatous disease in the study area . The clinical presentation and course of the infection varied considerably depending on the underlying immunodeficiency syndrome . Two patients with severe combined immunodeficiency presented with cutaneous nodules in the absence of any local reaction at the site of BCG vaccination . Both patients died of disseminated BCG infection within the first year of life . Four patients with cellular immunodeficiency syndromes presented with regional lymphadenitis resistant to treatment after the fifth month of life . Three of these patients had specific unresponsiveness to tuberculin and survived from 5 to 6 years of age . Two boys with X-linked chronic granulomatous disease presented with regional lymphadenitis in the first 3 months of life . A girl with autosomal recessive chronic granulomatous disease presented at 18 months of age with regional lymphadenitis . All three patients with chronic granulomatous disease had positive tuberculin reactions and died from infections other than BCG. FEMS Microbiol Lett, 1989 Apr, 49(2-3), 171 - 7 Transformation of Bacillus thuringiensis by electroporation; Bone EJ et al.; Plasmids were transformed by electroporation into various strains of Bacillus thuringiensis with frequencies of up to 10(5) transformants/micrograms . pC 194 transformed all strains tested at a high frequency and cells could be stably transformed with pC194 and pUB110 simultaneously by electroporation with a frequency of 10(2) pC194+ pUB110 transformants/micrograms DNA . Low transformation frequencies observed with some plasmids, especially those grown initially in Escherichia coli, could be increased by passage through B . thuringiensis, B . thuringiensis var . israelensis and in acrystalliferous mutant of the same strain transformed at frequencies of 10(4)-10(5)/micrograms DNA with most of the plasmids tested . A cloned israelensis 27-kDa delta-endotoxin gene was introduced into the israelensis acrystalliferous mutant and a kurstaki acrystalliferous mutant by electroporation . Both transformants were shown to express the endotoxin gene and to be toxic to Aedes aegypti larvae. Antonie Van Leeuwenhoek, 1989 Apr, 55(4), 303 - 11 Purification and characterization of the NADP-dependent glutamate dehydrogenase from Bacillus fastidiosus; Op den Camp HJ et al.; Ammonia assimilation in Bacillus fastidiosus proceeds via the NADP-dependent glutamate dehydrogenase . The enzyme, purified to homogeneity, is composed of identical subunits with a molecular weight of about 48,000 dalton . Presumably the enzyme is a hexamer . The enzyme is specific for NADP(H) . The pH optima for the amination and deamination reactions are 7.7 and 8.6, respectively . The temperature optimum is 60 degrees C . Furthermore, temperature stability and apparent Km values for substrates of both the amination and deamination reactions were determined . Several metabolites were tested for their effect on the enzyme activity . Only malate and fumarate showed some inhibitory effect. J Bacteriol, 1989 Apr, 171(4), 1879 - 84 The sodium/proton antiport system in a newly isolated alkalophilic Bacillus sp; Kitada M et al.; The pH homeostasis and the sodium/proton antiport system have been studied in the newly isolated alkalophilic Bacillus sp . strain N-6, which could grow on media in a pH range from 7 to 10, and in its nonalkalophilic mutant . After a quick shift in external pH from 8 to 10 by the addition of Na2CO3, the delta pH (inside acid) in the cells of strain N-6 was immediately established, and the pH homeostatic state was maintained for more than 20 min in an alkaline environment . However, under the same conditions, the pH homeostasis was not observed in the cells of nonalkalophilic mutant, and the cytoplasmic pH immediately rose to pH 10 . On the other hand, the results of the rapid acidification from pH 9 to 7 showed that the internal pH was maintained as more basic than the external pH in a neutral medium in both strains . The Na+/H+ antiport system has been characterized by either the effect of Na+ on delta pH formation or 22Na+ efflux in Na+-loaded right-side-out membrane vesicles of strain N-6 . Na+- or Li+-loaded vesicles exhibited a reversed delta pH (inside acid) after the addition of electron donors (ascorbate plus tetramethyl-p-phenylenediamine) at both pH 7 and 9, whereas choline-loaded vesicles generated delta pHs of the conventional orientation (inside alkaline) . 22Na+ was actively extruded from 22Na+-loaded vesicles whose potential was negative at pH 7 and 9 . The inclusion of carbonyl cyanide m-chlorophenylhydrazone inhibited 22Na+ efflux in the presence of electron donors . These results indicate that the Na+/H+ antiport system in this strain operates electrogenically over a range of external pHs from 7 to 10 and plays a role in pH homeostasis at the alkaline pH range . The pH homeostasis at neutral ph was studied in more detail . K+ -depleted cells showed no delta pH (acid out) in the neutral conditions in the absence of K+, whereas these cells generated a delta pH if K+ was present in the medium . This increase of internal pH was accompanied by K+ uptake from the medium . These results suggest that electrogenic K+ entry allows extrusion of H+ from cells by the primary proton pump at neutral pH. Mol Microbiol, 1989 Apr, 3(4), 487 - 95 The high-affinity K+-translocating ATPase complex from Bacillus acidocaldarius consists of three subunits; Hafer J et al.; Cells of the thermoacidophilic bacterium Bacillus acidocaldarius express a high-affinity K+-uptake system when grown at low external K+ . A vanadate-sensitive, K+- and Mg2+-stimulated ATPase was partially purified from membranes of these cells by solubilization with a non-ionic detergent followed by ion-exchange chromatography of the extract . Combinations of non-denaturing and denaturing electrophoretic separation methods revealed that the ATPase complex consisted of three subunits with molecular weights almost identical to those of the KdpA, B and C proteins, which together form the Kdp high-affinity, K+-translocating ATPase complex of Escherichia coli . The affinity of the partially purified ATPase from B . acidocaldarius for its substrates K+ (Km 2-3 microM) and ATP (Km 80 microM), its stimulation by various divalent cations, and its inhibition by vanadate (Ki 1-2 microM), bafilomycin A1 (Ki 20 microM), DCCD (Ki 200 microM) or Ca2+ were also similar to those of the E . coli enzyme, indicating that the two K+-translocating ATPases have almost identical properties. Lab Anim, 1989 Apr, 23(2), 96 - 102 Transmission experiments of cilia-associated respiratory bacillus in mice, rabbits and guineapigs; Matsushita S et al.; Transmission experiments of cilia-associated respiratory (CAR) bacillus were performed in mice in order to clarify the principal route of the infection, and in rabbits and guineapigs in order to examine their susceptibility . Determination of the infection was evaluated serologically by the indirect immunofluorescence assay (IFA) technique and histologically by the presence of CAR bacillus in the airways . BALB/c mice were intranasally inoculated with the SMR strain of CAR bacillus . The IFA antibody to the bacteria in these mice rose to more than 1:160 at 4 weeks postinoculation (PI) and the mice were utilized as transmitters for the following experiments . One out of 15 uninfected mice kept in intracage contact with infected mice became infected from 4 weeks after contact . Incidence of contact infection increased thereafter . On the other hand, there was no evidence of infection in the uninfected mice housed in the separate cages from the cage in which infected mice were housed throughout the 12-week observation period . The primary method of CAR bacillus transmission seems to be direct contact with infected mice or fomites contaminated by infected mice; airborne transmission appears to be of little importance . Rabbits and guineapigs were also intranasally inoculated with the SMR strain of CAR bacillus . IFA antibodies were positively detected by 4 weeks PI, but no CAR bacillus nor histological changes relating to the infection were observed in the airways of either species . It is suggested that rat origin CAR bacillus can transmit to rabbits and guineapigs, and that the infection can spread to other species of rodents and rabbits. FEBS Lett, 1989 Mar 27, 246(1-2), 83 - 8 Interaction of BlaI, the repressor for the beta-lactamase gene of Bacillus licheniformis, with the blaP and blaI promoters; Grossman MJ et al.; BlaI repressor for the beta-lactamase gene (blaP) of Bacillus licheniformis 749, was shown to repress expression of blaP and of the repressor gene (blaI), using the purified protein in a DNA-directed in vitro translation assay . Binding of BlaI to the promoter regions of blaP and blaI was examined by equilibrium and competitive binding assays . BlaI binds to the blaP promoter with an equal or only slightly higher affinity than to the blaI promoter . DNase I footprinting shows that BlaI binds directly to the blaP and blaI promoters, such that RNA polymerase binding and/or transcript elongation would be blocked. Nature, 1989 Mar 23, 338(6213), 357 - 60 High-resolution (1.5 A) crystal structure of phospholipase C from Bacillus cereus; Hough E et al.; Both the phosphatidylinositol-hydrolysing and the phosphatidylcholine-hydrolysing phospholipases C have been implicated in the generation of second messengers in mammalian cells . The phosphatidylcholine-hydrolysing phospholipase C (PLC) from Bacillus cereus, a monomeric protein containing 245 amino-acid residues, is similar to some of the corresponding mammalian proteins . This, together with the fact that the bacterial enzyme can mimic the action of mammalian PLC in causing, for example, enhanced prostaglandin biosynthesis, suggests that B . cereus PLC can be used as a model for the hitherto poorly characterized mammalian PLCs . We report here the three-dimensional structure of B . cereus PLC at 1.5 A resolution . The enzyme is an all-helix protein belonging to a novel structural class and contains, at least in the crystalline state, three Zn2+ in the active site . We also present preliminary results from a study at 1.9 A resolution of the complex between PLC and inorganic phosphate (Pi) which indicate that the substrate binds directly to the metal ions. Lancet, 1989 Mar 18, 1(8638), 601 - 3 Primary cutaneous bacillus cereus infection in neutropenic children; Henrickson KJ et al.; A review of culture reports for a five-year period at St Jude Children's Research Hospital yielded 10 cases of primary cutaneous Bacillus cereus infection in neutropenic patients treated for cancer or aplastic anaemia . Vesicles or pustules were seen only on the limbs . The infections, all of which arose in the spring or summer, responded to antibiotics . In neutropenic patients B cereus should thus be regarded as a possible cause of isolated vesicles. Gene, 1989 Mar 15, 76(1), 137 - 44 Multiple amylase genes in two strains of Bacillus stearothermophilus; Sen S et al.; Plasmid DNA fragments from Bacillus stearothermophilus ATCC29609 (BR135) and chromosomal DNA fragments from B . stearothermophilus ATCC31195 (BR132) were cloned into pBR322 and transformed into Escherichia coli strain HB101 . Clones were selected which demonstrate extracellular expression of thermostable amylase . The DNA inserts from both strains showed similar restriction maps . Examination of the parental strains revealed plasmids of approx . 100 kb and 35 kb for BR135 and no discernible plasmids for BR132 . Hybridization experiments showed that the amylase gene is contained within a 3.2-kb HindIII fragment on both plasmids, and to be present in multiple copies in the BR135 chromosome . Multiple chromosomal copies of the amylase gene were also observed for BR132 . Purification of the alpha-amylase produced by the cloned plasmid amy gene yielded a homogeneous 58-kDa protein with thermostable amylase activity. Anal Chem, 1989 Mar 15, 61(6), 624 - 7 Evaluation of the metal uptake of several algae strains in a multicomponent matrix utilizing inductively coupled plasma emission spectrometry; Mahan CA et al.; Three freshwater heat-killed, lyophilized blue-green algae strains have been characterized as to their ability to accumulate heavy metals with a focus on the utilization of these algae as an analytical preconcentration technique . This study examines the metal uptake in several multicomponent mixtures by using inductively coupled plasma optical emission spectrometry (ICP-OES) . Six milligrams of a pure strain of algae was added to 20-mL aliquots of buffered (pH 5.5-6.5) multielement solutions containing 0.1, 0.5, 1.0, 2.0, and 4.0 mg/L of K, Mg, Ca, Fe, Sr, Co, Cu, Mn, Ni, V, Zn, As, Cd, Mo, Pb, and Se . All three algae strains exhibit relatively high adsorption affinities for Fe, Pb, and Cu, with uptake between 70 and 98% at the 4 ppm concentration level . Biosorption occurs for essentially every element with the relative affinities decreasing in the order Pb greater than Fe greater than Cu greater than Cd greater than Zn greater than Mn greater than Mo greater than Sr greater than Ni greater than V greater than Se greater than As greater than Co for Chlorella pyrenoidosa at the 4 mg/L concentration level . Although some minor differences were seen, the other algae strains (Stichococcus bacillaris and Chlamydomonas reinharti) displayed similar adsorption behavior over the concentration range studied, indicating similar cell wall binding sites . Langmuirian isotherms exhibited a minimum of two slopes over the concentration range of 0.1-4.0 mg/L, indicating the probable existence of at least two adsorption mechanisms. Biochem J, 1989 Mar 15, 258(3), 715 - 21 Probing the substrate-binding sites of aminoacyl-tRNA synthetases with the procion dye green HE-4BD; McArdell JE et al.; A reactive bis-dichloro derivative of the Procion dye Green HE-4BD was shown to inactivate irreversibly methionyl-tRNA synthetase (MTS) from Escherichia coli and also tryptophyl-tRNA synthetase (WTS) and tyrosyl-tRNA synthetase (YTS) from Bacillus stearothermophilus at pH 8.5 and 37 degrees C . At a 5-fold excess of reactive dye over enzyme subunit concentration MTS was quantitatively inactivated within 20 min in the ATP/pyrophosphate exchange assay, whereas WTS and YTS show an 80% loss of activity over the same time period . The inactivation is affected by the addition of substrates, which either protect (WTS and YTS) or promote (YTS with tyrosine) the dye-mediated enzyme inactivation . Green HE-4BD-OH was shown to be a competitive inhibitor of MTS with respect to MgATP, methionine and tRNA substrates. Eur J Biochem, 1989 Mar 15, 180(2), 309 - 18 Oxacillin-hydrolysing beta-lactamases . A comparative analysis at nucleotide and amino acid sequence levels; Mossakowska D et al.; We have extended the sequence of the OXA-2 beta-lactamase which together with S1 mapping has enabled us to identify the promoter site for this gene . This lies in a region that is found upstream from a variety of resistance genes on different plasmids; each gene appears to have been inserted at the same specific site and to be expressed from the same promoter . The ancestral plasmid thus appears to function as a natural expression vector . The sequence of the recombination site at the 5' end of the OXA-2 gene shows a marked similarity with the attP sequence of lambda . DNA-probe analysis confirmed that the OXA-2 and OXA-3 beta-lactamases are related, and indicated no similarity with other beta-lactamase genes . However, a comparison of amino acid sequences demonstrates that the OXA-2, OXA-1 and PSE-2 beta-lactamases show some similarities to the typical class A enzymes, especially in the central helical domain of the latter, which is largely responsible for forming the active site of the enzyme . The three oxacillinases also show marked amino acid sequence similarity with the product of a regulatory gene, blaR1, required for beta-lactamase induction in Bacillus licheniformis. Biochem J, 1989 Mar 15, 258(3), 765 - 8 A thiono-beta-lactam substrate for the beta-lactamase II of Bacillus cereus . Evidence for direct interaction between the essential metal ion and substrate; Murphy BP et al.; An 8-thionocephalosporin was shown to be a substrate of the beta-lactamase II of Bacillus cereus, a zinc metalloenzyme . Although it is a poorer substrate, as judged by the Kcat./Km parameter, than the corresponding 8-oxocephalosporin, the discrimination against sulphur decreased when the bivalent metal ion in the enzyme active site was varied in the order Mn2+ (the manganese enzyme catalysed the hydrolysis of the oxo compound but not that of the thiono compound), Zn2+, Co2+ and Cd2+ . This result is taken as evidence for kinetically significant direct contact between the active-site metal ion of beta-lactamase II and the beta-lactam carbonyl heteroatom . No evidence was obtained, however, for accumulation of an intermediate with such co-ordination present. Indian J Malariol, 1989 Mar, 26(1), 25 - 31 Evaluation of Bacillus sphaericus to control breeding of malaria vectors; Ansari MA et al.; Bacillus sphaericus formulations (Solvay liquid 2362 and Abbott granules 2297) were tested in the laboratory and field for the control of mosquito breeding . Results of laboratory evaluation revealed that both formulations had good larvicidal activity against Culex quinquefasciatus, at 0.04 ml and 1 gm/sq m dosages respectively . Higher dosages at 5 ml liquid or 5 gm/sq m granules were required to control A . stephensi and A . subpictus . A . culicifacies required still higher dosages of Solvay (25 ml/sq m) to achieve same levels of control whereas Abbott granules were not effective even at 20 gm/sq m. Pneumologie, 1989 Mar, 43(3), 169 - 72 {Lung diseases caused by non-tuberculous mycobacteria in East Germany and Hungary}; Kalich R et al.; Despite the decrease of new cases of pulmonary tuberculosis also between 1982 and 1986 an increase in nontuberculous mycobacteriosis can be seen especially in the GDR and also in Hungary . In 1986 the ratio of recognized lung diseases caused by nontuberculous mycobacteria to all newly diagnosed bacillary cases was 4.1% in the GDR and 2.2% in Hungary . In Hungary, M . xenopi was the most important potential pathogenic species of nontuberculous mycobacteria during the whole period, followed by M . kansasii . In the GDR at the beginning most cases of nontuberculous mycobacteriosis were caused by M . kansasii later, M . xenopi was most frequent . Most of the patients with nontuberculous mycobacteriosis were men older than 50 years of age. Ann Soc Belg Med Trop, 1989 Mar, 69(1), 67 - 73 {{Potential and limitations of the long-term treatment of bacilliferous tuberculosis in Maniema (Zaire)}; De Caluwe P et al.; Results after one year treatment of tuberculosis with a regimen associating streptomycin, isoniazid and thiacetazone in a rural region of eastern Zaire are analyzed . A 46.9% cure rate is found . A high death rate (21.5%) and a relatively low number of "lost" patients (13.8%) are noted . The usefulness of the introduction of short term chemotherapy is discussed. Mol Gen Mikrobiol Virusol, 1989 Mar, (3), 25 - 8 {Intraspecies plasmid transformation of Bacillus thuringiensis}; Bessol'tsev VV et al.; Bacillus thuringiensis strains 1009, 3004, 3029 of the natural isolates of var . dendrolimus harbouring two large plasmids were transformed by a mix of small plasmids DNA . One of transforming plasmids carried a gene for tetracyclinresistance . The natural isolates of var . dendrolimus strain 2002 and var . berliner strain 1035 were used as donors for plasmid DNA transformation . Three variants of plasmid spectra registered in tetracycliresistant transformants are discussed. J Invertebr Pathol, 1989 Mar, 53(2), 206 - 16 Thuricin: the bacteriocin produced by Bacillus thuringiensis; Favret ME et al.; Bacillus thuringiensis serovar, thuringiensis (HD-2) demonstrated antibacterial activity against 48 of 56 strains of B . thuringiensis and against some other Gram-positive species but not against Gram-negative species . The antibacterial activity was not inducible by mitomycin C or by ultraviolet irradiation, and additional activity was not liberated from cells by sonication . Upon dilution of the antibacterial substance, zones of inhibition diminished without the appearance of plaques . Gel filtration chromatography indicated an Mr greater than 950,000 for the bacteriocin (thuricin) in its native form . The native thuricin was sedimented by ultracentrifugation, but electron microscopy of the pellet failed to reveal phage particles or phage components . Nondenaturing polyacrylamide gel electrophoresis (PAGE) of thuricin demonstrated the association of bacteriocin activity with a protein band which migrated only slightly into a 5% gel . Sodium dodecyl sulfate (SDS)-PAGE of partially purified thuricin revealed five major bands . Thuricin activity was substantially reduced by treatment with chymotrypsin, pronase, subtilisin, trypsin, and heat at 96 degrees C but not by treatment with lysozyme, phospholipase C, papain, peptidase, or organic solvents . It exhibited a bactericidal and bacteriolytic effect on a sensitive strain, B . thuringiensis serovar, canadensis (MF4) . Partially purified preparations of thuricin had phospholipase A activity which was adsorbed by sensitive cells but not by cells which were insensitive to thuricin . Antibacterial activity was blocked by preincubation of thuricin with phospholipid . Loss of a 150-mDa plasmid was correlated with loss of thuricin production. FEMS Microbiol Lett, 1989 Mar, 49(1), 1 - 4 Low heat resistance of Bacillus sphaericus spores correlated with high protoplast water content; Beaman TC et al.; The low heat resistance (D100 = 0.554 min, z = 13.4 degrees C) of dormant lysozyme-sensitized spores of Bacillus sphaericus 9602 was correlated with a low protoplast wet density (1.305 g/ml) equivalent to a high protoplast water content (61.0%, wet weight basis) . These values for these unusual spores were consistent with those correlated previously in 28 spore types of seven other species. Rev Infect Dis, 1989 Mar-Apr, 11(2), 349 - 51 Does vaccination with bacille Calmette-Guérin protect against AIDS? Kallenius G, Hoffner SE, Svenson SB. In the United States a majority of patients with AIDS are infected with bacteria of the Mycobacterium avium complex (MAC), while in Sweden only approximately 10% of AIDS patients are so infected . It is proposed that general vaccination with bacille Calmette-Guerin (BCG) in Sweden may have induced protection not only against tuberculosis but also against infections with MAC, accounting for the lower incidence of MAC infection in Swedish patients with AIDS . The current AIDS pandemic may indicate a need for reevaluation of national BCG vaccination policies. Bull Int Union Tuberc Lung Dis, 1989 Mar, 64(1), 8 - 11 Tuberculosis and AIDS . Statement on AIDS and tuberculosis . Geneva, March 1989 . Global Programme on AIDS and Tuberculosis Programme, World Health Organization, in collaboration with the International Union Against Tuberculosis and Lung Disease; Interactions of the human immunodeficiency virus and tuberculosis and the implications for BCG vaccination; Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, MarylandAIDS has become a global epidemic, with greater than 100,000 cases officially reported in 140 countries and an estimated 5-10 million asymptomatic carriers of the human immunodeficiency virus (HIV), the etiologic agent of AIDS . With an increase in HIV infection in some developing countries, there has been a resurgence in tuberculosis, and concern has been raised about the indications, efficacy, and safety of bacille Calmette-Guerin (BCG) vaccination . Anecdotal reports of local reactions and disseminated disease have been described in HIV-infected children and adults . Ten HIV-infected infants, who received BCG vaccination within 2 months of birth, developed local lymphadenitis at 4-15 months . However, in one preliminary survey in Zaire, the rates of local lymphadenitis were equal in HIV-infected and HIV-uninfected children, and no dissemination has been observed to date . Until further information is known, BCG vaccinations should not be given to symptomatic HIV-infected individuals and should only be given to HIV-infected children who are asymptomatic and who reside in areas where tuberculosis is highly endemic and where the risk of tuberculosis may outweigh the potential complications of BCG immunization. Rev Infect Dis, 1989 Mar-Apr, 11 Suppl 2, S360 - 5 Impact of present control methods on the problem of tuberculosis; Pio A; In spite of the considerable efforts that have been made to control tuberculosis in developing countries in the last three decades, the decline in the global epidemiologic problem has been slow . It is now realized that the possibility of rapidly influencing the tuberculosis problem in these countries with the present control technologies has been overestimated . It is clear that bacille Calmette-Guerin (BCG) vaccination, even in places where a high efficacy was demonstrated, does not substantially influence the chain of transmission . Case management, including bacteriologic diagnosis and chemotherapy, is a complex technology . Although it is potentially effective for the reduction of mortality and the risk of infection, there are many difficulties for its efficient application in developing countries . Although there should be no hesitation in the application and full utilization of what is already known, a major thrust in research may generate ways of accelerating the control of tuberculosis in developing countries . Without new developments in technology for tuberculosis control, the goal of worldwide control has still a long way to go. Rev Infect Dis, 1989 Mar-Apr, 11 Suppl 2, S353 - 9 The BCG story: lessons from the past and implications for the future; Fine PE; BCG (bacille Calmette-Guerin) vaccines are at once among the least satisfactory and yet the most widely used of all vaccines today . Their variable efficacy against tuberculosis and leprosy is still not understood and points to a fundamental unsolved problem in vaccine immunology . The extensive use of BCG vaccines means that there are few BCG-free populations in the world that would be suitable for trials of future antimycobacterial vaccines . These facts have implications with regard to strategies for the development and testing of new vaccines against mycobacterial diseases. Protein Eng, 1989 Mar, 2(6), 473 - 80 Protein secretion controlled by a synthetic gene in Escherichia coli; Blanchin-Roland S et al.; The inability of Escherichia coli to secrete proteins in growth medium is one of the major drawbacks in its use in genetic engineering . A synthetic gene, homologous to the one coding for the kil peptide of pColE1, was made and cloned under the control of the lac promoter, in order to obtain the inducible secretion of homologous or heterologous proteins by E . coli . The efficiency of this synthetic gene to promote secretion was assayed by analysing the production and secretion of two proteins, the R-TEM1 beta-lactamase, and the alpha-amylase from Bacillus licheniformis . This latter protein was expressed in E . coli from its gene either on the same plasmid as the kil gene or on a different plasmid . The primary effect of the induction of the kil gene is the overproduction of the secreted proteins . When expressed at a high level, the kil gene promotes the overproduction of all periplasmic proteins and the total secretion in the culture medium of both the beta-lactamase or the alpha-amylase . This secretion is semi-selective for most periplasmic proteins are not secreted . The kil peptide induces the secretion of homologous or heterologous proteins in two steps, first acting on the cytoplasmic membrane, then permeabilizing the outer membrane . This system, which is now being assayed at the fermentor scale, is the first example of using a synthetic gene to engineer a new property into a bacterial strain. Pediatr Infect Dis J, 1989 Mar, 8(3), 155 - 9 Cerebrospinal fluid interleukin 1-beta and tumor necrosis factor concentrations and outcome from neonatal gram-negative enteric bacillary meningitis; McCracken GH Jr et al.; Concentrations of interleukin 1-beta and tumor necrosis factor in cerebrospinal fluid (CSF) of 42 neonates with Gram-negative enteric bacillary meningitis were determined and correlated with outcome and other CSF indices . At the time of diagnosis interleukin 1-beta was detected in CSF of 40 infants and peak concentrations correlated significantly with outcome from disease . Days of interleukin 1-beta concentrations of greater than 200 pg/ml or greater than or equal to 20 pg/ml were significantly correlated with days that CSF cultures were positive and that K1 antigen and endotoxin were detected in CSF . Tumor necrosis factor activity was detected in CSF of 25 of 27 infants; there was no correlation between outcome and CSF tumor necrosis factor concentrations . The results provide a possible rationale for therapeutic intervention to improve outcome. Med Hypotheses, 1989 Mar, 28(3), 145 - 9 Cat-scratch disease and the role of the domestic cat: vector, reservoir, and victim? Kirkpatrick CE, Glickman LT. An unidentified coccobacillus has been implicated recently as the agent of Cat-Scratch Disease (CSD) in human beings . Although a history of close contact with a domestic cat is frequently elicited from CSD patients, the exact role of this animal in the epidemiology of the disease remains obscure . Current thinking holds that the cat merely serves as an efficient inoculator of a free-living organism . We believe that the cat is not only an important vector of the CSD bacillus, but it may also serve as the principal reservoir, with the organism occasionally present among the oral flora . Under some circumstances (e.g., immunoincompetency) the CSD bacillus may also infect the lymph nodes of cats, resulting in a disease similar to CSD in human beings. Rev Infect Dis, 1989 Mar-Apr, 11(2), 335 - 7 Cure of bacille Calmette-Guérin vaccination abscesses with erythromycin; Murphy PM et al.; Postvaccination subcutaneous abscess due to bacille Calmette-Guerin (BCG) is an uncommon complication and is especially rare in the United States, where the general population is not vaccinated with BCG . This type of abscess is usually chronic, and optimal therapy has not been defined . Two Americans, a husband and wife, underwent primary BCG vaccination abroad and developed chronic subcutaneous abscesses at the primary inoculation site . Four months after vaccination, Mycobacterium bovis strain BCG was cultured from material aspirated from both lesions . Direct susceptibility studies revealed a minimal inhibitory concentration of less than 3.0 micrograms of erythromycin/mL for both isolates . Erythromycin was given orally to the husband and wife for 3 and 4 weeks, respectively, during which time complete healing occurred in both cases. Appl Environ Microbiol, 1989 Mar, 55(3), 568 - 72 Identification of two distinct Bacillus circulans xylanases by molecular cloning of the genes and expression in Escherichia coli; Yang RC et al.; Two genes coding for xylanase synthesis in Bacillus circulans were cloned and expressed in Escherichia coli . After digestion of genomic DNA from Bacillus circulans with EcoRI and PstI, the fragments were ligated into the corresponding sites of pUC19 and transformed into Escherichia coli . Restriction enzyme mapping of the two inserts coding for xylanase activity indicated distinctly different nucleotide sequences . Cross-hybridization assays confirmed the absence of sequence homology between the two genes . In vitro transcription-translation assays indicated that the cloned genes encoded for proteins with molecular weights of 22,000 and 59,000 . The gene products displayed different substrate specificities . The 22,000-dalton enzyme readily hybrolyzed aspeen, larchwood, and oat spelt xylans, whereas the second was unable to extensively depolymerize oat spelt xylan and resulted in very limited reducing sugar release from any of the xylan substrates tested . Both of the xylanases had isoelectric points of approximately 9.0. J Bacteriol, 1989 Mar, 171(3), 1705 - 11 Molecular cloning and expression in Escherichia coli of the Bacillus licheniformis bacitracin synthetase 2 gene; Ishihara H et al.; The structural genes for the entire bacitracin synthetase 2 (component II) and for a part of the putative bacitracin synthetase 3 (component III) from Bacillus licheniformis ATCC 10716 were cloned and expressed in Escherichia coli . A cosmid library of B . licheniformis DNA was constructed . The library was screened for the ability to produce bacitracin synthetase by in situ immunoassay using anti-bacitracin synthetase antiserum . A positive clone designated B-15, which has a recombinant plasmid carrying about a 32-kilobase insert of B . licheniformis DNA, was further characterized . Analysis of crude cell extract from B-15 by polyacrylamide gel electrophoresis and Western blotting (immunoblotting) showed that the extract contains two immunoreactant proteins with high molecular weight . One band with a molecular weight of about 240,000 comigrates with bacitracin synthetase 2; the other band is a protein with a molecular weight of about 300,000 . Partial purification of the gene products encoded by the recombinant plasmid by gel filtration and hydroxyapatite column chromatography revealed that one gene product catalyzes L-lysine- and L-ornithine-dependent ATP-PPi exchange reactions which are characteristic of bacitracin synthetase 2, and the other product catalyzes L-isoleucine-, L-leucine, L-valine-, and L-histidine-dependent ATP-PPi exchange activities, suggesting the activities of a part of bacitracin synthetase 3 . Subcloning experiments indicated that the structural gene for bacitracin synthetase 2 is located near the middle of the insert. J Surg Oncol, 1989 Mar, 40(3), 205 - 13 Adjuvant chemoimmuno- and immunotherapy in Dukes' stage B2 and C colorectal carcinoma: a 7-year follow-up analysis; Abdi EA et al.; A prospectively randomised controlled clinical trial of adjuvant therapy was undertaken, at a single-centre, population-based cancer institute, in patients with Dukes' stages B2 and C colorectal carcinoma after curative surgery . Between 1976 and 1983, 253 patients were randomised to either control (no further therapy after surgery), immunotherapy (oral bacille Calmette-Guerin {BCG} 120 mg once a month) for 5 years or chemoimmunotherapy (oral BCG as above with methyl-cyclohexyl-chloroethyl nitrosourea {meCCNU} 130 mg/m2 on day 1 and 5-fluorouracil {5-FU} 325 mg/m2/day on days 1-5 and 375 mg/m2/day on days 36-40) repeated every 10 weeks for 8 cycles . The median follow-up of patients is now 6.95 years . Of the control, immunotherapy, and chemoimmunotherapy groups 22.35%, 39.28%, and 28.57%, respectively, have relapsed . The log-rank analysis of results shows no disease-free or overall survival advantage for patients receiving adjuvant therapy compared with the control group . Patients receiving adjuvant immunotherapy for stage B2 appear to have a significantly inferior disease-free survival compared with other groups, but their overall survival is similar . There are no significant differences in disease-free or overall survival in the three groups of patients with stage C tumour . Of 82 patients dying, 78.05% died of progressive colorectal carcinoma, 13 patients developed a second malignancy; the remainder died of seemingly unrelated causes. J Invertebr Pathol, 1989 Mar, 53(2), 251 - 9 Variation in binding of Bacillus sphaericus toxin and wheat germ agglutinin to larval midgut cells of six species of mosquitoes; Davidson EW; Bacillus sphaericus toxin labeled with fluorescein isothiocyanate was readily ingested by Culex pipiens, Aedes aegypti, Anopheles stephensi, Anopheles gambiae, Anopheles quadrimaculatus, and Anopheles albimanus larvae . Fluorescent toxin bound to the luminal cell surface in discrete regions of the posterior midgut and gastric caecum in C . pipiens . In Anopheles spp., toxin bound in a variable pattern to these structures and central and anterior midgut as well . The toxin did not bind to midgut cells of A . aegypti . The toxin was internalized in bright fluorescent vesicles in C . pipiens, but was not internalized in Anopheles spp . and appeared to be weakly bound in these larvae, leaking rapidly from the gut surface . The lectin, wheat germ agglutinin, which interferes with binding of the B . sphaericus toxin, bound to the posterior midgut and gastric caecum of all species, but was not internalized . These results suggest that the sugar moiety of the receptor is not solely responsible for specificity of this toxin, and that binding to Culex spp . midgut cells may be highly specific and of high affinity, whereas binding to Anopheles spp . cells may be nonspecific and/or of low affinity. Mol Biol (Mosk), 1989 Mar-Apr, 23(2), 463 - 72 {Cloning and comparative characteristics of genes coding two structurally distant delta-endotoxins of Bacillus thuringiensis var . galleriae and kurstaki}; Osterman AL et al.; Representative total DNA libraries of Bac . thuringiensis var . kurstaki (strain Dipel) and galleriae (strain 11-67) have been constructed on the basis of phasmid vector lambda pSL5 . Recombinant phasmid clones, carrying delta-endotoxin-coding genes of both subspecies have been isolated by means of immunoenzyme screening . Restriction mapping and partial nucleotide sequence determination have demonstrated that phasmid lambda pOC2, isolated from var . kurstaki DNA library, contains the complete delta-endotoxin-coding gene, identical to the one, described by Schnepf H.E . et al . J . Biol . Chem . 1985 . V . 260 . P . 6264 . Recombinant phasmids lambda pOC10 and 11 have been shown to contain the full-sized gene, coding delta-endotoxin of Bac . thuringiensis var . galleriae . The protein products of the cloned genes have been characterized by Western-blot analysis and bioassays . The absence of substantial homology of two genes, evidenced by Southern-blot hybridisation, correlates with sufficiently big differences in biological specificity of the corresponding proteins . This is in accordance with our previous data on N-terminal amino acid sequence determination of delta-endotoxins of those subspecies of Bac . thuringiensis. Anticancer Res, 1989 Mar-Apr, 9(2), 391 - 3 Intrapleural BCG in postsurgical stage I non-small cell lung cancer; Macchiarini P et al.; In a randomized trial comparing intrapleural bacillus Calmette-Guerin (BCG) with placebo in postsurgical stage I non-small cell lung cancer, BCG-treated patients (n = 17) showed a non-significant overall improvement of 5-year and median survival and disease-free interval, but an increased rate of recurrence of cancer compared with control patients (n = 17). J Immunol, 1989 Mar 1, 142(5), 1696 - 701 Suppression of monocyte oxidative response by phenolic glycolipid I of Mycobacterium leprae; Vachula M et al.; Mycobacterium leprae synthesizes a unique phenolic glycolipid (PGL-I) in abundant quantities . We studied the effect of PGL-I on the generation of superoxide anion (O2-) by stimulated human monocytes . Peripheral blood monocytes pretreated with PGL-I released less O2- when stimulated with M . leprae than did control monocytes . Monocytes pretreated with dimycocerosyl phthiocerol, mycoside A of Mycobacterium kansasii, or mycoside B of Mycobacterium microti, on the other hand, released O2- in quantities comparable to control monocytes in response to M . leprae stimulation . Monocyte O2- release in response to other stimuli of the oxidative metabolic burst, such as PMA, zymosan, Mycobacterium bovis Bacille Calmette-Guerin, or M . kansasii, was unaffected by lipid pretreatment . These findings demonstrate that PGL-I has a direct effect on monocyte O2- generation in response to M . leprae and suggest that PGL-I is a modulator of phagocytic cell function. J Cell Biochem, 1989 Mar, 39(3), 315 - 25 Phosphatidylinositol-specific phospholipase C from Bacillus cereus: improved purification, amino acid composition, and amino-terminal sequence; Volwerk JJ et al.; Phosphatidylinositol-specific phospholipase C was purified in a 27% yield from the culture medium of Bacillus cereus by a combination of ammonium sulfate precipitation and ion-exchange and hydrophobic interaction chromatography . The purified enzyme was free of other phospholipase C-type activities and exhibited a high specific activity of approximately 1,300 units/mg . Amino acid composition analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated a molecular weight of about 35 kDa . The sequence of the first 29 N-terminal amino acids was also determined. FEBS Lett, 1989 Feb 27, 244(2), 259 - 62 A cytolytic delta-endotoxin from Bacillus thuringiensis var . israelensis forms cation-selective channels in planar lipid bilayers; Knowles BH et al.; In order to determine the mechanism of action of the 27 kDa mosquitocidal delta-endotoxin of Bacillus thuringiensis var . israelensis we have studied its effects on the conductance of planar lipid bilayers . The toxin formed cation-selective channels in the bilayers, permeable to K+ and Na+ but not to N-methylglucamine or Cl-, showing very fast, cooperative opening and closing . Channel opening was greatly reduced in the presence of divalent cations (Ca2+, Mg2+) and the effect was reversed when these ions were removed . These results are consistent with our proposal that B . thuringiensis toxins act by a mechanism of colloid-osmotic lysis. J Biol Chem, 1989 Feb 25, 264(6), 3200 - 5 Structure and properties of malic enzyme from Bacillus stearothermophilus; Kobayashi K et al.; The malic enzyme (EC 1.1.1.38) gene of Bacillus stearothermophilus was cloned in Escherichia coli, and the enzyme was purified to homogeneity from the E . coli clone . In addition to the NAD(P)-dependent oxidative decarboxylation of L-malate, the enzyme catalyzes the decarboxylation of oxalacetate . The enzyme is a tetramer of Mr 200,000 consisting of four identical subunits of Mr 50,000 . The pH optima for malate oxidation and pyruvate reduction are 8.0 and 6.0, respectively; and the optimum temperature is 55 degrees C . The enzyme strictly requires divalent metal cations for its activity, and the activity is enhanced 5-7 times by NH4+ and K+ . Kinetic study shows that the values of the dissociation constant of the enzyme-coenzyme complex are 77 microM for NAD and 1.0 mM for NADP, indicating that the enzyme has a higher affinity for NAD than for NADP . The nucleotide sequence of the gene and its flanking regions was also found . A single open reading frame of 1434 base pairs encoding 478 amino acids was concluded to be that for the malic enzyme gene because the amino acid composition of the enzyme and the sequence of 16 amino acids from the amino terminus of the enzyme agreed well with those deduced from this open reading frame. J Mol Biol, 1989 Feb 20, 205(4), 729 - 35 Structural and kinetic bases for the recognition of tRNATyr by tyrosyl-tRNA synthetase; Labouze E et al.; The aminoacylation of transfer RNA is a key step of translation since it relates amino acids to anticodons . To understand how the tyrosyl-tRNA synthetase (TyrTS) from Bacillus stearothermophilus recognizes tRNA(Tyr), we constructed 14 new mutant TyrTS by site-directed mutagenesis, determined their kinetic properties and used these and previous data to construct a detailed structural model of the complex between TyrTS and the acceptor arm of tRNA(Tyr) . In the model Arg207, Lys208, Asn 146 and Glu 152 interact with phosphate groups . A contact between guanine 1 and Trp 196 is unspecific . Adenine 73, the fourth base from the 3' end, is specifically recognized through Trp 196 and the main-chain carbonyl of Ala150 . At the active site, adenine 76 might interact with Lys82 and Arg86 . There is a tight complementarity in shape between the tRNA and the synthetase . TyrTS and tRNA(Tyr) form an additional contact, in the vicinity of adenine 73, when their complex goes from the initial state to the transition state . The rate of aminoacylation, through the precise recognition of adenine 73, could thus be an important factor of discrimination by TyrTS among tRNAs. Cancer, 1989 Feb 15, 63(4), 667 - 70 Ethylchlorformate polymerized tumor protein immunotherapy of the murine bladder tumor; Mori K et al.; Using murine transplantable transitional cell carcinoma (MBT2), the effect of ethylchlorformate (ECF) polymerized tumor protein was compared with that of bacillus Calmette-Guerin (BCG) . Seventy-five C3H/He mice were challenged with an intradermal inoculation of 5 X 10(5) viable MBT2 tumor cells and divided into five groups . Each group was intradermally administered with 0.01 mg of ECF (low ECF), 0.25 mg of ECF (high ECF), 0.1 mg of ECF and 10(6) CFU BCG (ECF/BCG), 10(6) CFU of BCG alone or normal saline (control) weekly for 10 weeks . The mean survival rate for the treatment groups was 64 to 73 days and significantly longer than that for the control group (P less than 0.001, Savage) . The incidence of biologically active tumor progression was significantly less for the treatment groups (low ECF, 53%; high ECF, 33%; ECF/BCG, 7%; BCG, 27%) compared with the control group (87%; P less than 0.5, chi-square . The mean rate of tumor growth was significantly lower for all treatment groups than for the control group (P less than 0.001, ANOVA and SNK), and the ECF/BCG group had the lowest growth rate despite a higher incidence of local granulomatous reaction . In this study, immunotherapy significantly prolonged the survival rate, decreased the incidence of biologically active tumor progression, and slowed the rate of tumor growth . The combination of ECF polymerized tumor protein and BCG had the greatest effect, suggesting that the effect of the vaccine was increased with BCG. FEBS Lett, 1989 Feb 13, 244(1), 31 - 3 Location of enzymatic and DNA-binding domains on E . coli protease La; Baker ME; Escherichia coli protease La is an ATP-dependent enzyme that has a DNA-binding site . The locations of the enzymatic and DNA-binding sites are not known . We report that a 75-residue segment at the carboxy-terminus of the protease La is similar to part of Bacillus licheniformis beta-lactamase, a serine enzyme . The comparison score is 8.2 standard deviations higher than that obtained with 10,000 comparisons of randomized sequences of these segments . The probability of obtaining such a score by chance is 1.2 x 10(-16) . We also find that a 107-residue segment in the amino-terminus half of protease La is similar to part of the sopB protein, a DNA-binding protein of the plasmid F of E . coli . The comparison score for these segments is 8 standard deviations (P = 6 x 10(-16)) . These strong amino acid sequence similarities suggest the locations of the catalytic serine and the DNA-binding domains of protease La. Nucleic Acids Res, 1989 Feb 11, 17(3), 979 - 97 Cloning the BamHI restriction modification system; Brooks JE et al.; BamHI, a Type II restriction modification system from Bacillus amyloliquefaciensH recognizes the sequence GGATCC . The methylase and endonuclease genes have been cloned into E . coli in separate steps; the clone is able to restrict unmodified phage . Although within the clone the methylase and endonuclease genes are present on the same pACYC184 vector, the system can be maintained in E . coli only with an additional copy of the methylase gene present on a separate vector . The initial selection for BamHI methylase activity also yielded a second BamHI methylase gene which is not homologous in DNA sequence and hybridizes to different genomic restriction fragments than does the endonuclease-linked methylase gene . Finally, the interaction of the BamHI system with the E . coli Dam and the Mcr A and B functions, have been studied and are reported here. Biochemistry, 1989 Feb 7, 28(3), 945 - 52 Conformational states of beta-lactamase: molten-globule states at acidic and alkaline pH with high salt; Goto Y et al.; We present evidence that beta-lactamase is close to fully unfolded (i.e., random coil conformation) at low ionic strength at the extremes of pH and that the presence of salt causes a cooperative transition to a conformation with the properties of a molten globule, namely, a compact state with native-like secondary structure but disordered side chains (tertiary structure) . The conformation of beta-lactamase I from Bacillus cereus was examined over the pH 1.5-12.5 region by circular dichroism (CD), tryptophan fluorescence, dynamic light scattering, and 1-anilino-8-naphthalenesulfonate (ANS) binding . Under conditions of low ionic strength (I = 0.05) beta-lactamase was unfolded below pH 2.5 and above pH 11.5, on the basis of the far-UV and near-UV CD and tryptophan fluorescence . However, at high ionic strength and low pH an intermediate conformation (state A) was observed, with a secondary structure content similar to that of the native protein but a largely disordered tertiary structure . The transition from the unfolded state (U) to state A induced by KCl was cooperative and had a midpoint at 0.12 M KCl (I = 0.17 M) at pH 1.6 . A similar conformation (state B) was observed at high pH and high ionic strength . The transition from the alkaline U state to state B induced by KCl at pH 12.2 was cooperative and had a midpoint at 0.6 M KCl (I = 0.65 M) . Light scattering measurements showed that state B was compact although somewhat expanded compared to the N state . The compactness of state A could not be determined due to its strong propensity to aggregate.(ABSTRACT TRUNCATED AT 250 WORDS) J Mol Biol, 1989 Feb 5, 205(3), 619 - 21 Crystals of wild-type, mutated, derivatized and complexed 50 S ribosomal subunits from Bacillus stearothermophilus suitable for X-ray analysis; Mussig J et al.; Three-dimensional single crystals of wild-type and mutated 50 S ribosomal subunits from Bacillus stearothermophilus, as well as crystals of reconstituted subunits containing heavy-atom clusters and complexes of these subunits with tRNA and a short nascent polypeptide chain, were grown from polyethylene glycol in the presence of salts at low concentrations . Within experimental error, all these crystals are isomorphous, packed with monoclinic symmetry (C2) in unit cells of a = 300 A, b = 546 A, c = 377 (+/- 1%) A and beta = 112 degrees . Using synchrotron radiation at 85 to 100 K they diffract to 11 A resolution and can be irradiated for hours without disintegrating, so that a complete data set could be collected from a single crystal. J Biol Response Mod, 1989 Feb, 8(1), 37 - 52 Melanoma patient antibody responses to melanoma tumor-associated antigens defined by murine monoclonal antibodies; Seigler HF et al.; Sera from melanoma patients undergoing immunization with three distinct immunogens were evaluated for development of IgG antibody specific for the melanoma tumor-associated antigens (TAAs) GD3 and 250-kd glycoprotein (high molecular weight; HMW) . Of 99 patients receiving either irradiated allogeneic melanoma cells admixed with bacillus Calmette-Guerin, vaccinia viral oncolysate melanoma cell membranes, or cholesterol hemisuccinate-modified melanoma cells, 12 were determined to have responded against GD3 and 17 against the HMW TAA . This reactivity was measured using both direct binding to purified TAA and specific inhibition of binding of murine monoclonal antibodies R24 and Me-1-14 for GD3 and HMW TAA, respectively . Preimmune sera from these patients did not react with these TAAs, nor did preimmune sera or follow-up sera from melanoma patients electing not to receive immunotherapy . These results suggest that melanoma patients can be immunized against these TAAs as presented on the melanoma cell membrane in an immunotherapy setting and that immunization using purified TAAs might likewise be feasible. Int J Radiat Oncol Biol Phys, 1989 Feb, 16(2), 459 - 63 Long-term survival and patterns of failure after postoperative radiation therapy for subtotally resected rectal adenocarcinoma; Schild SE et al.; Between 1977 and 1984, 17 patients received external beam irradiation after subtotal resection of rectal carcinoma . Ten patients had microscopic residual disease and 7 had gross residual disease . In the group with microscopic residual disease, 4 had tumor cut through with pathologically involved margins, 5 had adjacent unresected structures that were biopsy positive, and 1 had tumor spillage into the pelvis . The patients with gross residual disease were noted by the surgeon to have visible tumor after maximal debulking . Nine of 17 cases had involved pelvic lymph nodes . Radiation was administered to the pelvis with 4, 6, or 10 MV photons . Doses ranged from 40 to 60 Gy, with a median dose of 50 Gy given at 1.8 to 2.0 Gy per fraction, 5 days per week . Three patients received bacillus Calmette-Guerin (BCG), 2 received 5-fluorouracil (5-FU), and 1 received hycanthone . Thirteen of the 17 patients (76%) experienced local failure and, of these, 10 also developed distant disease . No patients developed distant metastasis in the absence of local failure . Local control was achieved in 3 of 10 patients (30%) with microscopic residual and 1 of 7 (14%) with gross residual . Four of the 17 patients (24%) have remained free of disease for greater than 5 years . External beam irradiation is capable of producing long-term survival and local control in a minority of patients with rectal cancer after subtotal resection . Investigation of more aggressive forms of therapy such as the addition of intraoperative irradiation, brachytherapy, radiation dose modifiers, and chemotherapy is warranted. J Bacteriol, 1989 Feb, 171(2), 965 - 74 Two highly related insecticidal crystal proteins of Bacillus thuringiensis subsp . kurstaki possess different host range specificities; Widner WR et al.; Two genes encoding insecticidal crystal proteins from Bacillus thuringiensis subsp . kurstaki HD-1 were cloned and sequenced . Both genes, designated cryB1 and cryB2, encode polypeptides of 633 amino acids having a molecular mass of ca . 71 kilodaltons (kDa) . Despite the fact that these two proteins display 87% identity in amino acid sequence, they exhibit different toxin specificities . The cryB1 gene product is toxic to both dipteran (Aedes aegypti) and lepidopteran (Manduca sexta) larvae, whereas the cryB2 gene product is toxic only to the latter . DNA sequence analysis indicates that cryB1 is the distal gene of an operon which is comprised of three open reading frames (designated orf1, orf2, and cryB1) . The proteins encoded by cryB1 and orf2 are components of small cuboidal crystals found in several subspecies and strains of B . thuringiensis; it is not known whether the orf1 or cryB2 gene products are present in cuboidal crystals . The protein encoded by orf2 has an electrophoretic mobility corresponding to a molecular mass of ca . 50 kDa, although the gene has a coding capacity for a polypeptide of ca . 29 kDa . Examination of the deduced amino acid sequence for this protein reveals an unusual structure which may account for its aberrant electrophoretic mobility: it contains a 15-amino-acid motif repeated 11 times in tandem . Escherichia coli extracts prepared from cells expressing only orf1 and orf2 are not toxic to either test insect. Antibiot Khimioter, 1989 Feb, 34(2), 129 - 33 {Influence of Bacillus intermedius RNAase on thymus cell composition and T-lymphocyte differentiation in an experiment}; Kurinenko BM et al.; It was shown that exposure of mice AKR and (CBA X C57B1/6)F1 to Bacillus intermedius RNAase and its derivative selectively inactivated by the active centre histidine stimulated T-lymphocyte maturation . With using bone marrow and spleen lymphocytes of congenitally thymus-free mice it was revealed that this effect of RNAase was defined by induction of Thy-1 antigen of the lymphocyte plasmic membrane and not associated with homology of the enzyme primary structure and the alpha 1-thymosin structure. Nippon Hinyokika Gakkai Zasshi, 1989 Feb, 80(2), 167 - 74 {Analyses of the effects of intravesical Bacillus Calmette-Guerin (Tokyo 172 strain) therapy of superficial bladder cancer}; Akaza H et al.; In this paper, we summarize the results of a multicentric controlled study of induction treatment with intravesical BCG in patients with superficial bladder cancer (Ta, T1) or CIS . The BCG Tokyo 172 strain (Japan BCG K.K.) was given for 8 weeks, as a rule, in a dose of either 80 mg or 120 mg in 40 ml of saline instilled into the bladder . Sixty-six (90.5%) of the 73 patients registered in the study were evaluable for response . High response rates were obtained in the patients with Ta or T1 tumors, irrespective of the treatment dose; 65.4% CR, 27.3% PR and 7.3% NC for all . The number of CIS patients is too small to draw any conclusions, but 54.5% CR, 36.4% PR and 9.1% NC were obtained . Several factors were evaluated in relation to the tumor ablative effects . The CR rate was significantly higher for tumors less than 1 cm in size compared with tumors 1-3 or larger in size, although the response rates, including CR and PR, were very similar irrespective of the size . Other factors did not contribute to provoke a significant difference . Japanese elder people show a high positive rate of PPD reaction . Thus, in our study, 78% of the patients showed positive PPD reaction . In this situation, no significant relation between PPD reaction and BCG efficacy was obtained . In addition, OKT8, OKT7, OKT4, OKT4/OKT8 ratio, Leu7, ADCC activity and NK activity did not show any correlation with tumor ablative effect of BCG.(ABSTRACT TRUNCATED AT 250 WORDS) Jpn J Exp Med, 1989 Feb, 59(1), 9 - 15 Production of Tyzzer's disease in rats by ingestion of bacterial spores; Itoh T et al.; Tyzzer's disease was produced in rats by peroral inoculation with spores of Bacillus piliformis (Tyzzer's organism) of rat origin . After ingestion of 10(6) spores necrotized lesions with intracellular bacterial propagation were seen in the intestines, liver and heart on days 2 to 14 postinoculation (p.i.) . A number of B . piliformis were present within enterocytes of the cecum and colon . Infected cells were also seen in the liver, myocardium and intestinal muscle layers on days 3 to 7 p.i . Infective spores were found to be shed in feces during 3 to 10 days p.i. Appl Environ Microbiol, 1989 Feb, 55(2), 500 - 2 Isolation of phenol-degrading Bacillus stearothermophilus and partial characterization of the phenol hydroxylase; Gurujeyalakshmi G et al.; Bacillus stearothermophilus BR219, isolated from river sediment, degraded phenol at levels to 15 mM at a rate of 0.85 mumol/h (4 x 10(6) cells) . The solubilized phenol hydroxylase was NADH dependent, exhibited a 55 degrees C temperature optimum for activity, and was not inhibited by 0.5 mM phenol. Res Microbiol, 1989 Feb, 140(2), 143 - 50 Antagonism between dapsone and rifampicin in experimental Mycobacterium leprae infections in mice; Millan JP et al.; In experimental infections of normal mice with Mycobacterium leprae, the bactericidal activity of four consecutive weekly doses of rifampicin (RMP) was suppressed when this treatment was preceded, for one month, by daily administration of dapsone (DDS), and then the latter . Up until now, it has been impossible to detect this antagonism between the action of RMP and DDS, since all studies involved the simultaneous administration of these two drugs, and such a phenomenon would therefore have been masked by the rapid and potent action of RMP . Previous clinical observations suggest that such a delayed antagonistic effect may also occur in humans . The demonstration of this antagonism between RMP and DDS raises the problem of the long-term efficacy of therapeutic regimens currently used in leprosy and that of the role of DDS in induction of bacillary persistence . It is suggested that this particular methodology, the delayed combination of RMP with a less active drug, should be applied to the study of other drug combinations recommended in the treatment of leprosy. Indian J Biochem Biophys, 1989 Feb, 26(1), 48 - 51 Modification of glutamine synthetase in Bacillus brevis AG 4; Tiwari RP et al.; The various forms of glutamine synthetase obtained from Bacillus brevis have been found to be antigenically identical . Alkaline phosphatase treatment of the fast moving form (GS4) reduced the electrophoretic mobility of the enzyme . Radiolabelling and autoradiographic studies have also indicated that 32P-incorporation is high in the form depicting high Rm value . Thus, it appears that these forms arise due to covalent modification of the enzyme involving a phosphate group. Indian J Biochem Biophys, 1989 Feb, 26(1), 43 - 7 Multiple forms of glutamine synthetase in Bacillus brevis AG 4; Tiwari RP et al.; Glutamine synthetase in Bacillus brevis AG 4, a Gram-positive spore forming bacteria, has been found to exist in multiple molecular forms . It was purified to electrophoretic homogeneity by single-step Blue Sepharose affinity chromatography . The native enzyme has a molecular weight of 600,000 with subunits of 50,000 . The enzyme samples purified from different stages of growth differed in Mg2+ sensitivity and other kinetic properties . Four different enzyme samples selected on the basis of Mg2+ sensitivity showed distinct mobilities at pH 6.3 on PAGE using discontinuous buffer system . A correlation amongst Mg2+ sensitivity, electrophoretic mobility, and kinetic properties was highly suggestive of multiple forms of glutamine synthetase in Bacillus brevis arising due to modification. Appl Environ Microbiol, 1989 Feb, 55(2), 294 - 7 Carbohydrate metabolism in the mosquito pathogen Bacillus sphaericus 2362; Russell BL et al.; Bacillus sphaericus 2362 is pathogenic for mosquito larvae and is being considered for large-scale production as a larvicide . The inability of the bacteria to metabolize carbohydrates requires that they be grown on proteinaceous media . This bacterium was found to be unable to transport glucose or sucrose into the cell, and it lacked glucokinase and hexokinase activity . In addition, it lacked phosphoglucose isomerase, phosphofructokinase, and glucose 6-phosphate dehydrogenase, which are early enzymes of the Embden-Myerhof-Parnas and hexose monophosphate pathways . The presence of other enzymes in these pathways was indicated by assay, by the metabolism of glycerol to acetate, and by growth on acetate and gluconate as sole carbon sources . Critical enzymes of the Entner-Doudoroff pathway were also shown to be absent. J Bacteriol, 1989 Feb, 171(2), 1118 - 25 Mechanism of L-glutamate transport in membrane vesicles from Bacillus stearothermophilus; de Vrij W et al.; In the presence of electrochemical energy, several branched-chain neutral and acidic amino acids were found to accumulate in membrane vesicles of Bacillus stearothermophilus . The membrane vesicles contained a stereo-specific transport system for the acidic amino acids L-glutamate and L-aspartate, which could not translocate their respective amines, L-glutamine and L-asparagine . The transport system was thermostable (Ti = 70 degrees C) and showed highest activities at elevated temperatures (60 to 65 degrees C) . The membrane potential or pH gradient could act as the driving force for L-glutamate uptake, which indicated that the transport process of L-glutamate is electrogenic and that protons are involved in the translocation process . The electrogenic character implies that the anionic L-glutamate is cotransported with at least two monovalent cations . To determine the mechanistic stoichiometry of L-glutamate transport and the nature of the cotranslocated cations, the relationship between the components of the proton motive force and the chemical gradient of L-glutamate was investigated at different external pH values in the absence and presence of ionophores . In the presence of either a membrane potential or a pH gradient, the chemical gradient of L-glutamate was equivalent to that specific gradient at different pH values . These results cannot be explained by cotransport of L-glutamate with two protons, assuming thermodynamic equilibrium between the driving force for uptake and the chemical gradient of the substrate . To determine the character of the cotranslocated cations, L-glutamate uptake was monitored with artificial gradients . It was established that either the membrane potential, pH gradient, or chemical gradient of sodium ions could act as the driving force for L-glutamate uptake, which indicated that L-glutamate most likely is cotranslocated in symport with one proton and on sodium ion. Mol Microbiol, 1989 Feb, 3(2), 229 - 38 Nucleotide sequence and analysis of the N-terminal coding region of the Spodoptera-active delta-endotoxin gene of Bacillus thuringiensis aizawai 7.29; Sanchis V et al.; The nucleotide sequence of a 2711bp DNA segment which contains the N-terminal coding sequence and the 5' flanking region of a crystal protein gene (bta) from Bacillus thuringiensis subsp . aizawai 7.29 has been determined . The coding region encodes an 824 amino-acid polypeptide corresponding to a carboxy-terminally truncated delta-endotoxin specifically active against the cotton leaf worm Spodoptera littoralis . Comparison of the deduced amino acid sequence of the bta gene with that of the 4.5, 5.3 and 6.6 kb classes of lepidopteran-active delta-endotoxins revealed that the Bta sequence contains a very high level of amino acid substitutions in the N-terminal part of the protoxin molecule . The substitutions are grouped in several highly variable segments separated by highly conserved regions . These conserved domains are also present in the dipteran- and coleopteran-active delta-endotoxins . The control region of the bta gene shows considerable DNA identity with the control regions of the other lepidopteran-active genes . Deletions of the 3' region of the gene were carried out and the toxic fraction of the bta delta-endotoxin was identified with the N-terminal half of the molecule. Am J Trop Med Hyg, 1989 Feb, 40(2), 186 - 94 Depression of hydrogen peroxide dependent killing of schistosomula in vitro by peritoneal exudate cells from Schistosoma mansoni infected mice; Smith JM et al.; Peritoneal exudate cells from mice infected with Schistosoma mansoni (S-PEC) can kill a small proportion of schistosomula in vitro in the presence of immune serum . S-PEC produce a low level of respiratory burst . However, schistosomula mortality in their presence is not reduced when exogenous antioxidants are added, suggesting that with S-PEC, oxidative killing may not be important . Hydrogen peroxide (H2O2) and superoxide production by S-PEC, and cells from Bacillus Calmette-Guerin (BCG) and thioglycollate (THGL) injected mice, nonspecifically stimulated with opsonized zymosan, were measured . Levels of H2O2 produced by S-PEC were significantly lower than BCG or THGL PEC, and were below the threshold for schistosomula killing . This correlated with lower levels of cell-mediated killing of schistosomula in vitro by S-PEC than by BCG-PEC . Superoxide levels, however, were similar between the 3 cell populations . It therefore appears that the efficiency of PEC to kill schistosomules in vitro correlates with H2O2 rather than superoxide levels . It was found that there was a sharp concentration threshold in H2O2 mediated killing of schistosomula . A depression in the levels of H2O2 produced may be a mechanism by which the parasite can partially evade the host immune system. Zh Mikrobiol Epidemiol Immunobiol, 1989 Feb, (2), 8 - 11 {Change in the ultrastructure of the cell wall of Bacillus cereus, determined by RP4 MUCTS62 plasmid}; Goderdzishvili MG et al.; The influence of plasmid RP4 Mucts62, heterologous for B . cereus, on the growth rate of B . cereus strains GA 682 and 319 obtained in our earlier experiments and on changes in the ultrastructure of their cell walls in comparison with B . cereus initial strains GP 7 and DSM 318 has been studied . Plasmid RP4 Mucts62 with a wide spectrum of action has been found not only to determine the functional signs of resistance to antibiotics and thermal sensitivity in the heterologous host, but also to take part in the morphological organization of the cell surface structure, and in particular in the structure of the S-layer . B . cereus strains containing plasmid RP4 Mucts62 are characterized by slower growth rate and cell fragility. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1989 Feb, 187(3), 244 - 53 Microbiological profile of selected samples of "Al-Kohl" eye cosmetics in northern Jordanian provinces before and after use; Abdelaziz AA et al.; Fifty items of Al-Kohl collected from northern Jordanian provinces representing: 20 unopened ready to use samples purchased from retail outlets, 20 in-use samples obtained from ladies of different socioeconomic standards and 10 samples of the original stones (used for Al-Kohl preparation) were examined for their microbial contents . Ready to use and in-use samples were much more contaminated than the original stones . On sterility testing, more than 85% of the unused and in-use samples were contaminated with bacteria and or fungi comparing to 50% of the original stone samples . Quantitatively, 90% of the original stones contained less than 100 bacterial or fungal cells/g and the other remaining 10% were in the range of 10(2)-10(3) cfu/g either for bacterial or fungal counts . The level and distribution of the viable microbial counts in unused and in-use samples were comparable and much higher than original stones . More than 70% and 20% of those items contained more than 100 cfu/g of bacteria and fungi respectively . Among those samples 20% and 5% were heavily contaminated (contain more than 10(4) cfu/g) with bacteria and fungi respectively . Coliform bacteria in a number of 100 cfu/g or more were recovered from 10% of the unused and 20% of the in-use samples, none were recovered from original stones . The results of qualitative tests for identification of isolated microorganisms showed that 7 different species of Bacillus were found in the 50 examined samples . Approximately 50% of the examined samples contained Bacillus spp., Staphylococcus spp., Pseudomonas spp., P . vulgaris, S . marcescens were recovered from unused and in-use samples in different percentages, none from original stones, some of the detected Staphylococcus were aureus type and also one isolate of P . aeruginosa was detected in one of the in-use samples . The relationship between the detected level of microbial contamination in the tested samples with the proposed allowable international limits of contamination as well as the possible sources of contamination and the hygienic implications of using such products by the public were discussed. J Bacteriol, 1989 Feb, 171(2), 1166 - 72 Replication and segregational stability of Bacillus plasmid pBAA1; Devine KM et al.; A cryptic plasmid, pBAA1, was identified in an industrial Bacillus strain . The plasmid is 6.8 kilobases in size and is present in cells at a copy number of approximately 5 per chromosome equivalent . The plasmid has been maintained under industrial fermentation conditions without apparent selective pressure and so is assumed to be partition proficient . The minimal replicon was localized to a 1.4-kilobase fragment which also contains the functions required for copy number control . The very low level of segregational instability of the minimal replicon suggests that it also contains functions involved in plasmid maintenance . Comparison with other plasmids indicates that pBAA1 belongs to the group of small gram-positive plasmids which replicate by a rolling cycle-type mechanism . A sequence was identified which is required for the efficient conversion of the single plus strand to the double-stranded form during plasmid replication . Deletion of this sequence resulted in a low level of segregational plasmid instability. Gene, 1989 Jan 30, 75(1), 145 - 55 Nucleotide sequence determination of the DNA region coding for Bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase and of the flanking DNA regions required for its expression in Escherichia coli; Branlant C et al.; The complete nucleotide sequence of a 3541-base pairs (bp) DNA fragment from Bacillus stearothermophilus able to complement an Escherichia coli glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) mutant (gapD-) has been determined . The B . stearothermophilus gap gene consists of a 1005-bp open reading frame commencing with an ATG start codon and ending with a TAA stop codon . Upstream from the start codon is a strong Shine-Dalgarno sequence typical of Gram-positive bacteria . Only one putative RNA polymerase recognition signal (-35 and -10 regions) is found 1153 bp upstream from the ATG start codon . In vivo utilization of this signal is in agreement with the study of gene expression from different subclones of the original fragment . 57 bp downstream from the TAA stop codon is a structure resembling Rho-independent transcription termination signals . Although B . stearothermophilus GAPDH-coding gene is highly expressed in E . coli, it contains several rare codons for E . coli . The predicted amino acid sequence of the GAPDH enzyme presents several differences with the amino acid sequence previously determined from the protein and is in better agreement with published crystallographic data. Biochim Biophys Acta, 1989 Jan 30, 978(2), 216 - 22 Mechanism of action of Bacillus thuringiensis insecticidal delta-endotoxin: interaction with phospholipid vesicles; Haider MZ et al.; Bacillus thuringiensis (Bt) crystal delta-endotoxin from three subspecies and the product of a cloned crystal protein gene were activated in vitro and their interaction with phospholipid liposomes studied . Despite their diverse spectrum of activity, all these toxins were found to cause a rapid increase in the light scattering of liposome suspensions, which reflects a morphological change in the lipid bilayer . When liposomes loaded with radioactive markers were incubated with B . thuringiensis aizawai IC1 toxin, a relatively rapid release of more than 70% of the trapped markers occurred after an initial lag . Activated Bta IC1 and B . thuringiensis israelensis toxins were shown to bind to phospholipid vesicles . Two of the five conserved domains (D1-D5) detectable in the sequence of a range of Bt toxins are predicted to be highly hydrophobic . It is suggested that these, together with an additional conserved hydrophobic region showing structural homology and two predicted amphiphilic helices, play a major part in the interaction of these toxins with target membranes. J Biol Chem, 1989 Jan 25, 264(3), 1528 - 33 Organization and sequence of the genes coding for the proton-translocating ATPase of Bacillus megaterium; Brusilow WS et al.; We have cloned and sequenced the genes for the subunits of the proton-translocating ATP synthase of Bacillus megaterium QM B1551 . The arrangement of the genes is identical to the arrangement of the same genes (the unc operon) in Escherichia coli . The genes for the Fo subunits immediately precede the genes for the F1 subunits and are themselves preceded by an open reading frame which codes for a protein similar to the E . coli i protein . In contrast to the E . coli ATPase genes, the transcript for these ATPase genes does not include this open reading frame. J Biol Chem, 1989 Jan 5, 264(1), 131 - 5 Site-directed mutagenesis in Bacillus stearothermophilus fructose-6-phosphate 1-kinase . Mutation at the substrate-binding site affects allosteric behavior; Valdez BC et al.; Arg252 of fructose-6-phosphate 1-kinase (PFK) from Bacillus stearothermophilus has been proposed to be involved in the binding of the substrate Fru-6-P . We demonstrate here that mutation of this residue to alanine converts the enzyme to a form with characteristics similar to those of its allosterically tight form . The mutant enzyme exhibits a high affinity for its inhibitor phosphoenolpyruvate (a 68-fold difference compared to wild type) and a dramatically decreased Fru-6-P affinity (1500-fold increase in Km) . It is more sensitive to inhibition by high ATP concentrations than the wild type, and this inhibition is relieved by ADP, GDP, or higher Fru-6-P concentrations . In contrast, mutation of Arg252 to lysine increases the affinity of the enzyme for P-enolpyruvate by only 2-fold and increases its Km for Fru-6-P by only 50-fold . Sigmoidal kinetics with respect to Fru-6-P in the presence of P-enolpyruvate were observed with Hill numbers of 2.2, 2.4, and 1.7 for wild-type B . stearothermophilus PFK and the Arg252 to lysine and to alanine mutations, respectively . Unlike fructose-6-phosphate 1-kinase from Escherichia coli, in the absence of P-enolpyruvate, B . stearothermophilus PFK exhibits a hyperbolic profile with respect to Fru-6-P concentration . B . stearothermophilus PFK is sensitive to inhibition by high ATP concentrations and competitively inhibited by GDP or ADP . Our data indicate that Arg252 of B . stearothermophilus PFK plays a major role in both Fru-6-P binding and allosteric interaction between the subunits . However, this residue does not seem to participate directly in the catalytic process. Urologe A, 1989 Jan, 28(1), 36 - 9 {BCG in reflux}; Bohle A et al.; Nine patients with radiologically proven vesicorenal reflux (3 bilateral, 4 unilateral, 2 with unilateral double-J stents) received intravesical bacillus Calmette-Guerin (BCG) as prophylactic treatment for recurrent superficial bladder carcinoma . One patient with a double-J stent suffered an attack of high fever, probably following obstruction of the stent . All other patients tolerated one cycle or two cycles (5 of 9 patients) of 6 weekly instillations without significant complications . There was no increased toxicity compared with a control group of 39 patients without reflux who were receiving BCG . Topical intravesical immunotherapy with BCG in patients with vesicorenal reflux can be given without a rise in the complication rate; indeed, in view of the multifocal appearance of urothelial carcinoma in this subgroup, this type of therapy even appears to be desirable. Urol Res, 1989, 17(1), 47 - 55 Immunological aspects of intravesical administration of Bacillus Calmette-Guérin (BCG) in the guinea pig; van der Meijden AP et al.; The only form of effective immunotherapy for human cancer is the intravesical administration of BCG in the treatment of superficial bladder tumors . In this study BCG was administered intravesically (once a week for six consecutive weeks) in guinea pigs to investigate the immune response . Both specific immune responses against BCG itself and nonspecific immune responses such as leukocyte subset distribution, mitogenic (ConA, PHA, LPS) lymphocyte stimulation and spontaneous cytotoxic (NK and LAK cell) activity were determined in the regional draining (iliac) lymph node and in the spleen . The PPD skin test became positive after the fourth intravesical instillation of BCG when 1 X 10(5) culturable particles or higher dosages of BCG were administered . A low dose of 1 X 10(3) c.p . of BCG induced a positive PPD skin reaction in 2 of 4 animals after six BCG instillations . After six BCG instillation BCG immune lymphocytes were present both in the iliac lymph nodes and in the spleen . No differences in mitogenic responses to ConA, PHA and LPS were observed in iliac lymph nodes or the spleen between placebo or BCG treated animals . In the regional iliac lymph nodes a significantly increased number of leukocytes was present . These lymph node cells showed a significant increase in the expression of MHC class II (Ia) immune response antigen on their surface . In the spleen no differences were observed after BCG administration regarding the number of cells present and in terms of the Ia antigen expression of the leukocytes.(ABSTRACT TRUNCATED AT 250 WORDS) Lab Anim Sci, 1989 Jan, 39(1), 16 - 20 Enterohepatitis in Mongolian gerbils (Meriones unguiculatus) inoculated perorally with Tyzzer's organism (Bacillus piliformis); Yokomori K et al.; Enterohepatitis was produced in Mongolian gerbils by intragastric inoculation with Tyzzer's organism from natural infection of a gerbil . Death occurred in 50 to 60% animals 5 to 7 days postinoculation (p.i.) . On day 3 p.i., when a few necrotic foci appeared in the liver, a large amount of bacterial antigen was present within ileocecal enterocytes and reticuloendothelial cells of the Peyer's patches . Neutrophil and monocyte infiltration was found in the lamina propria . On day 5 or 6 p.i . there was severe necrotizing and hemorrhagic ileotyphlocolitis . Bacterial antigen was abundant within not only enterocytes, but also smooth muscle cells of the ileum and jejunum as well as reticular cells of the mesenteric lymph nodes . On day 7 p.i . the intestinal lesions subsided in the presence of fewer bacteria, while necrotizing hepatitis became well developed . The results indicated that Mongolian gerbils were highly susceptible to the oral route of infection with the Tyzzer's organism. J Invertebr Pathol, 1989 Jan, 53(1), 12 - 20 Ingestion, dissolution, and proteolysis of the Bacillus sphaericus toxin by mosquito larvae; Aly C et al.; Larvae of Culex quinquefasciatus are much more susceptible to the toxin of Bacillus sphaericus than are larvae of Aedes aegypti . In the present study, the rate of ingestion, dissolution, and the cleavage by midgut proteases of the B . sphaericus toxin were compared in larvae of these species to determine whether these factors account for the differences in susceptibility . During filter feeding, larvae of both species removed significant quantities of B . sphaericus toxin from suspensions . Filtration rates for 1 hr, the time at which C . quinquefasciatus exhibited marked intoxication, were higher for A . aegypti (576-713 microliters/larva/hr) than for C . quinquefasciatus (446-544 microliters/larva/hr) . Within 24 hr of exposure, A . aegypti larvae ingested 97-99% of the toxin particulates and suffered not more than 10% mortality in suspensions which induced complete mortality in C . quinquefasciatus within 2 hr of exposure . Quantification of the particulate toxin present in larvae after exposure to B . sphaericus suspensions revealed that larvae of both species contained only minor amounts of the toxin, suggesting the larvae had been able to solubilize the toxin after ingestion . Proteases recovered from the feces of larvae cleaved at 43-kDa protein isolated from B . sphaericus toxin extract to 40 kDa in both species . Thus, differences in susceptibility to the B . sphaericus toxin between A . aegypti and C . quinquefasciatus are not due to differences in rates of ingestion, dissolution, or the specificity of proteases. J Bacteriol, 1989 Jan, 171(1), 369 - 74 Pattern of action of Bacillus stearothermophilus neopullulanase on pullulan; Imanaka T et al.; The action of neopullulanase from Bacillus stearothermophilus on many oligosaccharides was tested . The enzyme hydrolyzed not only alpha-(1----4)-glucosidic linkages but also specific alpha-(1----6)-glucosidic linkages of several branched oligosaccharides . When pullulan was used as a substrate, panose, maltose, and glucose, in that order, were produced as final products at a final molar ratio of 3:1:1 . According to these results, we proposed a model for the pattern of action of neopullulanase on pullulan as follows . In the first step, the enzyme hydrolyzes only alpha-(1----4)-glucosidic linkages on the nonreducing side of alpha-(1----6) linkages of pullulan and produces panose and several intermediate products composed of some panose units . In the second step, taking 6(2)-O-alpha-(6(3)-O-alpha-glucosyl-maltotriosyl)-maltose as an example of one of the intermediate products, the enzyme hydrolyzes either alpha-(1----4) (the same position as that described above) or alpha-(1----6) linkages and produces panose or 6(3)-O-alpha-glucosyl-maltotriose plus maltose, respectively . In the third step, the alpha-(1----4) linkage of 6(3)-O-alpha-glucosyl-maltotriose is hydrolyzed by the enzyme, and glucose and another panose are produced . To confirm the model of the pattern of action, we extracted intermediate products produced from pullulan by neopullulanase and analyzed the structures by glucoamylase, pullulanase, and neopullulanase analyses . The experimental results supported the above-mentioned model of the pattern of action of neopullulanase on pullulan. Food Addit Contam, 1989 Jan-Mar, 6(1), 79 - 89 Excretion of penicillins and cephalexin in bovine milk following intramammary administration; Moretain JP et al.; The elimination into bovine milk of beta-lactam antibiotic residues (procaine penicillin G, cloxacillin, ampicillin, oxacillin, cephalexin) following intramammary administration of 10 preparations marketed in France was studied . The quantitative analysis of residues was carried out by a microbiological agar diffusion method using Bacillus stearothermophilus . Sensitivity ranged from 0.001 I.U./ml for procaine penicillin G and 0.001 micrograms/ml for ampicillin to 0.02 micrograms/ml for cephalexin . The mean periods of elimination on which withholding times are based were between four and seven milkings according to the drugs administered. Food Addit Contam, 1989 Jan-Mar, 6(1), 71 - 8 Excretion of tetracycline and chlortetracycline in eggs after oral medication of laying hens; Roudaut B et al.; After dosing laying hens orally with tetracycline (TC) through either drinking water (0.25 and 0.5 g/l for 5 days) or feed (300 and 600 ppm for 7 days), and chlortetracycline (CTC) through feed (600 ppm) residues were determined by an agar plate diffusion technique in cylinders with Bacillus cereus as test-organism, separately for albumen and for yolk . The sensitivity threshold was 0.07 micrograms/g in albumen and 0.15 micrograms/g in yolk for TC and 0.01 micrograms/g in albumen and 0.06 micrograms/g in yolk for CTC . Drug excretion via egg was 3-fold higher for TC than for CTC . The drug was excreted preferentially into the yolk (about 75% of the total amount) and the elimination period lasted between 6 and 11 days for TC and 9 days for CTC, after treatment . Tetracycline use in laying hens is discussed, taking into consideration the proposals presented by the joint FAO/WHO Expert Committee on Food Additives. J Urol, 1989 Jan, 141(1), 22 - 9 Superficial bladder cancer treated with bacillus Calmette-Guerin: a multivariate analysis of factors affecting tumor progression; Herr HW et al.; A multivariate analysis was performed on data from 221 patients with superficial bladder tumors (papilloma in 30, grade II to III stage Ta in 51, grade II to III stage Tis in 111 and grade II to III stage T1 in 29) who were treated with intravesical bacillus Calmette-Guerin and followed for a minimum of 24 months or until progression . The purpose of this analysis was to identify prognostic variables predictive of tumor progression defined as muscle invasion, metastasis or endoscopically uncontrolled superficial bladder carcinoma involving the bladder and/or prostatic urethra . Variables examined before bacillus Calmette-Guerin, and at 3 and 6 months after bacillus Calmette-Guerin included age, sex, race, purified protein derivative reaction, duration of disease, tumor category, tumor grade, multifocality, results of cytology, flow cytometry, cystoscopy, biopsy, prior chemotherapy and bacillus Calmette-Guerin treatment regimen . Significant variables (Cox regression analysis, p less than 0.07) for tumor progression were before bacillus Calmette-Guerin--stage T1 tumors and duration of disease less than 1 year, at 3 months after bacillus Calmette-Guerin--stage T1 tumor, duration of disease less than 1 year, positive cytology studies and multifocality, and at 6 months after bacillus Calmette-Guerin--stage T1 tumor, positive cytology and positive biopsy other than stage T1 tumors . Prognostic risk groups were best defined at 6 months after bacillus Calmette-Guerin, the probability of tumor progression thereafter being at 1, 3 and 5 years, respectively, as follows: for risk group 1 (T1 tumor)--71, 100 and 100 per cent, for risk group 2 (positive biopsy other than T1 plus positive cytology)--25, 79 and 100 per cent, for risk group 3 (either positive biopsy other than stage T1 or positive cytology studies)--18, 40 and greater than 81 per cent, and for risk group 4 (negative biopsy and negative cytology studies)--2, 11 and 26 per cent, respectively . Evaluation of patients with superficial bladder carcinoma at 6 months after intravesical bacillus Calmette-Guerin therapy identifies the probability of tumor progression . Patients at high risk for tumor progression require alternative treatment strategies, whereas low risk patients can be observed for further therapy if necessary. Z Erkr Atmungsorgane, 1989, 173(1), 40 - 5 {Analysis of tuberculosis in foreigners in the GDR}; Weinecke W; In consequence of National Programmes the prevalence of tuberculosis has strongly decreased in the GDR and other industrial states . On the other hand a high prevalence of tuberculosis exists still in most of the developing countries . By the increasing number of immigrants from these high prevalence countries also a higher incidence of tuberculosis cases in this population was observed . The higher risk of infection by bacillary cases must also be taken in consideration . Results of a retrospective analysis of 197 TB-cases in immigrants who were treated in 11 hospitals for lung diseases and tuberculosis in the GDR during the years 1981-1985 are communicated . Thus a contribution for improving case finding, treatment and follow-up of immigrants with tuberculosis in the GDR should be given. Cancer Immunol Immunother, 1989, 28(4), 287 - 95 Immune reactions in patients with superficial bladder cancer after intradermal and intravesical treatment with bacillus Calmette-Guérin; van der Meijden AP et al.; The immune reactivity of patients with strongly recurrent superficial bladder cancer was followed after combined intravesical and intradermal bacillus Calmette-Guerin (BCG) immunotherapy . All patients in this study were previously treated without success with intravesical chemotherapy . The BCG treatment regimen consisted of weekly administrations with BCG (RIVM) for six consecutive weeks, both intravesically and intradermally . In this study, sera and peripheral blood leukocytes (PBL) of patients were tested serially . Besides BCG-antigen-specific reactions, e.g . skin reactivity to purified protein derivatives of Mycobacterium tuberculosis (PPD), antibody formation and antigen stimulation of PBL in vitro, non-antigen-specific immune reactivities were also measured, e.g . mitogen response and spontaneous cytotoxic activity of PBL . In addition the antibody response to bladder carcinoma antigens and the cytotoxic activity of PBL for the bladder carcinoma cell line T24 and the natural-killer-sensitive K562 cell line were investigated . The results obtained from the various assays were evaluated for their prognostic value in relation to the length of the tumor-free interval after the BCG treatment . Because sera and PBL were only obtained during the first 6 months after the BCG treatment, the immune reactivity was compared to the clinical results at that same time . At 6 months after therapy 12 out of 40 BCG-treated patients were tumor-free whereas 28 out of 40 showed a recurrence . Skin reactivity to tuberculin PPD was measured in 40 patients during a period of 3-6 months after therapy . Of patients who showed a recurrence of the tumor within 6 months, 48% of them showed a transient response or developed no response at all to PPD . In the group of patients with a longer tumor-free period (n = 10), only one patient lost the response to tuberculin PPD . Although PBL of a limited number of patients were tested, it was observed that the cytotoxicity to the bladder carcinoma cell line T24, and the natural-killer-sensitive K562 cell line increased in a number of the patients (7 out of 14, and 9 out of 14 respectively) . Reactivity of PBL to mitogens and subset distribution (ratio T-helper: T-suppressor/cytotoxic) were not influenced by the BCG treatment . Antibody response to mycobacterial antigen was detected in 9 out of 23 patients investigated . Of these 9 patients, 8 belonged to the group with a recurrence of the tumor within 6 months (n = 17) . There was no correlation between the skin reactivity and the antibody response to tuberculin PPD.(ABSTRACT TRUNCATED AT 400 WORDS) Ann Med Interne (Paris), 1989, 140(4), 269 - 73 {Patients hospitalized with pulmonary tuberculosis presenting Koch's bacillus in direct sputum examination . Clinical, radiologic and bacteriologic characteristics}; Kuaban C et al.; To determine the influence of socio-economic, clinical and radiological factors on the detection of a Mycobacterium tuberculosis-positive direct sputum smear, we studied 531 patients hospitalized for pleuropulmonary tuberculosis over a 5 year period . M . tuberculosis positivity of the expectorate smear was found more frequently in clinically detected (43%) than in radiologically detected (26%) tuberculosis (P less than 0.001) . On admission, a M . tuberculosis-positive sputum smear was more common in patients under 40 years old (P less than 0.02), blacks (P less than 0.05) and alcoholics (P = 0.001) . M . tuberculosis positivity on direct sputum smears was more often associated with general (asthenia, sweating, fever greater than 38 degrees C) and functional respiratory symptoms (cough and sputum production) (P less than 0.003), bilateral diffusion of lung lesions and/or excavation on chest roentgenography (P less than 0.0001) . Nevertheless, 21% of the asymptomatic radiologically detected tuberculosis patients had positive direct expectorate smears and should be considered contagious. Genet Epidemiol, 1989, 6(4), 501 - 16 Modeling the age-of-onset function in segregation analysis: a causal scheme for leprosy; Abel L et al.; Several methods have been proposed to take into account the variable age of onset of a disease in genetic analysis . A different approach is presented from an etiological point of view . To illustrate the method, we used leprosy, an infectious disease with a variable age of onset depending on both the time of contamination with the bacillus and the latency of the disease; the role of a major gene in the susceptibility to this disease has been recently detected . The age-of-onset function was modeled to account for the two temporal processes: contamination event and incubation period . For genetic analysis, this function was combined with the probability of being susceptible to the disease, which was expressed by the use of regressive models . To test this new approach, ten sets of 500 nuclear families were simulated considering different hypotheses of contamination risks, which were either constant or dependent on contacts with contagious leprosy patients, and varying the extent to which the disease is heritable . Analyses of these data using two versions of the model indicate that the model can detect familial correlations in variable age of onset and discriminate between the different simulated effects. Toxicon, 1989, 27(6), 637 - 45 Release of PI-anchoring enzymes and other effects of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis on TN-368 cells from a moth ovary; Ikezawa H et al.; Release of PI-anchoring enzymes and other effects of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis on TN-368 cells from a moth ovary . Toxicon 27, 637-645, 1989.--The effect of phosphatidylinositol-specific phospholipase C(PIPLC) from Bacillus thuringiensis was investigated on TN-368 cells, derived from the ovary of a moth, Trichoplusia ni . Quantitative analysis of lipids showed that phosphatidylinositol (PI) was contained as one of the major phospholipids in TN-368 cells, whereas sphingomyelin and cholesterol were minor lipid components . When TN-368 cells were treated with PIPLC, significant amounts of alkaline phosphatase, 5'-nucleotidase and beta-glucosidase were released from these cells . Thus, these enzymes were shown to be PI-anchoring proteins in the plasma membrane of these cells . In the presence of 4.2 units of PIPLC, the cell growth of TN-368 was inhibited by 50% . In contrast with normal cells, the cells cultured in the presence of PIPLC became swollen and globular, losing their protoplasmic extensions . Also, there was degeneration of the interior of TN-368 cells cultivated in the presence of PIPLC . Mitochondria became swollen with a decrease in number of granules while the crista turned transparent . Also, an increase in lysosomes was observed and vacuoles seemingly derived from smooth endoplasmic reticula appeared. Eur Urol, 1989, 16(3), 161 - 4 Systemic bacillus Calmette-Guérin infection, 'BCGitis', in patients treated by intravesical bacillus Calmette-Guérin therapy for bladder cancer; Steg A et al.; Among 169 patients treated for supeficial bladder tumor with 150 mg Pasteur-strain bacillus Calmette-Guerin (BCG) intravesical instillation, 5 cases of 'BCGitis' were observed, i.e . a severe systemic BCG infection with bronchopulmonary lesions and granulomatous hepatitis . In 4 cases, the complications appeared at the early stage of treatment (after the 3rd, 6th, 6th and 8th instillations, respectively) . In 1 case, treated with monthly maintenance therapy for 2 years, BCGitis appeared 6 months after treatment had been completed and, in addition to pulmonary basal infiltrate and granulomatous hepatitis, intramedullary granulomatosis was observed . In 3 patients, trauma must be taken into consideration as BCGitis appeared after traumatic instillation with bleeding . All patients were cured by treatment with rifampicin, isoniazid and prednisone. Kokyu To Junkan, 1989 Jan, 37(1), 107 - 11 {A dissecting aortic aneurysm involving a right-sided aortic arch}; Horie K et al.; A 51-year-old woman suddenly developed severe pain in the chest and back, also dyspnea . On admission, she was in a state of preshock . Plain X-ray indicated the lack of the left aortic arch and poor pneumatization in the whole right lung . The thoracic fluid was transparent with yellowish tinge and was contaminated with neither any bacterium nor tubercle bacillus . The response to the Rivalta's reaction was negative . The possibility of pleurisy was, therefore, denied . The ECG and blood biochemical data on the second day suggested the possibility of myocardial ischemia . Plain chest X-ray on the fourth day revealed an increased right pulmonary pneumatization and an enlarged mediastinal shadow toward the aortic arch . Upper pulmonary CT showed a mass on the right side . Enhanced CT disclosed a dissepiment in the center, which was high medially and somewhat low laterally . It was diagnosed as a false lumen due to the lateral displacement of the right aortic arch . Hepatic CT disclosed the tapering of the abdominal aorta from right to left in the prevertebral region . These findings indicated that the aorta descended from the right aortic arch along the right side of the spine and crossed the spine dextrosinistrally at the hepatic level . In addition, dissociant aneurysm was observed in the right aortic arch . Echocardiography showed no evidence of dissociant aneurysm at the aortic base . Chest X-ray, CT and echocardiography showed the dissociation of the aorta from the aortic arch to the abdominal aorta . Thus the diagnosis of De Bakey type III was established . Clinically, DIC and multiorgan disorders were manifested but after medical treatments, the clinical course was uneventful.(ABSTRACT TRUNCATED AT 250 WORDS) Antibiot Khimioter, 1989 Jan, 34(1), 10 - 3 {Relation of bacitracin synthesis and manganese content in Bacillus licheniformis: antibiotic production in colonies forming on media with high levels of Mn2+}; Kuzovnikova TA et al.; Population of B . licheniformis is not homogeneous with respect to its sensitivity to Mn2+ . Active bacitracin producing cultures with high and low sensitivity to exogenic manganese are described . In media containing high sublethal concentrations of Mn2+ there were detected two types of bacitracin-producing colonies i.e . light and dark differing likely in the mechanisms of their cell protection from the metal excess . Since bacitracin production did not always correlate with sensitivity to Mn2+ at the level of the colony formation the use of Mn2+ as a selective factor for isolating inactive strains of B . licheniformis is of low efficiency. Pharm Acta Helv, 1989, 64(3), 68 - 70 {Sterilization of non-air evacuated infusion flasks containing little water}; Kownacki E et al.; Studies have been carried out in connection with the sterilization of glass infusion bottles from which air had not been evacuated and containing certain limited quantities of water . Bottles with a capacity of 500 and 1000 cm3 were filled with 1-3 cm3 distilled water and sterilized in an autoclave for 20 min . set at a temperature of 121 degrees C reaching up to 126 degrees C . The sterility was tested by introducing bags containing Bacillus stearothermophilus spores into the centre of the bottles . Following sterilization the bottles were examined for the growth of spores allowing an incubation period of 7 days at 37 degrees C . Based on the actual conditions of this investigation, it would appear that a bottle from which air has not been evacuated should contain at least 2% (V/V) distilled water, when sterilized in an autoclave . It is, however, strongly recommended that the present findings should be validated based on actual conditions prevailing for individual sterilization procedures. Gematol Transfuziol, 1989 Jan, 34(1), 11 - 4 {Bacteriotherapy of intestinal dysbacteriosis in patients with acute leukemia}; Tolkacheva TV et al.; The results have been presented of correcting intestinal microflora in 127 patients with acute leukemia by preparations containing Bacillus bifidus and acidophilus: dried and milk bifidum-bacterin, biofructolact, acidophilic milk. Eur Urol, 1989, 16(1), 28 - 30 Immunoprophylaxis with Calmette-Guérin bacillus in recurrent superficial bladder tumors; Llopis B et al.; 26 patients with superficial bladder cancer underwent complete removal of tumors by transurethral resection and entered a prospective trial of immunoprophylaxis with Calmette-Guerin bacillus (BCG) . The entire group had recurrent tumors that failed to respond to previous prophylaxis with thiotepa (13 patients) or other drugs (13 patients) . Intravesical administration of BCG reduced the recurrence index and increased the disease-free interval . Side effects were mild and well tolerated . BCG seems to be effective in the prophylaxis of superficial bladder tumor recurrence after the failure of other drugs. Med Parazitol (Mosk), 1989 Jan-Feb, (1), 81 - 3 {Ineffectiveness of bacterial insecticides against egg laying by female mosquitoes}; Voitsik AA et al.; Insecticides on the base of Bacillus thuringiensis and B . sphaericus proved to be ineffective against laying of eggs by female mosquitoes in the water containing these formulations. Med Pediatr Oncol, 1989, 17(2), 155 - 8 Very late recurrence of childhood acute lymphoblastic leukemia treated with chemoimmunotherapy: a report of three cases occurring 19, 11, and 9 years after discontinuation of chemotherapy; Salloum E et al.; Current therapeutic modalities for childhood acute lymphoblastic leukemia (ALL) are associated with a high cure rate, and recurrences more than 4 years after therapy cessation are very unusual . We report three cases of exceptionally late recurrences of childhood ALL after cessation of chemotherapy (CT) given for respective periods of 8, 7, and 24 months . CT was followed by maintenance immunotherapy (IMT) with Bacillus Calmette-Guerin (BCG) and allogeneic leukemic lymphoblasts pretreated with formaldehyde or irradiated in vitro . Leukemic recurrences were observed 19, 11, and 9 years after cessation of CT and appeared morphologically similar to the original blasts . A second complete remission was easily achieved in all three patients, but two went on to repeated relapse (one has died following the fourth recurrence) . We speculate that some residual leukemic cells, remaining after the inadequate, short-term CT, were responsible for these unusual evolutions, and we question a possible delaying role of IMT in prolonging remission . Other possible etiologies are discussed. Cancer Immunol Immunother, 1989, 28(4), 282 - 6 The significance of conversion of skin reactivity to efficacy of bacillus Calmette-Guérin (BCG) vaccinations given immediately after radical surgery in stage II melanoma patients; Cascinelli N et al.; A group of 668 stage II melanoma patients was entered into a randomized prospective study aimed at evaluating the efficacy of adjuvant BCG, 5-(dimethyltriazeno)imidazole-4-carboxamide (DTIC), or a combination of the two, given immediately after radical lymph node dissection . Of these, 176 patients received BCG and 164 BCG plus DTIC . These 340 patients had histologically proven metastatic nodes and 156 had a negative skin reactivity to BCG at the beginning of treatment . The distribution of known prognostic factors (sex, age, number of positive nodes, extracapsular invasion) was balanced in the groups of patients either with initially negative or with positive skin reactivity . All patients who were initially non-reactive to BCG developed skin reactivity after 6.7 +/- 9 BCG vaccinations . Disease-free and overall survival of patients receiving BCG or BCG + DTIC with an initially negative skin reactivity to BCG was significantly (P = 7 x 10(-3) better than that observed in patients with an initial positive skin reactivity . This finding was still evident after adjustment for other known prognostic criteria (P = 0.02) . It seems likely that the initial BCG skin reactivity as such marks the prognosis; however, some therapeutic effect of BCG treatment in patients having initially no skin reactivity to BCG, can not be ruled out. Eur J Cancer Clin Oncol, 1989, 25 Suppl 3, S31 - 6 Malignant melanoma--prognosis and actual treatment strategies with chemotherapy and biological response modifiers; Bergmann L; The prognosis of malignant melanoma (MM) depends on the level of invasion, vertical tumour size, location of the primary, clinical stage, and sex . Whereas MMs are potentially curable in the early stage of disease, the therapeutic possibilities are very limited in advanced and disseminated MM . Most chemotherapeutic agents lack sufficient activity in MM especially with regard to survival . Dacarbazine (DTIC) is the most effective drug in MM with response rates of 20-25% followed by other drugs such as melphalan with 15-20%, hydroxyurea and platin derivates . Multidrug regimens were not shown to be more effective than DTIC alone . Radiotherapy may be relevant in local treatment of metastases . With regard to the poor prognosis and limited therapeutic approaches in advanced and disseminated MM, new strategies are required . In this context immunotherapeutic strategies with biological response modifiers are of interest for adjuvant or palliative approaches . Earlier trials with Bacillus Calmette-Guerin (BCG) +/- DTIC as adjuvant or palliative treatment revealed no effect of BCG on the prognosis . Alpha-interferon (alpha-IFN) was shown to induce remissions in about 15% and gamma-IFN in about 10% of patients . A very interesting new approach is the induction and/or activation of autologous cytotoxic cells by systemic administration of recombinant interleukin-2 (rIL-2) with response rates of 20-25% and the in vivo propagation and transfer of so-called tumour infiltrating lymphocytes . Further trials combining rIL-2 with other cytokines, chemotherapy, tumour vaccination or monoclonals against melanoma cells are required. Acta Leprol, 1989, 7(1), 37 - 40 {Immunologic aspects of leprosy}; Ayed K; Lepromatous leprosy is accompanied by a complex deficit in cell mediated immunity concerning Mycobacterium leprae . The physiological mechanism of this deficit remains unknown . According to some studies there may be, in those suffering from leprosy, a trouble in the macrophage and presentation of antigens . Other studies suggest an increase in the suppressive activity of lymphocytes . Recently some authors emphasized the deficit in the production of interleukin 2 . It is difficult, for the time being, to find out whether such immunological abnormalities are primitive or secondary to the accumulation of the bacillus into the organism. Urol Res, 1989, 17(5), 299 - 303 Analysis of mucosal bladder leucocyte subpopulations in patients treated with intravesical Bacillus Calmette-Guerin; Peuchmaur M et al.; Immunohistochemical techniques were used to investigate leucocyte subpopulations in the bladders of patients with superficial transitional cell carcinoma treated with BCG Pasteur . Leucocyte subsets were enumerated with a panel of monoclonal antibodies which included CD3, CD4, CD8, TQ1, Leu7, CD15, HLA-DR, CD25, CD22 . We demonstrated in the bladders of patients treated with BCG a particular lymphocyte population; the major subset was an inducer (CD4+, TQ1-) which was activated (CD25+, HLA-DR+) and associated with polymorphonuclear eosinophils . There was neither inducer of suppression nor major cytotoxic/suppressive subsets . CD8+ and NK cells could not be the primary mediators of BCG activity . These data supported the hypothesis of a helper T lymphocyte activity associated with lymphokine production and activation of effector killer cells. Z Erkr Atmungsorgane, 1989, 173(1), 18 - 22 {Position, development and trends in the epidemiology of tuberculosis in the GDR 1982-1987}; Schilling W et al.; On the occasion of the commemoration of the 100th anniversary of the discovery of the tubercle bacillus by Robert Koch, in 1982, it could be stated that in GDR, like in other developed countries, tuberculosis is not any more a serious problem of public health but has changed to a much reduced endemic disease . After evaluation of the most important parameters of the tuberculosis problem, morbidity, mortality, and infection rate, conclusions are drawn for the future strategy of tuberculosis control . After the rapid decline of the incidence of new tuberculosis cases during the seventies, also during the last 5 years the epidemiology figures were favourable . But it has to be stated, that during the last years the decrease of the incidence of new cases of all forms of the disease was less pronounced, and with bacteriologically confirmed pulmonary tuberculosis a stagnation of the incidence occurred in the last four-years-period . Today, under our circumstances, tuberculosis is mainly a disease of middle and higher age . Therefore, the strategy of detection of new cases has to be adapted to and developed according to these conditions . New methods of case finding, detection of infection sources, screening or risk groups, and the investigation of patients with bronchopulmonary symptoms must be developed . Also during the next years, strict surveillance of tuberculosis remains mandatory. Carlsberg Res Commun, 1989, 54(2), 41 - 54 Hybrid bacillus endo-(1-3,1-4)-beta-glucanases: construction of recombinant genes and molecular properties of the gene products; Borriss R et al.; Hybrid beta-glucanase genes were constructed by the reciprocal exchange of the two halves of the isolated beta-glucanase genes from Bacillus amyloliquefaciens and B . macerans . The beta-glucanase hybrid enzyme 1 (H1) contains the 107 amino-terminal residues of mature B . amyloliquefaciens beta-glucanase and the 107 carboxyl-terminal amino acid residues of B . macerans beta-glucanase . The reciprocal beta-glucanase hybrid enzyme 2 (H2) consists of the 105 amino-terminal residues from the B . macerans enzyme and the carboxyl-terminal 107 amino acids from B . amyloliquefaciens . The biochemical properties of the two hybrid enzymes differ significantly from each other as well as from both parental beta-glucanases . Hybrid beta-glucanase H1 exhibits increased thermostability in comparison to other beta-glucanases, especially in an acidic environment . This hybrid enzyme has maximum activity between pH 5.6 and 6.6, whereas the pH-optimum for enzymatic activity of B . amyloliquefaciens beta-glucanase was found to be at pH 6 to 7 and for B . macerans at pH 6.0 to 7.5 . Hybrid enzyme 1 being more heat stable than both parental enzymes represents a case of intragenic heterosis . Hybrid beta-glucanase 2 (H2) was found to be more thermolabile than the naturally occurring beta-glucanases it was derived from and the pH-optimum for enzymatic activity was determined to be between pH 7 and pH 8. Arch Microbiol, 1989, 152(3), 280 - 8 Methanol metabolism in thermotolerant methylotrophic Bacillus strains involving a novel catabolic NAD-dependent methanol dehydrogenase as a key enzyme; Arfman N et al.; The enzymology of methanol utilization in thermotolerant methylotrophic Bacillus strains was investigated . In all strains an immunologically related NAD-dependent methanol dehydrogenase was involved in the initial oxidation of methanol . In cells of Bacillus sp . C1 grown under methanol-limiting conditions this enzyme constituted a high percentage of total soluble protein . The methanol dehydrogenase from this organism was purified to homogeneity and characterized . In cell-free extracts the enzyme displayed biphasic kinetics towards methanol, with apparent Km values of 3.8 and 166 mM . Carbon assimilation was by way of the fructose-1,6-bisphosphate aldolase cleavage and transketolase/transaldolase rearrangement variant of the RuMP cycle of formaldehyde fixation . The key enzymes of the RuMP cycle, hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), were present at very high levels of activity . Failure of whole cells to oxidize formate, and the absence of formaldehyde- and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formaldehyde via HPS . A comparison of the levels of methanol dehydrogenase and HPS in cells of Bacillus sp . C1 grown on methanol and glucose suggested that the synthesis of these enzymes is not under coordinate control. Prog Clin Biol Res, 1989, 310, 325 - 34 Systemic bacillus Calmette-Guerin infection in patients treated by intravesical BCG therapy for superficial bladder cancer; Steg A et al.; Among 169 patients treated for superficial bladder tumors with intravesical instillations of 150 mg BCG-Pasteur, five developed BCG-itis--a severe systemic infection with bronchopulmonary lesions and granulomatous hepatitis . In four cases, the complication appeared early during treatment (after three, six, six and eight instillations respectively) . In one case, BCG-itis appeared 6 months after completion of 2 years monthly maintenance therapy . In addition to pulmonary basal infiltration and granulomatous hepatitis, intramedullary granulomatosis was observed . In three patients, the role of trauma has to be considered, as BCG-itis appeared after a traumatic instillation with bleeding . All patients were cured by combined treatment with Rifampicin, Isoniazid and Prednisolone. Nauchnye Doki Vyss Shkoly Biol Nauki, 1989, (5), 85 - 9 {Bacillus thuringiensis fluorescence induced by anaerobiosis and its relation to the cellular functional state}; Efimtsev EI et al.; The fluorescence spectra of Bacillus thuringiensis native cells at 340 nm induction have been investigated . The spectral curve has 440-445 and 510-515 nm maximums of fluorescence of pyridine nucleotides (PN) and flavoproteins (FP) . The induction of PN and FP fluorescence under anaerobiosis resulted from bacterium breath has been found . The fluorescence induction is connected with the viability of cells . The amplitude of induced fluorescence is in a linear dependence on the number of vegetative cells in medium . The given experimental approaches may be used in a course of development of methods of continuous control over the physiological state change of microorganisms. Arch Microbiol, 1989, 152(1), 58 - 63 Replacement of potassium ions by ammonium ions in different micro-organisms grown in potassium-limited chemostat culture; Buurman ET et al.; The biomass concentration extant in potassium-limited cultures of either Klebsiella pneumoniae or Bacillus stearothermophilus (when growing at a fixed temperature and dilution rate in a glucose/ammonium salts medium) increased progressively as the medium pH value was raised step-wise from 7.0 to 8.5 . Because the macromolecular composition of the organisms did not vary significantly, this increase in biomass could not be attributed to an accumulation of storage-type polymers but appeared to reflect a pH-dependent decrease in the cells' minimum K+ requirement . Significantly, this effect of pH was not evident with cultures in which no ammonium salts were present and in which either glutamate or nitrate was added as the sole nitrogen source; however, it was again manifest when various concentrations of NH4Cl were added to the glutamate-containing medium . This suggested a functional replacement of K+ by NH4+, a proposition consistent with the close similarity of the ionic radii of the potassium ion (1.33 A) and the ammonium ion (1.43 A) . At pH 8.0, and with a medium containing both glutamate (30 mM) and NH4Cl (100 mM), cultures of B . stearothermophilus would grow without added potassium at a maximum rate of 0.7 h-1 . Under these conditions the cells contained maximally 0.1% (w/w) potassium (derived from contaminating amounts of this element in the medium constituents), a value which should be compared with one of 1.4% (w/w) for cells growing in a potassium-limited medium containing initially 0.5 mM K+.(ABSTRACT TRUNCATED AT 250 WORDS) Arch Microbiol, 1989, 151(2), 91 - 4 The nucleotide sequence of the lipo-penicillinase gene of alkalophilic Bacillus sp . strain 170; Kato C et al.; The lipo-penicillinase (LIPEN) gene from an alkalophilic Bacillus sp . strain 170 was cloned in Escherichia coli using the vector pHSG399 . A plasmid, pFAP121, was isolated from an ampicillin resistant transformant and the cloned LIPEN gene was found to be in a 2.2 kb DNA fragment . The nucleotide sequence of a 1.9 kb segment encoding the LIPEN was determined . This segment showed an open reading frame which would encode a polypeptide of 310 amino acids . The amino acid sequence of this LIPEN gene product has strong homology with those of the Bacillus cereus beta-lactamase III and Bacillus licheniformis penicillinase. Pediatr Med Chir, 1989 Jan-Feb, 11(1), 1 - 11 {Tuberculosis in children in the early 90's}; Battistini A et al.; The natural history of tuberculosis was re-examined, and especially the relation between contagion and first infection on one side and, on the other, primary tuberculosis (in children) and secondary tuberculosis in adult . With regard to the primary tuberculosis, the different clinical pictures were taken into consideration and it was reported how symptoms change as years go by (see miliary tuberculosis) . As for the diagnostic aspect the importance of a quantitative interpretation of Mantoux reaction, was emphasized . In the final judgement, all factors that may change the results can therefore by considered: from the intercurrent viral infection to the patient's age, to the spreading of atypical mycobacteria in that area . The screening performed with the multi-injections technique is necessary to control the diffusion of the infection in the general population . In Italy, 1-2% of children aged 6 to 14 years old, are skin positive but the percentage varies widely in the different areas . However, it was higher than that observed in other European countries and far from the 1% incidence for 14 year olds, as hoped by the World Health Organization . After re-examining the characteristics of the four most important anti-tubercular drugs with particular attention to possible hepatotoxic effects of the isoniazid-rifampicin association, the authors analyses new therapeutic strategies, especially the one that tends to shorten treatment periods . This approach is effective in adults but has not yet been definitively confirmed in children . The surgical approach is more standardized in superficial adenitis where extirpation is indicated in atypical mycobacteria infections rather than in those due to Koch's bacillus . As far as indications of chemoprophylaxis are concerned it would seem that an international arrangement has now been reached that on the whole extends the indications to this type of treatment . The only uncertainty is whether adults, with intradermal reactions to tuberculin should undergo chemoprophylaxis if the time of contagion is unknown and the subject does not show any particular risk factors . On the grounds of data from literature, the results of vaccination are contrasting . Many factors could be responsible for this, especially the use, till a few years ago, of a non-standardized vaccine . However, vaccination remains an efficient method in populations at risk, listed in Italian law too . Nevertheless, according to the law, the subjects who would most benefit by vaccination are excluded (children under the age of 5 years and over the age of 15 years) . However vaccination is getting less important in the general population.(ABSTRACT TRUNCATED AT 400 WORDS) Diagn Microbiol Infect Dis, 1989 Jan-Feb, 12(1), 93 - 9 Cefotaxime and desacetylcefotaxime in neonates and children: a review of microbiologic, pharmacokinetic, and clinical experience; Jacobs RF et al.; Over the past 5 yr, we have conducted two clinical and two pharmacokinetic investigations of cefotaxime (CTX) and desacetylcefotaxime (dCTX) in neonates, infants, and children . A total of 50 children with culture-proven bacterial meningitis were randomized to receive either 200 mg/kg/day of CTX (n = 23, mean age 24.4 mo) or standard doses of ampicillin (AMP) and chloramphenicol succinate (CAPS; n = 27, mean age 16.6 mo) . Results were similar between the CTX and Amp/CAPS groups for clinical/microbiological cures (100% versus 96%, respectively) and for survival without sequelae (78% vs . 77%, respectively) . All bacterial isolates were sensitive to CTX, and the comparison of the MIC/MBC values for CTX to the CSF bactericidal titers suggested antimicrobial activity for dCTX . In a second clinical trial, 20 infants (1 wk-3 mo) were treated with 200 mg/kg/day of CTX for Gram-negative enteric bacillary meningitis . Cultures of CSF obtained 24 hr after the initiation of treatment were sterile in all subjects . Survival and complication rates of 95% and 21%, respectively, were observed . This compared favorably to previously published experiences with alternate treatment regimens for Gram-negative meningitis in the newborn . In both meningitis studies, the safety profile for CTX was excellent.(ABSTRACT TRUNCATED AT 250 WORDS) Ethiop Med J, 1989 Jan, 27(1), 15 - 20 Plasmid DNA analysis by agarose gel electrophoresis on an epidemic-associated strain of trimethoprim resistant Shiga's bacillus from Gimira Wereda (Keffa administrative region, southwest Ethiopia); Gebre-Yohannes A et al.; A combination of genetic mating experiments and plasmid DNA analysis by gel electrophoresis was carried out on an epidemic-associated Shiga's bacillus with resistance type ACSSuTTp, isolated in 1983 from Gimira Wereda, Keffa Administrative Region, South West Ethiopia . The donor strain contained 9 plasmid species . Their sizes in megadaltons (Mdal) were: 120, 40, 35, 12.5, 6.0, 5.1, 4.3, 2.5 and 1.7 . Analysis of Escherichia coli K12 transconjugants revealed that the 40, 35, and 4.3 Mdal plasmids coded for tetracyline-trimethoprim, ampicillin -chloramphenicol and streptomycin-sulphadiazine resistance respectively. J Bacteriol, 1989 Jan, 171(1), 521 - 30 A 20-kilodalton protein is required for efficient production of the Bacillus thuringiensis subsp . israelensis 27-kilodalton crystal protein in Escherichia coli; Adams LF et al.; The 27-kilodalton (kDa) mosquitocidal protein gene from Bacillus thuringiensis subsp . israelensis has been cloned as a 10-kilobase (kb) HindIII fragment from plasmid DNA; efficient expression in Escherichia coli KM1 depends on a region of DNA located approximately 4 kb upstream (K . McLean and H . R . Whiteley, J . Bacteriol . 169:1017-1023, 1987) . We have cloned the upstream DNA region and show that it contains a complete open reading frame (ORF) encoding a protein with a molecular mass of 19,584 Da . Sequencing of adjacent stretches of DNA revealed two partial ORFs: one has 55.2% identity in an overlap of 319 amino acids to the putative transposase of IS231 of B . thuringiensis subsp . thuringiensis, and the other, a 78-codon partial ORF, may be the carboxyl terminus of the 67-kDa protein previously observed in maxicells of strain KM1 . A 0.8-kb fragment containing only the 20-kDa protein gene greatly enhanced the expression of the 27-kDa protein in E . coli . The introduction of nonsense codons into the 20-kDa protein gene ORF abolished this effect, indicating that the gene product, not the mRNA or DNA, is required for the enhancement . The effect of the 20-kDa protein gene on various fusions of lacZ to the 27-kDa protein gene suggests that the 20-kDa protein acts after the initiation of translation of the 27-kDa protein gene. Wiad Parazytol, 1989, 35(6), 577 - 83 {A redescription of Capillaria carbonis (Rudolphi, 1819), (Capillariidae), based on material collected from the black cormorant (Phalacrocorax carbo)}; Okulewicz A; A redescription of a nematodes Capillaria carbonis was made on the basis of specimens found for the first time in Phalacrocorax carbo in Poland in Milicz district . It has been found that: the body of male terminates with two processes with a single large papilla, in its distal part surrounded by cuticular pseudobursa formed from four cuticular lobes; spicular sheath non spiny has a transversely-oblique striation; two lateral bacillary bands extend along whole body; number of stichocytes equals 38-42 and 42-45 in males and females respectively; vulva of gravid females appears sometimes with vulvar appendage. Acta Microbiol Hung, 1989, 36(2-3), 119 - 24 Biological and immunological properties of avirulent strain of Listeria innocua; Mencikova E et al.; The strain isolated by Dr . J . H . Welshimer from plants has antigenic formula V (VI) IX; XV; XI; AB, C--serovar 6a, is non-haemolytic, produces lipase, and toxic factor Ei, is avirulent for adult mice, but causes encephalitis in sucklings . In organs of intravenously injected mice the strain persists and multiplies for 1-3 weeks . The protective effect against listerial infections in mice of this strain administered 2-14 days before challenge is dose depending . After 3 weeks induces resistance of guinea pigs to infection with Mycobacterium tuberculosis H37Rv measured by spleen weight and Feldman index . The hypersensitivity induced in animals is detectable by factor Ei and PPD or OT tuberculins using MIF method . A suspension of living cells of this strain injected intraperitoneally causes resistance to Mycobacterium kansasii in mice, measured by inhibition of loss of weight and decrease of the number of bacillus in their lungs. Medicina (B Aires), 1989, 49(1), 48 - 52 {Pleural effusion: diagnostic usefulness of pleural puncture biopsy}; Ferreyra RA et al.; To assess the diagnostic usefulness of thoracocentesis with pleural needle biopsies, we retrospectively studied 316 procedures performed in 254 patients between 1977 and 1984 . Of these, 130 were ultimately found to have pleural malignant disease, with a diagnostic cytologic study in 60% of the patients, a positive pleural biopsy in 52.30% and both methods combined in 81.53% of the patients . The marginal gains from pleural biopsy in the presence of negative cytology results were 63.46% . In 59 patients, the primary neoplasm was lung cancer, the most frequent tumor (45.30%); in second place, 30 patients presented carcinoma of the breast (23%) . Needle biopsy of the pleura proved to be nonspecific for the diagnosis of nonmalignant diseases except for tuberculous pleurisy: in our study 55 patients presented tuberculosis (21.65%) and in 35 of them, the pleural biopsy was characteristic . The culture of pleural fluid revealed Koch bacillus in 5.45% of these patients . Routine culture of biopsy specimen for tubercle bacillus was not carried out . Complications occurred in 4.4% of needle biopsies, with no death. Yi Chuan Xue Bao, 1989, 16(3), 206 - 12 {Study on Bacillus pumilus as a recipient strain for genetic engineering of Bacillus}; Chen QM et al.; Bacillus pumilus 289 can be transformed easily by plasmid pUB110 through protoplast transformation with the frequency of 10(-5)--10(-3), similar to B . subtilis AS 1.1176, a derivative strain of B . subtilis 168 . The regeneration frequency of its protoplast is only slightly lower than B . subtilis AS1.1176 (0.3-12.0% compare to 1.53--24.16%) . Plasmid pUB110 can be maintained in both bacterial strains stably . The frequency of loss of the plasmid in both strains is lower than 3% after 45 generations in LB medium . But it is quite different that the hybrid plasmid (pUB110 with 3.9 kb foreign DNA fragment) can be maintained much more stably in B . pumilus 289 than in B . subtilis AS1.1176 . The frequency of loss of the plasmid is lower than 5% in B . pumilus 289 and 24% in B . subtilis AS1.1176 after 25 generations when they grown in SH medium . The expression level of foreign gene in B . pumilus 289 is also much higher than that in B . subtilis AS1.1176 . Therefore B . pumilus 289 is valuable to be exploited as recipient strain for genetic engineering of Bacillus in the future. Vestn Dermatol Venerol, 1989, (11), 41 - 5 {Secondary candidal infection in patients with chronic dermatoses}; Dovzhanskii SI et al.; Clinical and laboratory examinations of 250 patients with chronic dermatoses (eczema, neurodermatitis, psoriasis, pemphigus, etc.) have revealed secondary candidal involvement of the skin and mucosa in 121 (46.6%) . C . albicans, often associated with bacillary (33.3%) or bacterial (7.3%) microflora, predominated among the yeast-like fungi isolated from the patients . These data point to the necessity of prescribing, besides the antimycotic agents, a complex of pathogenetic therapy including immunomodulators, in order to cut down the terms of treatment and prevent secondary candidal infection in the patients with chronic dermatoses. Arch Virol, 1989, 109(3-4), 207 - 22 Characterization of viral proteins of Oryctes baculovirus and comparison between two geographical isolates; Mohan KS et al.; Bacilliform Oryctes baculovirus particles have been visualized in electron micrographs of midgut sections from virus infected Oryctes rhinoceros beetles . Morphologically the Indian isolate (Oryctes baculovirus, KI) resembled the previously reported Oryctes baculovirus, isolate PV505 . The constituent proteins of baculovirus KI have been analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blots using polyclonal antibodies raised against the complete viral particles, as probes . A total of forty eight viral proteins have been identified . Fourteen viral proteins were located on the viral envelope . Among the proteins constituting the nucleocapsid, three were located internally within the capsid . A 23.5 kDa protein was tightly associated with viral DNA in the nucleocapsid core . Two envelope and seven capsid proteins of KI and PV505 revealed differences in SDS-PAGE profiles and glycosylation patterns . Immunoblotting of KI and PV505 proteins with anti KI antiserum demonstrated antigenic differences between the two viral isolates. Int J Oral Maxillofac Implants, 1989 Summer, 4(2), 139 - 46 Dynamic sterilization of titanium implants with ultraviolet light; Singh S et al.; All implantable devices must be sterile . However, autoclaves produce poor surface properties that jeopardize the integration process . The application of a modified ultraviolet light source has proven to enhance bioreactivity by controlling surface properties, but it lacks validation of its sterilization capabilities . Forty-eight titanium implants were contaminated with spores of the biological indicator Bacillus stearothermophilus and subjected to "dynamic sterilization" by ultraviolet light . Forty-seven of the implants were successfully sterilized, as indicated by not producing turbidity in a suitable growth medium . This sterilization technique only requires a 20-second exposure to achieve sterility. Eur Urol, 1989, 16(6), 401 - 4 Bladder tumors invading the lamina propria (stage A/T1): influence of endovesical bacillus Calmette-Guérin therapy on recurrence and progression; Boccon-Gibod L et al.; 47 patients with transitional cell bladder carcinoma invading the lamina propria (stage A/T1) were treated from 1984 to 1986 by complete transurethral resection followed by 1-3 cycles of endovesical bacillus Calmette-Guerin instillations, and followed 14-64 months by cytology, endoscopy and bladder biopsies, 64% achieved a complete response, 36% recurred (recurrence rate/100 months/patient 2.2), 21% progressed to muscle invasion . Duration of treatment, tumor size or type (solid vs . papillary), presence of carcinoma in situ bore no relation to the final result . The preceding history of T1 bladder tumor appeared associated with a higher risk of progression although not reaching statistical significance . The results were compared to those obtained by transurethral resection in a similar group of 50 patients treated from 1982 to 1984 and followed up 12-100 months, 90% recurred and 34% progressed to muscle invasion with a recurrence rate/100 months/patient of 9.22 . Keeping in mind the limits of a nonrandomized historical comparison, it appears that endovesical bacillus Calmette-Guerin therapy alters favorably the recurrence pattern of T1 bladder cancer. Chin J Biotechnol, 1989, 5(1), 33 - 7 Restriction map of large plasmid pFW1 in Bacillus sphaericus strain 10; Wei BY et al.; Some strains of Bacillus sphaericus have been found to be toxic to mosquito larvae (1) . They differ from Bacillus thuringiensis var . israelensis (BTI) (2,3) in the following aspects: BTI is relatively ineffective in polluted water, and its residual activity in most habitats is limited to a few days after treatment, whereas BS isolates are effective in such habitats, particularly against species of Culex; BS also remains effective longer, as a result of either persistence or recycling . B . sphaericus strain 10 (BS10) was isolated by the Lixiahe Institute of Agricultural Sciences in Jiangsu Province . Its larvicidal activity is much higher than that of BS1593, which was recommended by the World Health Organization for widespread use as a standard strain (1) . The mosquito-larvicide gene of BS1593 has been cloned and expressed in E . coli and B . subtilis (4, 5) . In the present study, BS10 was investigated to establish whether it contains the plasmid and whether homology exists between the mosquito-larvicide gene in BS and that in BTI. Cancer Treat Res, 1989, 46, 195 - 211 Mechanisms of action of intravesical bacillus Calmette-Guerin for bladder cancer; Mikkelsen DJ et al.; Though superficial bladder-cancer patients have been treated with intravesical BCG since 1976, the mechanisms of action remain unknown . Evidence points towards an immune mechanism of BCG-induced antitumor activity . Although specific antitumor immunity may play a role in BCG immunotherapy, additional work is needed to more clearly define this possibility . Several investigators have noted a clinical association between favorable response to BCG therapy and a systemic response (DTH) to BCG antigen, indicated by conversion to a positive purified protein derivative (PPD) skin test . Additional animal studies have, in turn, documented evidence of specific binding between BCG and fibronectin found in the urothelial basement membrane, which appears necessary for the development of both the DTH response and antitumor activity . Other studies have suggested which cellular components of the immune system and lymphokines may be involved in the antitumor response . Further work will be needed to better understand BCG mechanisms, as these findings may be important to other forms of cancer therapy . Questions of immune suppression, genetic influence on the immune response, and immunocompetence at the time of surgery may assume increasing importance . A strong effort will therefore be needed to better understand these mechanisms so that patients may be selected, treated, and followed in a more effective manner. Acta Virol, 1989 Jan, 33(1), 68 - 74 Effects of pH and ionic strength on precipitation of phytopathogenic viruses by polyethylene glycol; Pastorek J et al.; The effects of ionic strength of the solution (changed by varying NaCl concentrations or buffer molarity) on the precipitation with polyethylene glycol (PEG) 6000 were studied on phytopathogenic viruses of different morphology: the isometric red clover mottle virus (RCMV), rod-shaped tobacco mosaic virus, flexuous potato virus X (PVX) and bacilliform alfalfa mosaic virus . With increasing NaCl concentration or buffer molarity up to a certain level (0.1 mol/l), the efficiency of PEG precipitation increased . This relationship did not apply to PVX . The effects of pH on PEG precipitation were studied on RCMV . The efficiency of precipitation increased with decreasing difference between pH of the solution and pI of the virus. Chin J Biotechnol, 1989, 5(1), 1 - 10 Cloning and expression of delta-endotoxin gene of Bacillus thuringiensis in Escherichia coli; Tian YC et al.; Fragments (larger than 4 kb) of partially Sau 3AI-digested plasmid DNA from Bacillus thuringiensis subsp . kenyae 7404 and kurstaki HD-1 were cloned into the BamH I site of pBR322 . Four transformants, containing the corresponding delta-endotoxin gene and producing proteins that reacted with antiserum against crystalline protein, were selected on the basis of results from in situ colony hybridization, radioimmunologic screening, and Western blot analysis . Three of those tested, i.e., the lysates of one transformant (TK89) carrying the delta-endotoxin gene of B.t . 7404 and two transformants (TH12 and TH48) carrying the delta-endotoxin gene of HD-1, were toxic for tobacco budworm (Heliothis assulta) and armyworm (Leucania separata Walker) caterpillars . This is the first report on cloning of the delta-endotoxin gene from B.t . subsp . kenyae, which is different in serotype from the well-studied subsp . kurstaki. Acta Biochim Biophys Hung, 1989, 24(3), 283 - 90 The role of water in the radiation sensitization of acetone; Gazso L et al.; These experiments were aimed at the better understanding of the mechanism of the enhancement of radiation damage brought about by acetone in spores of Bacillus megaterium . Particularly, the intention was to examine the extent of involvement of water content in this action of acetone . The radiosensitization of acetone increased with increasing oxygen concentration and ultimately became zero at 2.1% O2 in N2 . The extent of sensitization increased with rising concentration of acetone up to the first peak (50% acetone in water) under anaerobic condition . Further increase in acetone concentration resulted a maximum response seen at 90% acetone in water . To investigate the role of hydroxyl radical in the radiation sensitization caused by acetone two different types of hydroxyl radical scavengers (t-butanol and iso-propanol) were used. Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Pneumoftiziol, 1989 Jan-Mar, 38(1), 39 - 47 {Pleural puncture biopsy--a diagnostic method in serofibrinous tuberculous pleurisy in children}; Murgoci G et al.; The biopsy of the parietal biopsy with the help of a needle has become an essential method of diagnosis in the adult patients with a pleural malady of an unknown etiology and may become a complementary paraclinical method in children . A sure diagnosis in the tubercular serofibrinous pleurisy with children is made by rendering evident the Koch bacillus on the direct test or in culture and by rendering evident the tubercular lesions on fragments of pleural biopsy puncture . The study is supported on the results obtained at the pleural biopsy puncture performed on a number of 6 children . In four cases out of six the histological test of the pleural fragment has rendered evident the presence of tubercular lymphoepithelioid nodulus with central necrosis, thus carrying the argument of certitude . The pleural biopsy puncture allows an early and sure diagnosis in over two-thirds of the pleurisies of a tubercular etiology. Free Radic Biol Med, 1989, 7(2), 165 - 70 Electron spin resonance detection of oxygen-centred radicals in murine macrophages stimulated with bacterial endotoxin; Jackson SK et al.; The production of oxygen radicals by Bacille-Calmette-Guerin primed mouse macrophages stimulated with bacterial endotoxin has been investigated . Superoxide radicals were spin-trapped in this system with dimethylpyrroline-N-oxide after a lag period of 20-40 minutes . The electron spin resonance signals due to the superoxide radical adduct could be inhibited by superoxide dismutase but not by catalase. Semin Surg Oncol, 1989, 5(4), 247 - 54 Diagnosis and management of superficial bladder cancer; Soloway MS; Superficial transitional cell carcinoma is defined as a transitional cell urothelial tumor that is confined to the mucosa, stages Ta or CIS, or with invasion of the lamina propria, T1 . The initial treatment is transurethral resection with an attempt to remove all tumor . This should provide an accurate histologic grade and stage, and from this information a prognosis can be determined . The important predictive factors that correlate with a new occurrence or true recurrence and the development of a subsequent tumor with muscle invasion are a high tumor grade, lamina propria invasion, a positive cytology following resection, multifocal tumors, dysplasia or carcinoma in situ from mucosal biopsies of normal appearing urothelium, and a prior history of bladder cancer . Based on these factors, the recurrence rate varies from 30 to 80% and progression with a muscle invasive tumor up to 30% . Intravesical chemotherapy or "immunotherapy" following tumor resection has been shown to diminish the likelihood of a recurrence . Thiotepa has been used for the longest period of time . It is relatively inexpensive, safe if myelosuppression is closely monitored, and effective . Mitomycin C was more effective than Thiotepa in randomized trials, but is significantly more expensive . This has retarded its use as a first-line agent . It has been shown to eradicate persistent tumor in 30 to 40% of patients who have failed Thiotepa . Mitomycin C is also highly effective when used for prophylaxis . Intravesical bacillus Calmette-Guerin (BCG) has recently been demonstrated to be an effective intravesical therapeutic agent . It is effective both for treatment and prophylaxis . BCG is relatively safe and inexpensive.(ABSTRACT TRUNCATED AT 250 WORDS) Anim Genet, 1989, 20(2), 167 - 78 Genetic and other effects on antibody and cell mediated immune response in swine leucocyte antigen (SLA)-defined miniature pigs; Mallard BA et al.; Miniature pigs of eight swine leucocyte antigens (SLA) haplotypes were immunized with sheep erythrocytes (SRBC), hen egg white lysozyme (HEWL) and the synthetic peptide (T, G)-A--L to induce antibody . Bacillus Calmette Geurin (BCG) and dinitrochlorobenzene (DNCB) were used to induce cell mediated immune response (CMI) . Analysis of variance by least squares was used to assess the effects of SLA haplotype, sire, dam, litter and sex of pig on the magnitude of the primary and secondary antibody response and on dermal delayed type hypersensitivity induced by purified protein derivative of tuberculin (PPD) and DNCB-induced contact hypersensitivity . The statistical model accounted for 43.50-77.30% of the observed variability in antibody and CMI at various times after immunization or challenge . While SLA had a significant effect on both antibody and CMI to some antigens at some, but not all times, sire, dam and litter were more frequently significant and to a greater degree . Haplotypes dd, dg and gg produced more antibody to SRBC and (T, G)-A--L while dg and gg had higher primary, but not secondary antibody response to HEWL . Delayed hypersensitivity to PPD was most marked in pigs of dd, dg and gg haplotypes while contact hypersensitivity to DNCB was expressed least in the dg and gg haplotype pigs . Heritability estimates were high for response to (T, G)-A--L and HEWL indicating feasibility of selective breeding for these traits. Przegl Epidemiol, 1989, 43(4), 355 - 65 {Determination of the enterotoxic properties of Bacillus cereus}; Stec E; Using indirect biological investigations and various experimental models the enterotoxic properties of 60 strains of B . cereus were determined . The determination of biochemical and enzymatic features was insufficient for indicating the enterotoxic strains and differentiating them from the non-enterotoxic ones . In the dermonecrotic test, vascular permeability test and in cultures on cells reliable criteria for determination of enterotoxicity were obtained as confirmed by the response of the ileum . The criterion of enterotoxicity determined in these tests requires raising of the index of ileum pathogenicity to 0.33. Przegl Epidemiol, 1989, 43(4), 345 - 54 {The importance of Bacillus cereus in food poisoning}; Stec E; In the light of the available literature the present knowledge on the role of Bacillus cereus in alimentary intoxications is discussed, particularly the occurrence and role of B . cereus in food pollution, the most frequent alimentary intoxications in the world are reviewed, the biological tests on animals and studies on humans are described, and the metabolites of the organism are presented . Particular attention is given to the properties of enterotoxins responsible for diarrhoea and vomiting.
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